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ISSN: 0738-8551 (print), 1549-7801 (electronic)

Crit Rev Biotechnol, Early Online: 1–7

! 2013 Informa Healthcare USA, Inc. DOI: 10.3109/07388551.2013.803020

REVIEW ARTICLE

Biotransformation and bioconversion of phenolic compounds

obtainment: an overview
Jose Valdo Madeira Junior, Camilo Barroso Teixeira and Gabriela Alves Macedo
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Food Science Department, College of Food Engineering, Campinas State University, Sao Paulo, Brazil

Abstract Keywords
Phenolic compounds have recently been recognized for their influence on human metabolism, Agro-industrial co-products, bioprocess,
acting in the prevention of some chronic diseases as well as proving to be important enzymatic reactions, phenolic compounds,
antioxidants in food. Nevertheless, the extraction and concentration processes are usually solid-state fermentation
carried out by organic solvent extraction from natural sources and can generate some
drawbacks like phenolic compound degradation, lengthy process times and low yields. As a History
solution, some eco-friendly technologies, including solid-state fermentation (SSF) or enzymatic-
assisted reaction, have been proposed as alternative processes. This article reviews the Received 5 July 2012
extraction of phenolic compounds from agro-industrial co-products by solid-state fermenta- Revised 17 April 2013
tion, even as friendly enzyme-assisted extractions. It also discusses the characteristics of each Accepted 2 May 2013
bioprocess system and the variables that affect product formation, as well as the range of Published online 15 July 2013
substrates, microorganisms and enzymes that can be useful for the production of bioactive
phenolic compounds.
For personal use only.

Introduction hydroxycinnamic acids, flavanols, flavonols, anthocyanins

and tannins (Martins et al., 2011). These compounds are
In recent years, attention has turned to phenolic compounds
considered important to the plants, both structurally and
with biological activity due to their ability to promote benefits
physiologically. In plants, they can attract pollinating insects,
for human health, such as reduction in the incidence of some
contribute to the pigmentation (flowers and fruits), act as
neurodegenerative diseases, reduction in the occurrence of
antioxidant agents, protect against UV light, and finally, play a
factors linked to cardiovascular disease and antioxidant, anti-
role in plant growth and development, preventing the action of
mutagenic, anti-allergenic, anti-inflammatory and anti-micro-
pathogens and predators (Martins et al., 2011; Soto
bial activities (Martins et al., 2011; Tripoli et al., 2007). Some
et al., 2011).
phenolic compounds may have an important technological
Currently, phenolic compounds are obtained by chemical
role in the prevention of lipid oxidation and other substances,
synthesis or extraction. Phenolic compounds are recovered
thereby increasing their shelf life (Pokorný, 2007). Research
from natural sources by solid–liquid or liquid–liquid extrac-
has been intensified in order to find plants and agricultural
tion employing organic solvents in systems of heat. However,
co-products that produce bioactive phenolic compounds.
other techniques have been used to obtain these compounds,
Agricultural co-products, as well as any plant species, have
a unique profile that will produce phenolics according to
including the use of supercritical fluids, high pressure 13
20
processes, and extraction by microwave or ultrasound. Over
specific needs and the characteristics provided by the
the past 5 years, more than 1700 articles on extraction of
environment. Thus, each co-product has a concentration of
phenolic compounds have been published, demonstrating that
a specific pronounced phenolic compound, enabling the
this type of process is already well studied. So far, most
production and extraction of a variety of bioactive compounds
studies published discuss obtaining phenolic compounds
for use (Aliakbarian et al., 2011; Harrison et al., 2012;
using chemical and physical processes. As an example,
Martins et al., 2011; Pingret et al., 2012; Saad et al., 2012;
commercial ellagic acid is obtained by chemical extraction of
Wu et al., 2012).
ellagitannin-rich materials, such as plants, using acidified
Phenolic compounds are derived mainly from the secondary
methanol. Such a process results in high cost and high
metabolites of plants from phenylalanine (Treutter, 2001).
contamination of the product and is aggressive with a low
Numerous compounds of different chemical structure are
income (Aguilera-Carbó et al., 2009; Lei et al., 2001; Wilson
grouped together, including: hydroxybenzoic acids,
& Hagerman, 1990).
Some of these processes are not feasible for the food and/or
Address for correspondence: Jose Valdo Madeira Junior, Food Science pharmaceutical industries, in both cases because of cost, use
Department, College of Food Engineering, Campinas State University,
P.O. Box 6121, 13083-862, Sao Paulo, Brazil. Tel.: +55 19 3521 2175. of organic solvents and low rate of productivity (Capote et al.,
Fax +55 19 3521 1513. E-mail: [email protected] 2007; Hayat et al., 2010; Setianto et al., 2009).
 2 J. V. Madeira Jr et al. Crit Rev Biotechnol, Early Online: 1–7

