3 Biotech (2016) 6:174

DOI 10.1007/s13205-016-0485-8

REVIEW ARTICLE

Microbial enzymes: industrial progress in 21st century

Rajendra Singh1 • Manoj Kumar1 • Anshumali Mittal2 • Praveen Kumar Mehta3

Received: 8 April 2016 / Accepted: 1 August 2016 / Published online: 19 August 2016
Ó The Author(s) 2016. This article is published with open access at Springerlink.com

Abstract Biocatalytic potential of microorganisms have Introduction

been employed for centuries to produce bread, wine,
vinegar and other common products without understanding Microbes have been utilized since ancient human civi-
the biochemical basis of their ingredients. Microbial lization with first reported commercial application of yeast
enzymes have gained interest for their widespread uses in to produce alcoholic beverages from barley by the Baby-
industries and medicine owing to their stability, catalytic lonians and Sumerians as early as 6000 BC. The microbial
activity, and ease of production and optimization than plant enzymes have gained recognition globally for their wide-
and animal enzymes. The use of enzymes in various spread uses in various sectors of industries, e.g., food,
industries (e.g., food, agriculture, chemicals, and pharma- agriculture, chemicals, medicine, and energy. Enzyme
ceuticals) is increasing rapidly due to reduced processing mediated processes are rapidly gaining interest because of
time, low energy input, cost effectiveness, nontoxic and reduced process time, intake of low energy input, cost
eco-friendly characteristics. Microbial enzymes are capable effective, nontoxic and eco-friendly characteristics (Li
of degrading toxic chemical compounds of industrial and et al. 2012; Choi et al. 2015). Moreover, with the advent of
domestic wastes (phenolic compounds, nitriles, amines recombinant DNA technology and protein engineering a
etc.) either via degradation or conversion. Here in this microbe can be manipulated and cultured in large quanti-
review, we highlight and discuss current technical and ties to meet increased demand (Liu et al. 2013). Associated
scientific involvement of microorganisms in enzyme pro- driving factors that motivate the use of microbial enzymes
duction and their present status in worldwide enzyme in industrial applications are increasing demand of con-
market. sumer goods, need of cost reduction, natural resources
depletion, and environmental safety (Choi et al. 2015).
Keywords Microorganisms
Enzymes
Bioconversion
Global market for industrial enzymes was estimated about
Application
Industry $4.2 billion in 2014 and expected to develop at a com-
pound annual growth rate (CAGR) of approximately 7 %
over the period from 2015 to 2020 to reach nearly $6.2
billion ( 2015b: Industrial Enzyme Market). Enzymes are
biological molecules, proteinaceous in nature with the
exception of catalytic RNA molecules (ribozymes), and act
& Praveen Kumar Mehta as catalyst to support almost all of the chemical reactions
[email protected]
required to sustain life (Cech and Bass 1986). Enzymes are
1
Department of Biochemistry, VP Chest Institute, University highly specific; only accelerate the rate of particular reac-
of Delhi, Delhi 110007, India tion by lowering the activation energy without undergoing
2
Mill Hill Laboratory, Division of Structural Biology and any permanent change in them, and therefore, are vital
Biophysics, The Francis Crick Institute, London, UK biomolecules that support life (Fersht 1985; Piccolino
3
Department of Biotechnology and Food Engineering, 2000; Aldridge 2013). They require typically milder con-
Technion-Israel Institute of Technology, 32000 Haifa, Israel dition of temperature and pressure for catalyzing reactions,

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and are used as an alternative to hazardous chemical pol- reaction catalyzed, the enzyme commission has classified
lutant owing to their biodegradable and nontoxic nature the enzymes into six main classes, mentioned in Table 1.
(Mojsov 2011; Illanes et al. 2012; Choi et al. 2015). In Microorganisms are favored sources for industrial
addition to advantages of enzymes over conventional enzymes due to easy availability, and fast growth rate. Genetic
methods, there are some drawbacks of using enzymes in changes using recombinant DNA technology can easily be
healthcare and other industries. For many mammalian done on microbial cells for elevated enzyme production and
enzymes, 37°C and 7.4 are the optimal temperature and scientific development (Illanes et al. 2012). Production of
pH, respectively, and their activity is highly sensitive to microbial enzymes is a necessary event in the industrial sec-
any change in these parameters. Higher temperature tors, due to the high and superior performances of enzymes
([40°C), and a large deviation from the physiological pH from different microbes, which work well under a wide range
(7.4) lead to their denaturation, which limits the use of of varied physical and chemical conditions. Further, microbial
these macromolecules in non-physiological conditions. enzymes are used in the treatment of health disorders associ-
Additionally, they are susceptible to substrate or product ated with deficiency of human enzymes caused by genetic
inhibition and their products may cause allergic reactions. problems (Vellard 2003; Anbu et al. 2015). For instance,
The high cost of isolation and purification of enzymes and patients with inherited congenital sucrase-isomaltase defi-
their difficult recovery for subsequent reuse may discour- ciency are unable to digest sucrose, and therefore, sacrosidase
age their use (Johannes et al. 2006). (b-fructofuranoside fructohydrolase, EC 3.2.1.26) enzyme is
Enzymes are large macromolecules composed of poly- given orally to facilitate digestion of sucrose (Treem et al.
mers of amino acids connected by amide bonds, ranging 1999). In addition, phenylalanine ammonia lyase (EC
from kilodalton (insulin) to megadalton (ribosome) in 4.3.1.24) is used to degrade phenylalanine in genetic
molecular mass. Catalytic site of these macromolecules is phenylketonuria disorder (Sarkissian et al. 1999).
often buried deep within hydrophobic pockets, which The extensive application of microbes in different bio-
determines the specificity for their substrate. This speci- process is used to deliver a variety of products in applied
ficity of enzyme to catalyze reactions between one types of industries. Table 2 summarizes several applications of
chemical compound over the other provides the basis of its microorganisms to deliver a variety of products. The
classification and name. With the great advancement Schematic representation of industrial production of
achieved in the area of biochemistry after 1940, a large microbial enzymes has been shown in Fig. 1.
number of enzymes were isolated and characterized, and Enzymes, particularly of microbial origin, can be cul-
therefore, it was necessary to regulate the enzyme tured largely by gene manipulations, as per the need for
nomenclature. Thus, International Union of Biochemistry industrial applications. Applications of microbial enzymes
and Molecular Biology (IUBMB) in consultation with in food, pharmaceutical, textile, paper, leather, and other
International Union for Pure and Applied Chemistry industries are numerous and increasing rapidly over con-
(IUPAC) established an Enzyme Commission (EC) to be in ventional methods due to less harm to the environment,
charge of guiding the naming and systematic classification greater efficiency, and the higher quality products (Jordon
for enzymes (Liese et al. 2006). According to the type of 1929; Kamini et al. 1999; Gurung et al. 2013).

Table 1 Enzyme classes, types of reactions and corresponding enzymes

S. no. Class Reactions Enzymes

1 Oxidoreductases Transfer of hydrogen or oxygen or electrons Dehydrogenases, oxidases, oxygenases,

between molecules peroxidases
2 Transferases Transfer of groups of atoms from one molecule Fructosyltransferases, transketolases,
to another acyltransferases, transaminases
3 Hydrolases Hydrolytic cleavage of bonds Proteases, amylases, acylases, lipases,
phosphatases, cutinases
4 Lyases Non-hydrolytic cleavage by elimination or Pectate lyases, hydratases, dehydratases,
addition reactions decarboxylases, fumarase, argino
succinase
5 Isomerases Transfer of group from one position to another Isomerases, epimerases, racemases
within one molecule
6 Ligases Covalent joining of two molecules coupled with Synthetases, ligases
the hydrolysis of an energy rich bond in ATP
or similar triphosphates

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Table 2 Industrial applications of microbial enzymes

Industry Enzyme Function Microorganisms

Dairy Acid proteinase Milk coagulation Aspergillus sp.