The biotransformation of bioactive compounds is also an Solid-state fermentation emerged as an attractive alterna-
interesting alternative that deserves attention, since it pre- tive for obtaining phenolic compounds. However, they are
cludes the use of toxic compounds such as organic solvents in expensive compounds using current methods of extraction
the extraction. In these processes, bioactive compounds are (Martins et al., 2011; Sepúlveda et al., 2011).
obtained from natural sources by microorganisms through The first report of the production of phenolic compounds is
their secondary metabolism or by exogenous enzymatic action from Betts et al. (1974), which describes a screening of over
(Martins et al., 2011; Puri et al., 2012). 40 microorganisms for the production of compounds with
The purpose of this article is to provide an overview of the antitumor activity. According to the results, there was a 30%
study of production and obtainment of phenolic compounds increase in the production of phenolic 9-hydroxyacronycine
by biotransformation using agro-industrial co-products. by the microorganism Cunninghamella echinulata NRRL
3655 in a stirred fermentor.
Obtaining phenolic compounds by There are patents reported for the production of polyphenol
biotransformation using microbial fermentation. Kanji (1991) reported the
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production of O-methylated phenolic compounds from the

Biotransformation can be defined as chemical transformations hydroxyl grouping by the microorganism Aspergillus repens
that are catalyzed by biological systems through their in a liquid medium. Another process for the production of
effective enzyme activity or by microorganisms through gallic acid was created using a mixed culture of Aspergillus
solid-state fermentation (SSF) (Banerjee et al., 2012; Martins foetidus and Rhizopus oryzae residues, rich in tannins
et al., 2011). In addition, this bioprocess has received great (Banerjee & Mukherjee, 2003).
attention due to the potential conversion of inexpensive agro- Table 1 presents the process used for the production of
industrial co-products, as well as plants, in a great variety of phenolic compounds by solid-state fermentation. Below, some
valuable compounds. works are cited regarding solid-state fermentation used in the
In this review, processes were shown using the technique process of obtaining phenolic compounds.
of solid-state fermentation and enzymatic reaction as alter- The choice of residue interferes deeply in obtaining the
native processes for obtaining different phenolic compounds. phenolic compound of interest. Machado et al. (2012) studied
the selection of fungal strains with potential growth and
Solid-state fermentation in phenolic obtainment
release of phenolic compounds in two coffee residues, coffee
For personal use only.

Solid-state fermentation has many advantages, such as high silverskin (CS) and spent coffee ground (SCG), by solid-state
biotechnological productivity, high concentration, product fermentation. The strains GH2 Penicillium purpurogenum,
stability and growth of microorganisms in non-water soluble Neurospora crassa and Mucor sp. 3 P showed higher growth
substrates. It also has some disadvantages, such as formation conditions in the SCG and increased the release of phenolics
of a temperature gradient throughout the fermented substrate by approximately 40%. The increasing availability of phenolic
and difficulty in controlling the pH and the amount of water. compounds varies according to the type of residue. Regarding
These problems result in reduced mobility of nutrients the silverskin, the spent coffee ground has a higher content of
derived from reduced movement of the water in the substrate. phenolic compounds, such as catechin, epicatechin, chloro-
Changes in temperature and water content in the substrate can genic acid, protocatechuic acid and ferulic acid, amongst
be caused by heating resulting from fermentation, which others. Thus, the choice of residue as the substrate for
makes it difficult to control the substrate under uniform fermentation is important for achieving different phenolic
conditions. However, there is great interest in the SSF process compounds.
among researchers and industries, particularly due to the fact Besides the residue, the microorganism interferes in the
that the process is usually cheaper with higher productivity fermentation and obtainment of phenolic compounds. Cai
than submerged fermentations (Barrios-González, 2012; et al. (2012) reported the effect of three different micro-
Singhania et al., 2009). organisms in the fermentation of oat bran as a source of

Table 1. Phenolic compounds production by microbial fermentation of agroindustrial residues.

Substrate Microorganism Incubationa Phenolic compounds Increased (%)b References

Cranberry residue Lentinus edodes 120 h/28  C Ellagic acid 37.5 Vattem et al. (2004)
Cheonggukjang Bacillus pumilus 60 h/37  C Gallic acid 233.3 Cho et al. (2009)
Pistachio hulls Phanerochaete chrysosporium 16 d/30  C Caffeic acid 12.5 Abbasi et al. (2007)
Tar bush Aspergillus niger 5 d/30  C Pyrocatechol 150 Ventura et al. (2009)
Creosote bush Aspergillus niger 4 d/30  C Gallic acid 1700 Ventura et al. (2009)
Soybeans residue Lentinus edodes 30 d/25  C Catechin 83.3 McCue et al. (2004)
Soybeans residue Kluyveromyces marxianus 72 h/30  C Gallic acid 542.9 Rashad et al. (2011)
Black soybeans Bacillus subtilis 18 h/40  C Catechin 30.5 Juan & Chou (2010)
Soybean seed Trichoderma harzianum 7 d/25  C Genistin 200 Singh et. al. (2010)
Wheat bran S. cerevisiae 48 h/32  C Ferulic acid 75 Moore et. al. (2007)
Cranberry pomace Letinus edodes 60 d/25  C Gallic acid 35.1 Zheng & Shetty (2000)
Maize kernel Thamnidium elegans 4 d/28  C Gallic acid 18.2 Salar et. al. (2012)
Lupinus angustifolious seed Bacillus subtilis 48 h/30  C Catechin 489.5 Fernandez-Orozco et al. (2008)
a
Temperature and time of incubation.
b
[(FC IC)/IC]  100; IC: initial concentration, FC: final concentration.
 DOI: 10.3109/07388551.2013.803020 Biotransformation and bioconversion of phenolic compounds 3

phenolic compounds. After the solid-state fermentation, the rice by Aspergillus oryzae for 72 h at 30 C. The results
phenolics, caffeic and ferulic acids, respectively, presented: showed that the reactor increased the concentration of
230 and 790% increase for Aspergillus oryzae var. effuses, phenolics to 330 mg g 1 as compared to the conventional
170 and 450% for Aspergillus niger, 37 and 0% increase for process, the increase of which was 270 mg g 1. This result is
Aspergillus oryzae. According to the analyses, the phenolic explained because in the first case, supplemental oxygen, heat
compounds in larger quantities are caffeic and ferulic acids, transfer and water activity were monitored in the reactor,
which are in esterified form. With hydrolysis by microorgan- which increased the fungal growth and production of the
isms, these compounds increased solubility and facilitated phenolic compounds. Probably, with circulation of humidified
their extraction, showing great advantage over other processes air, the heat produced by the fermentation was dissipated and
that use high temperatures and therefore generate high energy also facilitated the oxygen permeability into the substrate. At
costs. This difference in clearance of phenolic compounds is the same time, this maintenance of humidified air maintains
due to the metabolic activity of each microorganism. This the water percentage in the substrate during fermentation,
case is related to various types of microbial enzymes and their which increases fungal growth and, consequently, the release
Critical Reviews in Biotechnology Downloaded from informahealthcare.com by Mr. Jose Valdo Madeira Junior on 07/16/13

activities. of aglycon phenolics.