Neutral proteinase Faster cheese ripening, debittering Bacillus subtilis, A. oryzae
Lipase Faster cheese ripening, flavor customized cheese, Aspergillus niger, A. oryzae
Lactase (b- Lactose reduced milk and whey products Escherichia coli, Kluyveromyces sp.
galactosidase)
Aminopeptidase Faster cheese ripening Lactobacillus sp.
catalase Cheese processing Aspergillus niger
Transglutaminase Protein cross linking Streptomyces sp.
Baking Amylase Flour adjustment, bread softness Aspergillus sp., Bacillus sp.
Maltogenic Enhance shelf life of breads Bacillus stearothermophilus
a-Amylase
Xylanase Dough conditioning Aspergillus niger
Lipase Dough stability and conditioning Aspergillus niger
Glucose oxidase Dough strengthening Aspergillus niger, Penicillium chrysogenum
Transglutaminase Laminated dough strength Streptoverticillium sp., streptomyces sp.
Beverage Pectinase Depectinization Aspergillus oryzae, Penicillium funiculosum
Glucose oxidase Oxygen removal from beer Aspergillus niger
Cellulase Fruit liquefaction Aspergillus niger, Trichoderma atroviride
a-Amylase Starch hydrolysis Bacillus, Aspergillus
b-Amylase Starch hydrolysis Bacillus, Streptomyces, Rhizopus
b-Glucanase Restrict haze formation Bacillus subtilis, Aspergillus spp.
protease Restrict haze formation Aspergillus niger
Pullulanase Starch saccharification Bacillus sp., Klebsiella sp.
Naringinase Debittering Aspergillus niger
limoninase Debittering Aspergillus niger, A. oryzae
Aminopeptidases Protein breakdown during mashing Lactobacillus brevis, L. plantarum
Animal feed Phytase Hydrolyze phytic acid to release phosphorous Aspergillus niger
Xylanase Enhanced digestibility of starch Aspergillus sp., Bacillus sp.
b-glucanase Digestive aid Aspergillus niger
Pulp and paper Lipase Pitch control Candida Antarctica
Protease Biofilm removal Bacillus subtilis
Amylase Deinking, drainage improvement Bacillus licheniformis
Xylanase Bleach boosting Trichoderma reesei, Thermomyces lanuginosus,
Aureobasidium pullulans
Laccase Non-chlorine bleaching, delignification Bacillus subtilis
Cellulase Deinking, drainage improvement Bacillus sp., Aspergillus niger
Polymer Lipase Polycondensation, ring-opening polymerization of Candida Antarctica
lactones, carbonates
Laccase Polymerization of bisphenol A Trametes hirsuta
Glucose oxidase Polymerization of anilines Aspergillus niger, Penicillium chrysogenum
Transglutaminase Crosslinking of protein Streptomyces mobaraensis
Tyrosinase Polymerization of lignin and chitosan Trichoderma reesei
Detergent Amylase Carbohydrate stain removal Aspergillus sp., Bacillus subtilis
Lipase Fat stain elimination Aspergillus oryzae, A. flavus,
Protease Protein stain removal Aspergillus oryzae, Bacillus subtilis
Cellulase Color clarification Aspergillus niger, Bacillus sp.
Cutinase Triglyceride removal Fusarium solani f. pisi
Mannanase Mannan spot removal Bacillus sp.

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Table 2 continued
Industry Enzyme Function Microorganisms

Leather Alkaline protease Dehairing, bating Alcaligenes faecalis

Neutral Protease Dehairing, soaking Aspergillus niger, A. flavus, Bacillus subtilis
Lipase Degreasing Aspergillus oryzae, A. flavus,
Amylase Fiber splitting Aspergillus sp., Bacillus subtilis
Cosmetics Superoxide Free radical scavenging, skin care Corynebacterium
dismutase Glutamicum, Lactobacillus plantarum
Protease Removal of dead skin Aspergillus niger, A. flavus, Bacillus subtilis
Endoglycosidase Teeth and gum tissue care Mucor hiemalis
laccase Hair dye Bacillus subtilis, Trametes versicolor
lipase Skin care Aspergillus oryzae, A. flavus
Organic Lipase Synthesis of pharmaceuticals, polymers, biodiesels, Aspergillus oryzae, A. flavus
synthesis biosurfactants
Glycosyl Synthesis of oligosaccharides Bacillus sp.
tranferase
Nitrile hydratase Synthesis of acrylamide, butyramide, nicotinamide Rhodococcus rhodochrous PA-34, Bacillus sp.
APB-6
Glucose Production of High fructose corn syrup Corynebacterium sp., streptomyces murinus
isomerase
Acyltransferase Synthesis of hydroxamic acids Bacillus sp. APB-6
Laccase Production of textile dyes, cosmetic pigments, flavor Trametes versicolor, Bacillus subtilis
agents, and pesticides
Waste Amidase Degradation of nitriles containing wastes Rhodococcus erythropolis
management Amylase Bioremediation of vegetables wastes B. licheniformis, Aspergillus sp.
Amyloglucosidase Starch hydrolysis for bioremediation Aspergillus niger
Lipase Degradation of crude oil hydrocarbons Aspergillus oryzae, Candida tropicalis
Nitrile hydratase Degradation of nitriles containing wastes Rhodococcus sp.
Protease Bioremediation of keratinic wastes Chrysosporium keratinophilum
Laccase Degradation of waste containing olefin unit, Trametes versicolor
polyurethane and phenolic compounds
Cutinase Degradation of plastics, Polycaprolactone Fusarium solani f. pisi
Manganese Degradation of phenolic compounds Phanerochaete chrysosporium, Coprinus
peroxidase cinereus
Lignin peroxidase Degradation of phenolic compounds Phanerochaete chrysosporium, Coprinus
cinereus
Oxygenase Degradation of halogenated contaminants Pseudomonas sp., Rhodococcus sp.

In this review, the attentions are given to present a Pharmaceutical and analytical industry
succinct role of all microbial enzymes involved in various
fields of technical applications, such as feed industry, food Enzymes have many significant and vital roles in the
processing and cosmetics. Furthermore, efforts are made to pharmaceutical and diagnostic industries. These are
draw a simple and clear scenario about the industrial extensively used as therapeutic drugs in health issues
structure of global enzyme market. associated with enzymatic deficiency and digestive disor-
ders, and in diagnostic procedures such as ELISA and
diabetes testing kits (Mane and Tale 2015).
The use of enzymes in industrial processes Enzyme applications in medicine are as extensive as in
industry and are growing rapidly. At present, most
Enzymes are used in industrial processes, such as baking, prominent medical uses of microbial enzymes are removal
brewing, detergents, fermented products, pharmaceuticals, of dead skin, and burns by proteolytic enzymes, and clot
textiles, leather processing. Here are a range of processes busting by fibrinolytic enzymes. Nattokinase (EC
showing how enzymes are used. 3.4.21.62), a potent fibrinolytic enzyme, is a promising

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ul-Haq et al. 2002; Zaidi et al. 2014). Chitosanase (EC

3.2.1.132) catalyze hydrolysis of chitosan to biologically
active chitosan oligosaccharides (COS), which is used as
antimicrobial, antioxidant, lowering of blood cholesterol
and high blood pressure, controlling arthritis, protective
effects against infections and improving antitumor prop-
erties (Kim and Rajapakse 2005; Ming et al. 2006; Zhang
et al. 2012; Thadathil and Velappan 2014). Applications of
microbial enzymes for different health problems are illus-
trated in Table 3 (Devlin 1986; Kaur and Sekhon 2012;
Sabu 2003; Vellard 2003; Mane and Tale 2015).
The extensive utilization of enzymes for scientific and
analytical purposes is used to estimate the concentration of
substrates and to determine the catalytic activity of
enzymes present in biological samples. Advances in the
enzyme technology have replaced or minimized the use to
harmful radioactive elements in different immunoassays,
which are used for the determination of a variety of pro-
teins and hormones (Palmer 2001).
Furthermore, enzymes are used in clinical diagnostic for
the quantitative determination of diabetes and other health
disorders, for example, glucose oxidase (EC 1.1.3.4) for
glucose; urease (EC 3.5.1.5) and glutamate dehydrogenase
(EC 1.4.1.2) for urea; lipase, carboxyl esterase, and glyc-
erol kinase for triglycerides; urate oxidase (EC 1.7.3.3) for
uric acid; creatinase (EC 3.5.3.3) and sarcosine oxidases
(EC 1.5.3.1) for creatinine (Dordick 2013; Le Roes-Hill
and Prins 2016). Cholesterol oxidase (EC 1.1.3.6) has also
been reported for useful biotechnological applications in
the detection and conversion of cholesterol. Putrescine
oxidase (EC 1.4.3.10) is used to detect biogenic amines,
Fig. 1 Industrial production of enzymes such as putrescine, a marker for food spoilage (Le Roes-
Hill and Prins 2016).
Enzymes are indispensable in nucleic acid manipulation
agent for thrombosis therapy (Sumi et al. 1987; Cho et al. for research and development in the field of genetic engi-
2010). Acid protease, dextranase (EC 2.4.1.2) and rho- neering, such as restriction endonucleases are used for site
danase (EC 2.8.1.1) may be used to treat alimentary dys- specific cleavage of DNA for molecular cloning (Newman
pepsia, tooth decay and cyanide poisoning, respectively et al. 1995) and DNA polymerases for the DNA amplifi-
(Okafor 2007). Lipases (EC 3.1.1.3) are the most fre- cation by polymerase chain reaction (PCR).
quently used enzymes in the organic synthesis and are used
in the synthesis of optically active alcohols, acids, esters, Food industry
and lactones (Cambou and Klibanov 1984; Saxena et al.
1999). Microbial lipases and polyphenol oxidases (EC United Nation Department of Economic and Social Affairs
1.10.3.2) are involved in the synthesis of (2R,3S)-3-(4- (UNDESA) estimates that world population is predicted to
methoxyphenyl) methyl glycidate (an intermediate for grow from 6.9 billion to 9.1 billion and food demand is
diltiazem) and 3, 4-dihydroxylphenyl alanine (DOPA, for expected to increase by 70 % by 2050 (http://www.un.org/
treatment of Parkinson’s disease), respectively (Faber waterforlifedecade/food_security.shtml). Quality food
1997). Tyrosinase (EC 1.14.18.1), an important oxidase supply issue can be addressed by the application of
enzyme, is involved in melanogenesis and in the produc- enzymes in the food industry. These biomolecules are
tion of L-Dihydroxy phenylalanine (L-DOPA). L-DOPA is efficiently involved in improving food production and
used as a precursor for the production of dopamine which components, such as flavor, aroma, color, texture, appear-
is a potent drug for the treatment of Parkinson’s disease ance and nutritive value (Neidleman 1984). The profound
and to control the myocardium neurogenic injury (Ikram- understanding of the role of enzymes in the food

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Table 3 Some therapeutic applications of microbial enzymes

Treatment Enzymes Microorganisms

Antitumor L-Asparaginase, L-glutaminase, L-tyrosinase, Escherichia coli, Pseudomonas acidovorans,

galactosidase Beauveria bassiana, Acinetobacter
Antiinflammatory Superoxide dismutase, Serrapeptase Lactobacillus plantarum, Nocardia sp.,
Mycobacterium sp., Corynebacterium
Glutamicum,
Anticoagulants Streptokinase, urokinase Streptococci sp., Bacillus subtilis
Antibiotic synthesis Penicillin oxidase, rifamycin B oxidase Penicillium sp.
Antioxidants Superoxide dismutases, glutathione peroxidases, Lactobacillus plantarum, Corynebacterium
catalase glutamicum
Skin ulcers Collagenase Clostridium perfringens
Detoxification Laccase, rhodanese Pseudomonas aeruginosa
Antibiotic resistance b-Lactamase Klebsiella pneumonia, Citrobacter freundii,
Serratia marcescens
Antiviral Ribonuclease, Serrapeptase Saccharomyces cerevisiae
Gout Uricase Aspergillus flavus
Digestive disorders a-Amylase, lipase Bacillus spp., Candida lipolytica, A. oryzae
Cyanide poisoning Rhodanase Sulfobacillus sibiricus

manufacturing and ingredients industry have improved the baking market. The baking enzymes industry is expected to
basic processes to provide better markets with safer and reach $695.1 million by 2019 growing at a CAGR 8.2 %
higher quality products. Furthermore, the enzymes gained from 2013 to 2019 (2014a: Baking Enzymes Market).
interest in new areas such as fat modification and sweetener Bread making is one of the most common food processing
technology (Li et al. 2012). Currently, the food and bev- techniques globally. The use of enzymes in bread manu-
erage segment dominated the industrial enzyme market and facturing shows their value in quality control and efficiency
it is projected to reach a value of $2.3 billion by 2020 of production. Amylase, alone or in combination with other
(2015c: Industrial Enzymes Market). In beverage and food enzymes, is added to the bread flour for retaining the mois-
industry, enzymes are added to control the brewing process ture more efficiently to increase softness, freshness and shelf
and produce consistent, high-quality beer; to enhance the life. Additionally, lipase and xylanase (EC 3.2.1.8) are used
functional and nutritional properties of animal and veg- for dough stability and conditioning while glucose oxidase
etables proteins by the enzymatic hydrolysis of proteins, and lipoxygenase added to improve dough strengthening and
for higher juice yield with improved color and aroma. whiteness. Transglutaminase (EC 2.3.2.13) is used in baking
The application of enzymes in food industry is seg- industry to enhance the quality of flour, the amount and
mented into different sectors, such as baking, dairy, juice texture of bread and the texture of cooked pasta (Kuraishi
production and brewing. Worldwide, microbial enzymes et al. 1997; Moore et al. 2006; Kieliszek and Misiewicz
are efficiently utilized in bakery—the principal application 2014). Lipases are also used to improve the flavor content of
market in food industry—to improve dough stability, bakery products by liberating short-chain fatty acids through
crumb softness and structure, and shelf life of products. esterification and to prolong the shelf life of the bakery
Increased uses of microbial enzymes in cheese processing products (Andreu et al. 1999; Dauter et al. 1999; Monfort
are largely responsible for the use of enzymes in dairy et al. 1999; Collar et al. 2000; Kirk et al. 2002; Fernandes
industry, which is the next largest application industry 2010; Li et al. 2012; Adrio and Demain 2014).
followed by the beverages industry.
Dairy industry
Baking industry
Dairy enzymes, an important segment of food enzyme
Baking enzymes are used for providing flour enhancement, industry, are used for the development and enhancing
dough stability, improving texture, volume and color, organoleptic characteristics (aroma, flavor and color) and
prolonging crumb softness, uniform crumb structure and higher yield of milk products. The use of enzymes (pro-
prolonging freshness of bread. To meet rising demand for teases, lipases, esterases, lactase, aminopeptidase, lyso-
quality, enzymes are seen as natural solutions in today’s zyme, lactoperoxidase, transglutaminase, catalase, etc.) in

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dairy market is well recognized and varies from coagulant by the addition of enzymes. Enzymes digest pectin, starch,
to bio-protective enzyme to enhance the shelf life and proteins and cellulose of fruits and vegetables and facilitate
safety of dairy products. Dairy enzymes are used for the improved yields, shortening of processing time and
production of cheese, yogurt and other milk products (Pai enhancing sensory characteristics (Mojsov 2012). Amy-
2003; Qureshi et al. 2015). lases are used for clarification of juices to maximize the
Rennet, a combination of chymosin and pepsin, is used production of clear or cloudy juice (Vaillant et al. 2001;
for coagulation of milk into solid curds for cheese pro- Sivaramakrishnan et al. 2006). Cellulases and pectinases
duction and liquid whey. Currently, approximately 33 % of are used to improve extraction, yield, cloud stability and
global demand of cheese produced using microbial rennet. texture in juices (Bhat 2000; Kashyap et al. 2001; Garg
Other proteases find applications for accelerated cheese et al. 2016). Naringinase (EC 3.2.1.40) and limoninase,
processing and in reduction of allergenic properties of milk debittering enzymes, hydrolyze bitter components and
products (Qureshi et al. 2015). Currently, lipases are improves the quality attributes of citrus juices (Hotchkis
involved in flavor improvement, faster cheese preparation, and Soares 2000; Li et al. 2012). Pectin, a structural
production of customized milk products, and lipolysis of heteropolysaccharide, present in nearly all fruits is required
milk fat (Sharma et al. 2001; Ghosh et al. 1996). Transg- to be maintained to regulate cloudiness of juices by poly-
lutaminase catalyzes polymerization of milk proteins and galacturonase (EC 3.2.1.15), pectin esterases (EC 3.1.1.11),
improves the functional properties of dairy products (Rossa pectin lyase (EC 4.2.2.10) and various arabanases
et al. 2011; Kieliszek and Misiewicz 2014). (Kashyap et al. 2001; Yadav et al. 2001).
Lactose intolerance is the lack of ability of human being Microbial amylases may be utilized in the distilled alco-
to digest lactose due to deficiency of lactase enzyme. holic beverages to hydrolyze starch to sugars prior to fer-
Lactase (b-galactosidase, EC 3.2.1.23) catalyzes hydrolysis mentation and to minimize or remove turbidities due to starch.
of lactose to glucose and galactose, and therefore, is used The application of enzymes to hydrolyze unmalted barley and
as a digestive aid and to enhance the solubility and other starchy adjuncts facilitate in cost reduction of beer
sweetness in milk products (Soares et al. 2012; Qureshi brewing. In brewing, development of chill-hazes in beer may
et al. 2015). It is required to minimize or removal of lactose be control by the addition of proteases (Okafor 2007).
content of milk products for lactose-intolerant people to
prevent severe tissue dehydration, diarrhea, and sometimes Feed industry
fatal consequences (Kardel et al. 1995; Pivarnik et al. 1995;
Mahoney 1997). To meet the continuously increasing worldwide demand of
milk and meat consumption, growth of feed enzymes
Beverages industry occurred steadily. The use of enzymes in animal diets
initiated in the 1980s and exploded in the 1990s. Feed
The beverage industry is divided into two major groups and enzymes are gaining importance as they can increase the
eight sub-groups. The nonalcoholic group contains soft digestibility of nutrients and higher feed utilization by
drink and syrup, packaged water, fruit juices along with tea animals (Choct 2006). The global market for feed enzymes
and coffee industry. Alcoholic group comprised distilled was estimated $899.19 million in 2014 and expected to
spirits, wine and beer (Encyclopedia of Occupational reach nearly $1.3 billion by 2020, at a CAGR of 7.3 %
health and safety). Industrial enzymes are used in breweries from 2015 to 2020 (2015a: Feed Enzyme Market).
as processing aids and to produce consistent and high- Feed enzymes may be used in animal diet formulation.
quality products. In the brewing industries, microbial For instance, these are added to degrade specific feed
enzymes are used to digest cell wall during extraction of components which are otherwise harmful or no nutritional
plant material to provide improved yield, color, and aroma value. In addition, the protein dietary value of feeds
and clearer products (Kårlund et al. 2014). available for poultry may also be enhanced by the appli-
The enzyme applications are an integrating ingredient of cation of feed enzymes (Collection of information on
the current fruit and vegetable juice industry. Enzymes are enzymes 2002). Feed enzymes mainly used for poultry are
used in fruit and vegetable juice industry as processing aids phytases, proteases, a-galactosidases, glucanases, xyla-
to increase the efficiency of operation, for instance, peel- nases, a-amylases, and polygalacturonases (Walsh et al.
ing, juicing, clarification, extraction and improve the pro- 1993; Chesson 1993; Bhat 2000; Adrio and Demain 2014).
duct quality (Law 2002). Application of cellulases, The phytase, largest enzyme segment in the feed industry,
amylases, and pectinases during fruit juice processing for is used to utilize natural phosphorous bound in phytic acid
maceration, liquefaction, and clarification, improve yield in cereal-based feed (Lei and Stahl 2000; Bhat 2000; Frias
and cost effectiveness (Kumar 2015; Garg et al. 2016). The et al. 2003). Monogastric animals are unable to digest plant
quality and stability of juices manufactured are enhanced based feeds containing high amount of cellulose and