After selecting the microorganism, fermentation kinetics Several other factors also influence the production of
should be performed in order to evaluate the obtainment of phenolic compounds, such as pH. The fermentation by fungus
the product during the process. Georgetti et al. (2009) Chaetomium globosum was assessed in cottonseed and
evaluated the bioconversion of polyphenol glycosides from sugarcane bagasse, under alkaline conditions for the produc-
soybeans to form non-glycosides through solid-state fermen- tion of phenolic compounds. The alkaline stress is an
tation by Aspergillus awamori. The conversion of the important factor in the production and/or release of these
glycoside to the form of phenolic non-glycoside was compounds. According to the results, there was an increase of
accompanied by production of the enzyme b-glucosidase. 620 and 500% of gallic acid in the sugarcane bagasse and
The non-glycoside form presents a greater number of free cottonseed, respectively. The results showed a linear correl-
hydroxyl groups in regard to glycoside, thus increasing their ation between increasing pH and the amount of gallic acid.
biological activity. According to the results, the concentration Probably at a pH level between 10 and 12, the enzymes
of genistein increased by 1880% in fermented soy. b-endoglucanase, b-glucosidase and b-exoglucanase were
The production of b-glucosidase increased with fungal produced, which have higher enzymatic activity and thus
For personal use only.

growth during 48 h of incubation at 30  C, showing values their respective substrates was hydrolyzed, finally releasing
of 1000 U mL 1, leading to the conclusion that the enzyme the phenolic compounds (Ravindran et al., 2011).
was responsible for the release of phenolic compounds in the Therefore, several studies have been carried out in order to
soybean during the fermentation. After 48 h, enzyme activity optimize the production of phenolic compounds, reducing the
dropped approximately 20%. In this case, studies have costs of the process, as well as evaluating the effects that
indicated that high concentrations of genistein inhibited the influence the yield of the final product. The properties of the
activity of b-glucosidase, which in turn inhibits the hydrolysis agro-industrial co-products used, such as particle size,
of phenolic glycosides. Therefore, one should evaluate the biodegradability, water absorption and water activity, in
kinetics of the release of phenolic compounds so that no addition to their chemical composition, should be evaluated
interference inhibits its extraction. for obtainoffing a high yield phenolic compounds (Martı́nez-
Another important factor for obtaining phenolic com- Ávila et al., 2012). Most papers showed that the main factors
pounds is the addition of water to the substrate for microbial affecting the fermentation are temperature, pH, aeration,
growth. A strain of Aspergillus was subjected to fermentation water activity, microorganisms, moisture and substrate.
in grape residue to increase phenolic antioxidants. According to studies, the latter three factors have further
Fermentation kinetics were performed to evaluate the time interference in the final product, in this case, the production
when gallic acid is present in higher concentration. The of phenolic compounds (Cai et al., 2012; Georgetti et al.,
results showed that after 6 h of incubation, the concentration 2009; Machado et al., 2012; Martı́nez-Ávila et al., 2012;
of gallic acid increased 100%. After 15 h of incubation, the Martins et al., 2011; Ravindran et al., 2011).
concentration of gallic acid decreased abruptly, probably
indicating that the phenolic compounds were present during
Enzymatic processing in phenolic obtainment
the metabolism of the microorganisms. During the process, it
was noted that in solid-state fermentation, adding water to the Enzyme production is an important field in biotechnology,
substrate is extremely important, especially if the substrate having worldwide sales near five billion dollars annually, with
has hemicellulose and pectin, which can absorb more water, a growth rate of approximately 6.5 to 10%, while the number
potentially leading to an increase in microbial growth in the of patents and research papers is listed (Panke et al., 2004).
substrate. The most commonly used materials for SSF are The use of enzymes, especially in biocatalysis in agro-
those with high water absorption levels, since the moisture industrial residues, has been introduced for the hydrolysis of
content of these materials can be modified during the plant cell walls, which is complexed with polyphenols. In this
bioprocess (Martı́nez-Ávila et al., 2012). scenario, it should be concluded that enzymes can act on this
In addition to the presence of water in the substrate, the substrate in plant cells.
relative humidity also affects the product of interest. Bhanja Table 2 presents some published procedures for the
et al. (2008) compared a new solid-state reactor to the extraction of phenolic compounds by the action of microbial
conventional process, for the enrichment of the phenolic in enzymes from agro-industrial wastes. The main enzymes used
 4 J. V. Madeira Jr et al. Crit Rev Biotechnol, Early Online: 1–7