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hemicelluloses. Xylanase and b-glucanase are added to utilization of enzymes reduce processing time, energy
their feeds as these enzymes fully degrade and digest high consumption and amount of chemicals in processing.
amount of starch (Bhat 2000; Kirk et al. 2002). Proteases Enzymes are also used to enhance deinking, and bleach in
are also used in animal feeds to overcome anti-nutritional paper and pulp industry and waste treatment by increasing
factors by degrading proteins into their constituent amino biological oxygen demand (BOD) and chemical oxygen
acids. Apart from improving the nutritional value of feed demand (COD) (Srivastava and Singh 2015). Xylanases
for better feed conversion by the animals, these feed and ligninases are used in paper and pulp industries to
enzymes are gaining importance for their role in feed cost augment the value of the pulp by removing lignin and
reduction and meat quality improvement (Lei and Stahl hemicelluloses (Maijala et al. 2008). In these industries,
2001; Adrio and Demain 2014). amylases uses include starch coating, deinking, improving
paper cleanliness and drainage improvement (Kuhad et al.
Polymer industry 2011). Lipases are employed for deinking and enhancing
pitch control while cellulases are used for deinking,
To meet the increased consumption of polymers and the improving softness and drainage improvement (Kirk and
growing concern for human health and environmental Jeffries1996). Cellulase has also been used for the devel-
safety has led to the utilization of microbial enzymes for opment of the bioprocess for recycling of used printed
synthesis of biodegradable polymer. In vitro enzyme cat- papers (Patrick 2004). The application of laccase is an
alyzed synthesis of polymer is an environmental safe pro- alternative to usage and requirement of large amount of
cess having several advantages over conventional chemical chlorine in chemical pulping process; subsequently, reduce
methods (Vroman and Tighzert 2009; Kadokawa and the waste quantity that causes ozone depletion and acidi-
Kobayashi 2010). Biopolymers are environmentally fication (Fu et al. 2005). Moreover, mannases are used for
friendly materials as these are synthesized from renewable degrading glucomannan to improve brightness in paper
carbon sources via biological processes, degrade biologi- industry (Clarke et al. 2000).
cally after use and return to the natural environment as
renewable resources, such as CO2 and biomass (Hiraishi Leather industry
and Taguchi 2009). Biopolymers, such as polyesters,
polycarbonates and polyphosphates are used in various The leather industry is more customary, and therefore,
biomedical applications, e.g., orthopedic devices, tissue discharges and waste disposed from different stages of
engineering, adhesion barriers, control drug delivery, etc. leather processing are causing severe health hazards and
(Gunatillake and Adhikari 2003; Ulery et al. 2011). environmental problems (Choudhary et al. 2004). The
The biopolymers market is growing at a CAGR of 14.5 % biodegradable enzymes are efficient alternative to improve
due to high penetration of materials in industries like medi- the quality of leather and help to shrink waste (Adrio and
cal, packaging, appliances, automotive, electronics, and Demain 2014). The initial attempt for application of
furniture and the market is expected to reach nearly $3.6 enzyme in leather industry was made for dehairing process,
billion by 2018 ( 2014b: Bioplastics & Biopolymers Market). the largest process in leather preparation which require
Increasing demand of packaging materials and environ- bulk amount of enzymes like proteases, lipases and amy-
mental safety can be addressed by the biodegradable poly- lases (Sankaran 1995; Bailey 1992; Raju et al. 1996).
mers. Lipases catalyze the polymerization of lactones, cyclic Enzymatic dehairing applications are attractive because it
diesters and cyclic carbonates to produce polyesters or can preserve the hair and contribute to fall in the organic
polycarbonates (Kobayashi 2010). Lipase catalyzed poly- load released into the effluent. Enzymatic dehairing pro-
merization is an eco-friendly technique for the preparation of cesses minimize or eliminate the dependence on harmful
useful polyesters by polycondensation as well as poly-ad- chemicals, such as sulfide, lime and amines (Green 1952;
dition reactions (Vroman and Tighzert 2009). The other Money 1996; de-Souza and Gutterres 2012).
enzymes involved in biopolymer industries are laccase (EC Enzymes are required for facilitating procedure and
1.10.3.2), peroxidase and transglutaminase for forming enhancing leather quality during different stages in leather
cross-links in biopolymers to produce materials in situ by processing, such as, curing, soaking, liming, dehairing,
means of polymerization processes (Gurung et al. 2013). bating, picking, degreasing and tanning (Mojsov 2011).
The enzymes used in leather industries are alkaline pro-
Paper and Pulp industry teases, neutral proteases, and lipases. Alkaline proteases
are used to remove non fibrillar proteins during soaking, in
With increasing awareness of sustainability issues, uses of bating to make leather soft, supple and pliable. Neutral and
microbial enzymes in paper and pulp industry have grown alkaline proteases, both are used in dehairing to reduce
steadily to reduce adverse effect on ecosystem. The water wastage (Rao et al. 1998). In addition to this, lipases

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3 Biotech (2016) 6:174 Page 9 of 15 174

are used during degreasing to remove fats (Choudhary et al. dismutase (SOD, EC 1.15.1.1) is used to arrest free radicals
2004; Sharma et al. 2001). The advantages of using and to control damage to skin caused by air and water
enzymes instead of chemicals in liming are stainless pelt, pollutions, microbes and other harmful factors. SOD and
reduced odor, low BOD and COD in effluents, and peroxidases are used in combination in sunscreen cream as
improved hair recovery (Bhatia 2003). free radical scavengers to reduce erythema (Babizhayev
2006). Proteases are used in skin creams to clean and
Textile industry smoothen the skin through peeling off dead or damaged
skin (Cho et al. 2007).
The textile industry is responsible for vast generation of Other widely used enzymes in toothpaste and mouth-
waste from desizing of fabrics, bleaching chemicals and wash are endoglycosidase and papain, which are used to
dye is one of the largest contributors to environmental whiten teeth, to remove plaque and to remove odor-causing
pollution (Ahuja et al. 2004). In such industries, enzymes deposits on teeth and gum tissue (Buckingham 1985).
are used to allow the development of environmentally Laccase, oxidases, peroxidases, and polyphenol oxidases
friendly technologies in fiber processing and strategies to are used in hair dyeing (Lang and Cotteret 2004), lipase,
improve the final product quality (Choi et al. 2015). The catalase, papain, bromelain and subtilisin in skin care
main classes of enzymes involved in cotton pre-treatment (Diehl 2008); and protein disulfide isomerase, glutathione
and finishing processes are hydrolase and oxidoreductase. sulfhydryl oxidase and transglutaminase in hair waving (Li
The group of hydrolase includes amylase, cellulase, cuti- et al. 2012). Additionally, enzymes are also used in contact
nase, protease, pectinase and lipase/esterase, which are lens cleaners to remove protein films (Alfa and Jackson
involved in the biopolishing and bioscouring of fabric, anti- 2001).
felting of wool, cotton softening, denim finishing, desizing,
wool finishing, modification of synthetic fibers, etc. (Ara- Enzymes in detergents
ujo et al. 2008; Chen et al. 2013). Oxidoreductase, other
group of enzyme, includes catalase, laccase, peroxidase, Detergents represent the largest industrial application of
and ligninase, which are involved in bio-bleaching, bleach enzymes amounting to 25–30 % of the total sales of
termination, dye decolorization, fabric, wool finishing, etc. enzymes and expected to grow faster at a CAGR of about
(Mojsov 2011). A brief detail of applications of enzymes in 11.5 % from 2015 to 2020 (2014c: Global Market for
textiles industries are shown in Table 4. Enzymes in Industrial Applications). Enzymes have con-
tributed significantly to the growth and development of
Enzymes in cosmetics industrial detergents, which is a prime application area for
enzymes today. Detergents are used in miscellaneous
The applications of enzymes in cosmetics have been con- applications as dishwashing, laundering, domestic, indus-
tinuously increased. Enzymes are used as free radical trial and institutional cleaning (Schafer et al. 2002). The
scavengers in sunscreen cream, toothpaste, mouthwashes, enzymes in detergent products are used to remove protein,
hair waving and dyeing (Li et al. 2012). The superoxide starch, oil and fats based stains and to increase the

Table 4 Uses of enzymes in textile industry

Enzyme Use Microorganisms

Amylase Desizing Bacillus sp., B. licheniformis

Cellulose Cotton softening, denim finishing Aspergillus niger, Penicillium funiculosum
Catalase Bleach termination Aspergillus sp.
Laccase Non-chlorine Bleaching, fabric dyeing Bacillus subtilis
Pectate lyase Bioscouring Bacillus sp., Pseudomonas sp.
Amylase Desizing Bacillus sp., B. licheniformis
Cellulose Cotton softening, denim finishing Aspergillus niger, Penicillium funiculosum
Protease Removal of wool fiber scales, degumming of silk Aspergillus niger, B. subtilis
Lipase Removal of size lubricants, denim finishing, Candida Antarctica
Ligninase Wool finishing Trametes versicolor, Phlebia radiata
Collagenase Wool finishing Clostridium histolyticum
Cutinase Cotton scouring, synthetic fiber modification Pseudomonas mendocina, Fusarium solani pisi, Thermomonospora fusca

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174 Page 10 of 15 3 Biotech (2016) 6:174

effectiveness of detergents (Kirk et al. 2002; Hasan et al. (Gentile et al. 1992). Oxidoreductases, such as polyphenol
2010). The enzymes in laundry detergents are weight oxidase is involved in the synthesis of 3,4-dihydrox-
efficient, cleave off damaged cotton fibers, improve ylphenyl alanine (DOPA), a chemical used in the treatment
whiteness, color and fabric care. Enzymes mainly used in of Parkinson’s disease (Faber 1997). Oligosaccharides and
detergent products are of hydrolase group and currently, polysaccharides, play vital roles in cellular recognition and
most commonly used enzymes are amylase and protease. communication processes, are synthesized industrially
Sometimes a combination of enzymes, including proteases, using high regio- and stereoselectivity of glycosyltrans-
amylases, pectinases, cellulases and lipases used to ferases (Ginsburg and Robbins 1984). Lyases are involved
increase efficiency on stain cleaning and fabric care (Li in organic synthesis of cyanohydrins from ketones, acry-
et al. 2012). lamide from acrylonitrile, malic acid from fumaric acid
Amylases and lipases are effective on removing starchy (Faber 1997; Zaks 2001). The nitrile hydratase mediated
food deposits and stains resulting from fatty products, process for the production of acrylamide is carried out by
respectively (Masse et al. 2001). Cutinase (EC 3.1.1.74), a the Nitto Chemical Company of Japan at a scale of more
hydrolytic enzyme, is used as a lipolytic enzyme in dish- than 40,000 tons per year (Zaks 2001). A multi-million ton
washing and laundry detergents (Filipsen et al. 1998; Pio of high fructose corn syrup (HFCS), an alternative sweet-
and Macedo 2009). Protease digests on organic stains, such ener to sucrose in the food and beverage industry, is pro-
as grass, blood, egg and human sweat, whereas cellulases duced every year commercially using glucose isomerase
are used to brighten colors, soften fabrics and to eliminate (Gerhartz 1990).
small fibers from the fabric without damaging the major
fibers of the fabric (Hasan et al. 2010; Kuhad et al. 2011). Waste treatment
Protease and amylase are used particularly in dishwasher
detergents to remove protein and carbohydrate containing The use of enzyme for waste management is extensive and
food particles (Keshwani et al. 2015). The application of a number of enzymes are involved in the degradation of
enzymes in detergent products is advantageous as these toxic pollutants. The industrial effluents as well as
products contain less bleaching agents, phosphates, and domestic waste contain many chemical commodities,
consequently have beneficial effects on public and envi- which are hazardous or toxic to the living being and
ronmental health (Olsen and Falholt 1998; Novozyme ecosystem. Microbial enzyme(s), alone or in combinations,
2013). are used for the treatment of industrial effluents containing
phenols, aromatic amines, nitriles, etc., by degradation or
Organic synthesis industry bioconversion of toxic chemical compound(s) to innocuous
products (Klibanov et al. 1982; Raj et al. 2006; Rubilar
Enzyme based processes for production of fine chemicals et al. 2008; Pandey et al. 2011). A number of enzymes
are rapidly gaining practical significance owing to more employed for waste treatment are amidases, amylases,
economical high purity products in an eco-environmentally amyloglucosidases, cellulases, glucoamylases, lipases,
acceptable manner (Nagasawa and Yamada 1995). nitrile hydratases, pectinases and proteases (Margesin
Enzymes are preferred in industrial chemical synthesis et al.1999; Riffaldi et al. 2006; Karigar and Rao 2011). The
over conventional methods for their high selectivity, i.e., detoxification of toxic organic compounds through oxida-
chiral, positional and functional group specific (Schmid tive coupling is mediated with oxidoreductases (Karigar
et al. 2001). Such high selectivity is extremely advanta- and Rao 2011). These enzymes, like laccase, manganese
geous in chemical synthesis as it may offer several benefits peroxidase, lignin peroxidase and tyrosinase catalyze the
such as minimal or no by-product formation, easier sepa- removal of chlorinated phenolic compounds from indus-
ration, and less environmental problems. Besides, high trial effluents (Gianfreda et al. 1999; Mai et al. 2000; Have
catalytic efficiency and mild operational conditions are and Teunissen 2001; Piontek et al. 2001; Le Roes-Hill and
advantages of enzyme mediated commercial applications. Prins 2016). The microbial enzymes are also utilized to
Catalytic potential of microorganisms have been employed recycle the waste for reuse, e.g., to recover additional oil
for hundreds of years in the production of alcohol, and from oil seeds, to convert starch to sugar, to convert whey
cheese for industrial synthetic chemistry (Johannes et al. to various useful products (Kalia et al. 2001; http://www.
2006). Among the enzymes in organic synthesis, lipases are unido.org/fileadmin/import/32068_35FoodWastes). Micro-
the most frequently used, particularly, in the formation of a bial oxygenases, such as monooxygenases and dioxyge-
wide range of optically active alcohols, acids, esters, and nases have a broad substrate range, and are active against a
lactones (Jaegera and Reetz 1998; Hasan et al. 2006). wide range of compounds, including the chlorinated
Lipases are used for the production of (S, R)-2, 3-p- aliphatics (Fetzner and Lingens 1994; Arora et al. 2009).
ethoxyphenylglycyclic acid, an intermediate for diltiazem These are used in the degradation of halogenated organic

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3 Biotech (2016) 6:174 Page 11 of 15 174

compounds containing pollutants, like herbicides, insecti- Pharmaceutical enzymes cover almost 50 % of total
cides, fungicides, hydraulic and heat transfer fluids, plas- enzyme demand in India, followed by detergent enzymes
ticizers, and intermediates for chemical synthesis (Fetzner (20 %) and textile enzymes (20 %) (Binod et al. 2013).
and Lingens 1994; Karigar and Rao 2011). In 2012, industrial enzymes market globally including
the market in India was at marginal position with a net
value of around $105 million, but expected to grow sig-
Indian enzymes market nificantly with an average of C10 % per year through 2017
to reach nearly $173 (http://www.sebi.gov.in/cms/sebi_
Around the globe, enzyme market is dominated by the food data/attachdocs/1453372309087.pdf). Greater than 60 %
and beverage products, and drug industry that go directly or enzyme market in India was contributed by the multina-
indirectly for human consumption. The biggest challenge tionals companies, whereas the rest was met by the
in front of fast growing economies such as India is to domestic manufacturers. The domestic consumption of
provide food and healthcare to even their larger population. enzymes for 2011-12 stood at about $110 million, while the
India, an agriculture-based economy, is predicted to grow exports raked $32 million in revenues during this period
at 7.9 % by 2018 (http://data.worldbank.org/country/india) (Biospectrum 2012). In 2012-13, Advanced Enzymes
and an attractive market that is opening her doors for technologies Ltd., the largest manufacturer and exporter of
industrial enzyme based manufacturing sector. Indian enzyme products in India, had nearly 30 % share in the
biotech sector accounts 2 % of the global biotech market, enzymes industry and second in line after 44 % market
but it is gaining worldwide visibility due to the investment share of Denmark based Novozymes. Other prominent
opportunities as well as its research output (Binod et al. manufacturers were Rossari Biotech, Maps Enzymes,
2013). Recently, Bharat Biotech, a Hyderabad-based Lumis Biotech and Zytex (CRISIL Research 2013)
pharma company, has developed world first Zika virus (Table 5). The demand of enzymes by their type is illus-
vaccine, which is ready for pre-clinical trials (http://www. trated in Fig. 2 (CRISIL Research 2013), (http://www.sebi.
huffingtonpost.in/2016/02/07/zika-virus_0_n_9179776. gov.in/cms/sebi_data/attachdocs/1453372309087.pdf).
html), demonstrating the ‘‘Make in India’’ efforts (http://
www.makeinindia.com/sector/biotechnology).