Table 2. Phenolic compounds production by microbial enzyme of agroindustrial residues.

Phenolic
Substrate Enzyme(s) Enzymatic activity Incubationa compounds Increased (%)b References
1 
Corn cobs Esterase 0.367 nkat g 4 5 h/pH 5.0/50 C Coumaric acid 1100 Topakas et al. (2004)
Xylanase 1.483 nkat g 1
1
Red dragon Pectinase 10,292 PGU mL 6 h/30  C Gallic acid 87.5 Kunnika & Pranee (2011)
pomace
Hizikin Protease 2.4 U g 1 24 h/pH 8.0/56  C Gallic acid 295.3 Siriwardhana et al. (2008)
fusiformis Carboidrase 45 U g 1
Olive B-glucosidase 3000 U mL 1 2 h/pH 4.8/50  C Hydroxytyrosol 610 Khoufi et al. (2011)
Wastewater Esterase 100 U mL 1
Grape pomace Pectinase – 2 h/pH 4.0/40  C Phenolic acids 550 Maier et al. (2008)
Cellulase –
Goldenberry Cellulase – 2 h/pH 4.3/50  C Caffeic acid 102 Ramadan et al. (2008)
Pomace oil Pectinase 300 U mL 1
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Bilberry Polygalacturonase 100 nakt g 1 Puupponen-Pimiä et al. (2008)

pomace
1
Endoglucanase 38 nakt g 2 h/pH 3.5/45  C Gallic acid 37
Xylanase 39 nakt g 1
Wheat straw Xylanase 1Ug 1 2 h/pH 5.0/40  C Coumaric acid 133.3 Tapin et al. (2006)
Esterase 20 U g 1
Rice flour a-Amilase 100 U mL 1 15 min/50  C Free phenolics 139 Bhanja et al. (2008)
b-Glucosidase 6 U mL 1
a
Time, pH and temperature of incubation.
b
[(FC IC)/IC]  100; IC: initial concentration, FC: final concentration.