Table 5 Industrial enzyme, manufacturers and market share in India (2012–2013)

Industrial enzymes Manufacturer Established Indian market share Applications
(%)

Protease, xylanase, a-amylases, Novozymes 1983 (Indian 44 Household care, textiles, food and
glucoamylase, etc. India manufacturing beverages, oil and fats, baking,
unit) beverage alcohol etc.
Amylase, protease, phytase, xylanase, b- Advanced 1989 30 Food and beverages, pharma, animal
mannanase, a-galactosidase, etc. Enzymes feed, textiles, detergent, biofuel,
technologies etc.
Ltd.
Amylases, Proteases, Cellulases, Xylanase, MAPS 1975 Rest of the Market Textile, leather, feed, etc.
Glucoamylase, Pectinase, Catalase, Enzymes share along with
Lipase and Phytase, etc. Ltd. other manufacturers
Protease, amylase, cellulase, mannanase, Rossari biotech 1997 Textile, pharma, Food and
catalase, laccase, pectinases, etc. Beverages, feed, detergent,
chemical, etc.
Alkaline Pectinase, Amylase, cellulase, Lumisbiotech Textiles, Food and Beverages, feed,
Laccase, Catalase, lipase, protease, etc.
xylanase, b-glucanase, etc.
Protease, pectinase, amylase, xylanase Anthem 2007 paper, grain processing, beverages,
amyloglucosidase, catalases, etc. Cellutions textiles, baking, animal feed,
(India) Ltd pharmaceuticals, etc.
Lipase, penicillin amidase, pectinase, Aumgene 2004 Textile, pharma, Food and
Papain,lysozyme, etc. Biosciences Beverages, feed, detergent, etc.
Nattokinase, phytase, lipase amylase, Zytex India 1947 (1st Textile, food, nutraceuticals, animal
protease, cellulase, etc. Pvt. Ltd. manufacturing feed, etc.
unit in 1996)

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174 Page 12 of 15 3 Biotech (2016) 6:174

(2014) Bioplastics & biopolymers market by type (Bio PET, Bio PE,
India Enzyme Demand
200 PLA, PHA, Bio PBS, starch blends, and regenerated cellulose), by
application (packaging, bottles, fibers, agriculture, automotive,
150
($ milliom)

and others) & by geography trends & forecasts to 2018. http://

100 www.marketsandmarkets.com. http://www.marketsandmarkets.
50
com/market-reports/biopolymers-bioplastics-market-88795240.
html. Accessed on 22 Mar 2016
0 (2014) Global markets for enzymes in industrial applications 2014.
www.bccresearch.com. http://www.bccresearch.com/pressroom/
bio/global-market-industrial-enzymes-reach-nearly-$7.1-billion-
2018. Accessed on 26 Feb 2016
(2015) Feed enzymes market by type (phytase, protease, and non-
2002 2007 2012 2017 starch polysaccharides (xylanase, b-glucanase, cellulase, man-
nanase & pectinase), livestock (swine, poultry, ruminants, and
Type of Enzyme
aquatic animals) & by region—global trends & forecast to 2020.
www.bccresearch.com. http://www.marketsandmarkets.com/
Fig. 2 Industrial enzyme demand in India market-reports/feed-enzyme-market-1157.html. Accessed on 23
Mar 2016
Conclusion (2015) India industrial enzyme market: forecast and opportunities 2020.
http://www.pharmaion.com/report/india-industrial-enzymes-market-
forecast-and-opportunities-2020/10.html. Accessed 3 June 2016
The prospects of industrial uses of microbial enzymes have (2015) Industrial enzymes market by type (carbohydrases, proteases,
increased greatly in 21st century and continuously non-starch polysaccharides & others), application (food & bever-
increasing as enzymes have significant potential for many age, cleaning agents, animal feed & others), brands & by region—
global trends and forecasts to 2020. www.bccresearch.com. http://
industries to meet demand of rapidly growing population
www.marketsandmarkets.com/Market-Reports/industrial-enzymes-
and cope exhaustion of natural resources. It had been market-237327836.html. Accessed on 24 Mar 2016
showed here that the enzymes have enormous potential in Adrio JL, Demain AL (2014) Microbial enzymes: tools for biotech-
various industrial sectors that may be pharmaceuticals, nological processes. Biomolecules 4(1):117–139
Ahuja SK, Ferreira GM, Moreira AR (2004) Utilization of enzymes
food, feed, beverages, detergents, leather processing and
for environmental applications. Crit Rev Biotechnol 24:125–154
paper & pulp. Alternatively, these biomolecules may be Aldridge S (2013) Industry backs biocatalysis for greener manufac-
used consistently to meet continuously rising demand of turing. Nat Biotechnol 31:95
food supply. Enzymes of microbial origin have significant Alfa MJ, Jackson M (2001) A new hydrogen peroxide-based medical-
device detergent with germicidal properties: comparison with
potential in waste management, and consequently in the
enzymatic cleaners. Am J Infect Control 29(3):168–177
development of green environment. The enzymes are Anbu P, Gopinath SC, Chaulagain BP et al (2015) Microbial enzymes
effectively utilized in many industries for higher quality and their applications in industries and medicine 2014. Biomed
productions at accelerated rate of reaction with innocuous Res Int 2015:1–3
Andreu P, Collar C, Martı́nez-Anaya MA (1999) Thermal properties
pollution and cost effectiveness.
of doughs formulated with enzymes and starters. Eur Food Res
Technol 209:286–293
Compliance with ethical standard Araujo R, Casal M, Cavaco-Paulo A (2008) Application of enzymes
for textiles fibers processing. Biocatal Biotechnol 26:332–349
Conflict of interest The authors declare that they have no conflict of Arora PK, Kumar M, Chauhan A et al (2009) OxDBase: a database of
interest in the publication. oxygenases involved in biodegradation. BMC Res Notes 2:67
Babizhayev MA (2006) Biological activities of the natural imidazole-
Open Access This article is distributed under the terms of the containing peptidomimetics n-acetylcarnosine, carcinine and L-
Creative Commons Attribution 4.0 International License (http:// carnosine in ophthalmic and skin care products. Life Sci
creativecommons.org/licenses/by/4.0/), which permits unrestricted 78(20):2343–2357
use, distribution, and reproduction in any medium, provided you give Bailey DG (1992) Handling, greading and curing of hides and skins.
appropriate credit to the original author(s) and the source, provide a In: Pearson AM, Dutson TR (eds) Inedible meat by-products.
link to the Creative Commons license, and indicate if changes were Springer, pp 19–34. doi: 10.1007/978-94-011-7933-1_2
made. Bhat MK (2000) Cellulases and related enzymes in biotechnology.
Biotechnol Adv 18:355–383
Bhatia SC (2003) Managing industrial pollution. Macmillan India,
Delhi
References Binod P, Palkhiwala P, Gaikaiwari R et al (2013) Industrial enzymes:
present status and future perspectives for India: present scenario
(2014) Baking enzymes market by types (carbohydrase, protease, and perspectives. J Sci Ind Res 72:271–286
lipase & others), by application (bread, biscuits & cookies, cake Biospectrum 2012. http://www.biospectrumasia.com/biospectrum/
& pastry & others) & by geography—global trends & forecasts analysis/2819/indias-industrial-biotech-witnesses-growth
to 2019. www.bccresearch.com. http://www.marketsandmarkets. Buckingham KWC (1985) Methods for the treatment and prophylaxis
com/market-reports/baking-enzymes-market-250494545.html. of diaper rash and diaper dermatitis. The Procter & Gamble
Accessed on 23 Mar 2016 Company: Patent No US4556560 A

123
3 Biotech (2016) 6:174 Page 13 of 15 174

Cambou B, Klibanov AM (1984) Preparative production of optically Frias J, Doblado R, Antezana JR et al (2003) Inositol phosphate
active esters and alcohols using esterase-catalyzed stereospecific degradation by the action of phytase enzyme in legume seeds.
transesterification in organic media. J Am Chem Soc Food Chem 81(2):233–239
106:2687–2692 Fu GZ, Chan AW, Minns DE (2005) Preliminary assessment of the
Cech TR, Bass BL (1986) Biological catalysis by RNA. Annu Rev environmental benefits of enzyme bleaching for pulp and paper
Biochem 55:599–629 making. Int J Life Cycle Assess 10:136–142
Chen S, Su L, Chen J, Wu J (2013) Cutinase: characteristics, Garg G, Singh A, Kaur A et al (2016) Microbial pectinases: an
preparation, and application. Biotechnol Adv 31(8):1754–1767. ecofriendly tool of nature for industries. 3 Biotech 6(1):47–59
doi:10.1016/j.biotechadv.2013.09.005 Gentile A, Giordano C, Fuganti C et al (1992) The enzymic
Chesson A (1993) Feed enzymes. Anim Feed Sci Technol 45:65–79 preparation of (2R, 3S)-phenyl glycidic acid esters. J Org Chem
Cho SA, Cho JC, Han SH (2007) Cosmetic composition containing 57:6635–6637
enzyme and amino acid. Amorepacific Corporation (11/ Gerhartz W (ed) (1990) Enzymes in industry: production and
990):431 applications. VCH, New York
Cho YH, Song JY, Kim KM et al (2010) Production of nattokinase by Ghosh PK, Saxena RK, Gupta R et al (1996) Microbial lipases:
batch and fed-batch culture of Bacillus subtilis. New Biotechnol production and applications. Sci Prog 79:119–157
27(4):341–346. doi:10.1016/j.nbt.2010.06.003 Gianfreda L, Xu F, Bollag JM (1999) Laccases: a useful group of
Choct M (2006) Enzymes for the feed industry: past, present and oxidoreductive enzymes. Biorem J 3(1):1–25
future. Worlds Poult Sci J 62:5–15 Ginsburg V, Robbins PW (1984) Biology of carbohydrates. Wiley,
Choi JM, Han SS, Kim HS (2015) Industrial applications of enzyme New York
biocatalysis: current status and future aspect. Biotechnol Adv Green GH (1952) Unhairing by means of enzymes. J Soc Leather
33:1443–1454 Traders Chem 36:127–134
Choudhary RB, Jana AK, Jha MK (2004) Enzyme technology Gunatillake PA, Adhikari R (2003) Biodegradable synthetic polymers
applications in leather processesing. Ind J Chem Technol for tissue engineering. Eur Cell Mater 5:1–16
11:659–671 Gurung N, Ray S, Bose S, Rai V (2013) A broader view: Microbial
Clarke JH, Davidson K, Rixon JE et al (2000) A comparison of enzymes and their relevance in industries, medicine and beyond.
enzyme aided bleaching of softwood pulp using a combination Biomed Res Int: 1-18. doi:10.1155/2013/329121
of xylanase, mannanase and a-galactosidase. Appl Microbiol Hasan S, Shah AA, Hameed A (2006) Industrial applications of
Biotechnol 53:661–667 microbial lipases. Enzyme Microb Technol 39:235–251
Collar C, Martinez JC, Andreu P et al (2000) Effect of enzyme Hasan F, Shah AA, Javed S, Hameed A (2010) Enzymes used in
associations on bread dough performance: a response surface detergents: Lipases. Afr J Biotechnol 9(31):4836–4844. doi:10.
study. Food Sci Technol Int 6:217–226 5897/AJBx09.026
Collection of information on enzymes (2002) European Communities. Have RT, Teunissen PJM (2001) Oxidative mechanisms involved in
http://ec.europa.eu/environment/archives/dansub/pdfs/ lignin degradation by white-rot fungi. Chem Rev
enzymerepcomplete.pdf. Accessed 02 Mar 2016 101(11):3397–3413
CRISIL Research 2013. https://www.crisil.com/Ratings/Brochureware/ Hiraishi T, Taguchi S (2009) Enzyme-catalyzed synthesis and
News/CRISIL%20Research_ipo_grading_rationale_advanced_ degradation of biopolymers. Mini Rev Org Chem 6:44–54
enzymes_technologies.pdf Hotchkis JH, Soares NFF (2000) The use of active packaging to
Dauter Z, Dauter M, Brzozowski AM et al (1999) X-ray structure of improve citrus juice quality. International Citrus Congress (9th:
Novamyl, the five-domain ‘maltogenic’ a-amylase from Bacillus 2000: Orlando, Florida), 2003. p 1202–1205
stearothermophilus: maltose and acarbose complexes at 1.7 Å Ikram-ul-Haq, Ali S, Qadeer MA (2002) Biosynthesis of l-DOPA by
resolution. Biochemistry 38:8385–8392 Aspergillus oryzae. Bioresour Technol 85(1):25–29
De-Souza FR, Gutterres M (2012) Application of enzymes in leather Illanes A, Cauerhff A, Wilson L et al (2012) Recent trends in
processing: a comparison between chemical and coenzymatic biocatalysis engineering. Bioresour Technol 115:48–57
processes. Braz J Chem Eng 29(3):471–481 Jaegera KL, Reetz MT (1998) Microbial lipases form versatile tools
Devlin TM (1986) Textbook of biochemistry: with clinical correla- for biotechnology. Trends Biotechnol 16(9):396–403
tions ed. Wiley, New York Johannes T, Simurdiak MR, Zhao H (2006) Biocatalysis. In: Lee S
Diehl C (2008) Skin treatment preparation. Crawford Healthcare (ed) Encyclopedia of chemical processing. Taylor & Francis,
Limited: Patent No. US7468195 New York, pp 101–110
Dordick JS (2013) Biocatalysts for industry. ISBN 9781475745979 Jordon DL (1929) Red heat in salted hides. J Int Soc Leather Trade
Encyclopedia of occupational health and safety (Beverage industry). Chem 13:538–569
http://www.ilocis.org/documents/chpt65e.htm. Accessed 25 Feb Kadokawa JI, Kobayashi S (2010) Polymer synthesis by enzymatic
2016 catalysis. Curr Opin Chem Biol 14:145–153
Faber K (1997) Biotransformations in organic chemistry: a textbook. Kalia VC, Rashmi Lal S et al (2001) Using enzymes for oil recovery
Springer, Berlin from edible seeds. J Sci Ind Res 60:298–310
Fernandes P (2010) Enzymes in food processing: a condensed Kamini NR, Hemchander C, Geraldine J et al (1999) Microbial
overview on strategies for better biocatalysis. Enzym Res. enzyme technology as an alternative to conventional chemical in
doi:10.4061/2010/862537 leather industry. Curr Sci 76:101
Fersht A (1985) Enzyme structure and mechanism. W.H. Freeman, Kardel G, Furtado MM, Neto JPM (1995) Lactase na indústria de
New York laticı́nios. Revista do Instituto de Laticı́nios ‘‘Cândido Tostes’’
Fetzner S, Lingens F (1994) Bacterial dehalogenases: biochemistry, 50(294):15–17
genetics, and biotechnological applications. Microbiol Rev Karigar CS, Rao SS (2011) Role of microbial enzymes in the
58(4):641–685 bioremediation of pollutants: a review. Enzym Res. doi:10.4061/
Filipsen JAC, Appel ACM, Van Der Hidjen HTWM et al (1998) 2011/805187
Mechanism of removal of immobilized triacylglycerol by Kårlund A, Moor U, Sandell M et al (2014) The impact of harvesting,
lipolytic enzymes in a sequential laundry wash process. Enz storage and processing factors on health-promoting phytochem-
Microb Technol 23:274–280 icals in berries and fruits. Processes 2(3):596–624

123
174 Page 14 of 15 3 Biotech (2016) 6:174

Kashyap DR, Vohra PK, Chopra S (2001) Applications of pectinases Margesin R, Zimmerbauer A, Schinner F (1999) Soil lipase activity-A
in the commercial sector: a review. Bioresour Technol useful indicator of oil biodegradation. Biotechnol Tech
77:215–227 13(12):859–863
Kaur R, Sekhon BS (2012) Enzymes as drugs: an overview. J Pharm Masse L, Kennedy KJ, Chou S (2001) Testing of alkaline and
Educ Res 3(2):29–41 enzymatic pretreatment for fat particles in slaughterhouses
Keshwani A, Malhotra B, Kharkwal H (2015) Natural polymer based wastewater. Bioresour Technol 77:145–155
detergents for stain removal. World J Pharm Pharm Sci Mazzone A, Catalani M, Costanzo M et al (1990) Evaluation of
4(4):490–508 Serratia peptidase in acute or chronic inflammation of otorhi-
Kieliszek M, Misiewicz A (2014) Microbial transglutaminase and its nolaryngology pathology: a multicentre, double-blind, random-
application in the food industry. A review. Folia Microbiol ized trial versus placebo. J Int Med Res 18(5):379–388
59:241–250 Ming M, Kuroiwa T, Ichikawa S et al (2006) Production of chitosan
Kim SK, Rajapakse N (2005) Enzymatic production and biological oligosaccharides by chitosanase directly immobilized on an agar
activities of chitosan oligosaccharides (COS): a review. Carbo- gelcoated multidisk impeller. Biochem Eng J 28(3):289–294
hydr Polym 62(4):357–368 Mojsov K (2011) Applications of enzymes in the textile industry: a
Kirk TK, Jeffries TW (1996) Role of microbial enzymes for Pulp and review. In: 2nd international congress: Engineering, Ecology and
Paper Processing. ACS symposium series: Am Chem Soc Materials in the Processing Industry: Jahorina, Bosnia and
Washington, DC. pp 2–14. doi:10.1021/bk-1996-0655.ch001 Herzegovina; Tehnoloski Fakultet Zvornik, p 230–239
Kirk O, Borchert TV, Fuglsang CC (2002) Industrial enzyme Mojsov K (2012) Microbial alpha-amylases and their industrial
applications. Curr Opin Biotechnol 13:345–351 applications: a review. Int J Manage IT Eng 2(10):583–609.
Klibanov AM, Tu TM, Scott KP (1982) Enzymatic removal of ISSN 2249-0558
hazardous pollutants from industrial aqueous effluents. Enzym Money CA (1996) Unhairing and dewooling-requirement for quality
Eng 6:319–323 and the environment. J Soc Leather Tech Chem 80:175–186
Kobayashi S (2010) Lipase-catalyzed polyester synthesis-A green Monfort A, Blasco A, Sanz P et al (1999) Expression of LIP1 and
polymer chemistry. Proc Jpn Acad Ser B Phys Biol Sci LIP2 genes from Geotricum species in baker’s yeast strains and
86(4):338–365 their application to the bread-making process. J Agric Food
Kuhad RC, Gupta R, Singh A (2011) Microbial cellulases and Chem 47:803–808
their industrial applications. Enzym Res. doi:10.4061/2011/ Moore MM, Heinbockel M, Dockery P, Ulmer MH, Arendt EK
280696 (2006) Network formation in gluten-free bread with application
Kumar S (2015) Role of enzymes in fruit juice processing and its of transglutaminase. Cereal Chem 83:28–36
quality enhancement. Adv Appl Sci Res 6(6):114–124 Nagasawa T, Yamada H (1995) Microbial production of commodity
Kuraishi C, Sakamoto J, Yamazaki K et al (1997) Production of chemicals. Pure Appl Chern 67(7):1241–1256
restructured meat using microbial transglutaminase without salt Neidleman SL (1984) Applications of biocatalysis to biotechnology.
or cooking. J Food Sci 62:488–490 Biotechnol Gen Eng Rev 1:1–38
Lang G, Cotteret J (2004) Composition for the oxidation dyeing of Newman M, Strzelecka T, Dorner LF et al (1995) Structure of BamHI
keratinous fibres containing a laccase and dyeing method using endonuclease bound to DNA: partial folding and unfolding on
this composition. US2004255401 A1 DNA binding. Science 269:656–663
Law BA (2002) The nature of enzymes and their action in foods. In: Novozyme: Enzyme at work (2013) 4th edn. http://novozymes.com/
Whitehurst RJ, Law BA (eds) Enzyme in food technology. en/about-us/brochures/Documents/Enzymes_at_work.pdf
Sheffield Academic Press, Sheffileld, pp 1–18 Okafor N (2007) Biocatalysis: Immobilized enzymes and immobi-
Le Roes-Hill M, Prins A (2016) Biotechnological potential of lized cells. Modern Ind Microbiol Biotechnol, p 398. ISBN978-
oxidative enzymes from Actinobacteria. doi:10.5772/61321 1-57808-434-0 (HC)
Lei XG, Stahl CH (2000) Nutritional benefits of phytase and dietary Olsen HS, Falholt P (1998) The role of enzymes in modern
determinants of its efficacy. J Appl Anim Res 17:97–112 detergency. J Surfactants Deterg 1(4):555–567
Lei XG, Stahl CH (2001) Biotechnological development of effective Pai JS (2003) Application of microorganisms in food biotechnology.
phytases for mineral nutrition and environmental protection. Ind J Biotech 2:382–386
Appl Microbiol Biotechnol 257:474–481 Palmer T (2001) Enzymes: biochemistry, biotechnology and clinical
Li S, Yang X, Yang S et al (2012) Technology prospecting on chemistry. Horwood Publishing Ltd, Chichester, pp 345–350
enzymes: application, marketing and engineering. Comput Struct Pandey D, Singh R, Chand D (2011) An improved bioprocess for
Biotechnol J 2:1–11 synthesis of acetohydroxamic acids using DTT (dithiothreitol)
Liese A, Seelbach K, Wandrey C (2006) Industrial biotransforma- treated resting cells of Bacillus sp. APB-6. Bioresour Technol
tions, 2nd edn. Wiley-VCH, Weinheim, pp 273–447 102(11):6579–6586
Liu L, Yang H, HD Shin (2013) How to achieve high-level expression Patrick K (2004) Enzyme technology improves efficiency, cost, safety
of microbial enzymes strategies and perspectives. Bioengineered of stickies removal program. Paper Age 120:22–25
4(4):212–223 Piccolino M (2000) Biological machines: from mills to molecules.
Mahoney RR (1997) Lactose: enzymatic modification. In: Fox PF Nat Rev Mol Cell Bio 1:149–153
(ed) Advanced dairy chemistry-lactose, water, salts and vitamins Pio TF, Macedo GA (2009) Chapter 4 Cutinases: properties and
(2nd edn), Chapman & Hall, London: pp 77–125 industrial applications. Adv Appl Microbiol 66:77–95. doi:10.
Mai C, Schormann W, Milstein O et al (2000) Enhanced stability of 1016/S0065-2164(08)00804-6
laccase in the presence of phenolic compounds. Appl Microbiol Piontek K, Smith AT, Blodig W (2001) Lignin peroxidase structure
Biotechnol 54(4):510–514 and function. Biochem Soc Trans 29(2):111–116
Maijala P, Kleen M, Westin C et al (2008) Biomechanical pulping of Pivarnik LF, Senegal AG, Rand AG (1995) Hydrolytic and
softwood with enzymes and white-rot fungus Physisporinus transgalactosil activities of commercial b-galactosidase (lactase)
rivulosus. Enzym Microb Technol 43:169–177 in food processing. Adv Food Nutr Res N Y 38:1–102
Mane P, Tale V (2015) Overview of microbial therapeutic enzymes. Qureshi MA, Khare AK, Pervez A (2015) Enzymes used in dairy
Int J Curr Microbiol App Sci 4(4):17–26 industries. Int J Appl Res 1(10):523–527

123
3 Biotech (2016) 6:174 Page 15 of 15 174

Raj J, Prasad S, Bhalla TC (2006) Rhodococcus rhodochrous PA-34: Soares I, Tavora Z, Patera R et al (2012) Microorganism-produced
a potential biocatalyst for acrylamide synthesis. Process enzymes in the food industry. In: Valdez DB (ed) Food industry,
Biochem 41:1359–1363 scientific, health and social aspects of the food industry,
Raju AA, Chandrababu NK, Rose C et al (1996) Eco-friendly pp 83–94. doi:10.5772/31256
enzymatic dehairing using extracellular proteases from a bacillus Srivastava N, Singh P (2015) Degradation of toxic pollutants from
species isolate. J Am Leather Chem Assoc 91:115–119 pulp & paper mill effluent. Discovery 40(183):221–227
Rao MB, Tanksale AM, Ghatge MS et al (1998) Molecular and Sumi H et al (1987) A novel fibrinolytic enzyme (nattokinase) in the
biotechnological aspects of microbial proteases. Microbiol Mol vegetable cheese Natto; a typical and popular soybean food in
Biol Rev 62(3):597–635 the Japanese diet. Experientia 43:1110–1111
Riffaldi R, Levi-Minzi R, Cardelli R et al (2006) Soil biological Thadathil N, Velappan SP (2014) Recent developments in chitosanase
activities in monitoring the bioremediation of diesel oil- research and its biotechnological applications: a review. Food
contaminated soil. Water Air Soil Pollut 170:3–15 Chem 150:392–399. doi:10.1016/j.foodchem.2013.10.083
Rossa PN, De Sa EMF, Burin VM et al (2011) Optimization of Treem WR, McAdams L, Stanford L et al (1999) Sacrosidase therapy
microbial transglutaminase activity in ice cream using response for congenital sucrase-isomaltase deficiency. J Pediatr Gastroen-
surface methodology. Lwt Food Sci Technol 44:29–34 terol Nutr 28:137–142
Rubilar O, Diez MC, Gianfreda L (2008) Transformation of Ulery BD, Nair LS, Laurencin CT (2011) Biomedical applications of
chlorinated phenolic compounds by white rot fungi. Crit Rev biodegradable polymers. J Polym Sci B Polym Phys
Environ Sci Technol 38:227–268 49(12):832–864
Sabu A (2003) Sources, properties and applications of microbial Vaillant F, Millan A, Dornier M et al (2001) Strategy for economical
therapeutic enzymes. Indian J Biotechnol 2:234–241 optimization of the clarification of pulpy fruit juices using
Sankaran S (1995) Five decades of leather: a journey down memory crossflow microfiltration. J Food Eng 48:83–90
lane. Indian Leather, Madras Vellard M (2003) The enzyme as drug: application of enzymes as
Sarkissian CN, Shao Z, Blain F et al (1999) A different approach to pharmaceuticals. Curr Opin Biotechnol 14:444–450
treatment of phenylketonuria: phenylalanine degradation with Vroman I, Tighzert L (2009) Biodegradable Polymers. Materials
recombinant phenylalanine ammonia lyase. Proc Natl Acad Sci 2:307–344
USA 96:2339–2344 Walsh GA, Power RF, Headon DR (1993) Enzymes in the animal-
Saxena RK, Ghosh PK, Gupta R et al (1999) Microbial lipases: feed industry. Trends Biotechnol 11(10):424–430. http://www.
potential biocatalysts for the future industry. Curr Sci sebi.gov.in/cms/sebi_data/attachdocs/1453372309087.pdf
77:101–115 Yadav S, Yadav PK, Yadav D et al (2001) Pectin lyase: a review.
Schafer T, Kirk O, Borchert TV et al (2002) Enzymes for technical Process Biochem 44(1):1–10
applications. In: Fahnestock SR, Steinbüchel A (eds) Biopoly- Zaidi KU, Ali AS, Ali SA et al (2014) Microbial tyrosinases:
mers. Wiley-VCH, Cambridge, pp 377–437 promising enzymes for pharmaceutical, food bioprocessing, and
Schmid A, Dordick JS, Hauer B et al (2001) Industrial biocatalysis environmental industry. Biochem Res Int. doi:10.1155/2014/
today and tomorrow. Nature 409:258–268 854687
Sharma R, Chisti Y, Banerjee UC (2001) Production, purification, Zaks A (2001) Industrial biocatalysis. Curr Opin Chem Biol
characterization and applications of lipases. Biotechnol Adv 5(2):130–136
19:627–662 Zhang H, Sang Q, Zhang W (2012) Statistical optimization of
Sivaramakrishnan S, Gangadharan D, Nampoothiri KM et al (2006) chitosanase production by Aspergillus sp. QD-2 in submerged
a-Amylases from microbial sources—an overview on recent fermentation. Ann Microbiol 62(1):193–201
developments. Food Technol Biotechnol 44(2):173–184

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