in the process of obtaining phenolic compounds are reported differently to each type of substrate. As previously mentioned
below, as well as the studies performed. in this article, the effects of moisture in the reaction can
Cellulases, xylanases and ligninases are enzymes that are influence the release of phenolics. However, this condition
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capable of breaking the structure of the hemicellulose, should be evaluated together with the catalytic activity of the
cellulose and lignin of a plant cell wall. They usually have enzyme.
three important complex enzymes, endo-1,4 -b-glucanase, The pectinases are a heterogeneous group of enzymes that
cellobiohydrolase and cellobiase. These enzymes work degrade pectin. These enzymes are especially important in the
cooperatively to catalyze the hydrolysis of cellulose. The industrial sector and are used in various segments, such as
effect of these enzymes’ activity is the release of cellobiose clarification of fruit juice and wine, and product manufactur-
and glucose. Cellulases have numerous applications and ing of pectin hydrolysate extract oil from seeds and pigments.
biotechnological potential for chemicals, fuel, alcoholic Degradation of the pectin molecule occurs through a
beverages, animal feed, textile, and pulp and paper coordinated action of multiple synergistic and pectinolytic
(Gnana Soundari & Sashi, 2009; Khandeparker & Numan, enzymes, including pectin, polygalacturonase, pectate-lyase
2008; Kirby, 2005; Lin et al., 2011; Paës et al., 2012; Verma and pectin-lyase. The pectin-lyase (EC 4.2.2.10) and poly-
et al., 2011). galacturonase (EC 3.2.1.15) are of great relevance to the
Min et al. (2006) studied the conditions of dry and process of depolymerization of pectin, acting in the cleavage
humidified stems of sweet potatoes for release of ferulic acid of glycosidic bonds a-1,4, polygalacturonic acid and pectin,
primarily using commercial b-glucanase. The reaction was respectively (Alimardani-Theuil et al., 2011; Kashyap et al.,
performed with enzyme mixtures, degrading plant cell walls, 2001; Pedrolli & Carmona, 2009).
Ultraflo-L Viscozyme (Novozymes A/S) and the a-amylase Oszmianski et al. (2011) studied the action of pectinase on
(Sigma-Aldrich) in a buffer solution with pH 6.0 at 37  C apple pomace to increase the availability of phenolic com-
spinning at 12 rpm for 12 h. The results showed a concentra- pounds for the enrichment of apple juice. According to the
tion of 0.5% of Viscozyme-Ultraflo L, 50 mU of released results, pectinase increased the concentration of phenolic
a-amylase and 6 mg of ferulic acid/g of substrate humidified, compounds by 245%, especially procyanidins, flavan-3-ols
which is 3 times higher compared to the dry substrate. and flavonols. Apple peels always had tangibly higher
The ferulic acid increase in the humidified substrate must concentrations of proanthocyanidins than whole apples.
have been due to the fact that the enzymes were easily Procyanidins have been shown to bind readily to cell-wall
adsorbed into the substrate, which facilitated and enhanced polysaccharides through hydrogen-bonding and/or hydropho-
the catalytic action. bic interactions.
Moore et al. (2006) evaluated commercial enzymes and The b-glucosidase (b-D-glucoside glucohydrolase,
their action regarding the release of phenolics in wheat bran. EC 3.2.1.21) catalyzes the hydrolysis of disaccharide glyco-
The reaction was conducted in the dark with 10% humidity at sides and conjugates from the non-reducing end. The
room temperature for 72 h. The results indicated that extracts b-glucosidase enzyme has numerous applications in the
of multi-enzyme complexes (e.g. Ultraflo-L- Novozymes food and pharmaceutical industries, working in the hydrolysis
A/S) had a greater effect than the addition of various purified of cellobiose to glucose, the process of conversion of
enzymes. However, each enzyme complex responds cellulose to glucose in combination with other cellulolytic
 DOI: 10.3109/07388551.2013.803020 Biotransformation and bioconversion of phenolic compounds 5
enzymes, and the release of aroma compounds in fruit juices industrial applications, in juices, beer, cosmetics, pharma-
and wine. This enzyme is also used in the hydrolysis of ceutics and chemicals. It is primarily used in the stabilization
cyanogenic compounds present in plants for hormone of the color of wine, in the leather treatment process, and for
replacement therapy (Puri et al., 2012; van den Brink & de wastewater treatment and production of gallic acid and other
Vries, 2011). phenolics (Banerjee et al., 2001; Battestin & Macedo, 2007;
Hamza et al. (2012) studied the action of the multi- Lekha & Lonsane, 1997; Madeira et al., 2012).
enzymatic complex b-glucosidase (4600 U mL 1), esterase Chamorro et al. (2012) studied the release of phenolic
(200 U mL 1), a-amylase (92 U mL 1), xylanase (5.4 U ml 1) grape residue after its reaction to the carbohydrase enzyme
and carboxymethyl-cellulase (0.6 U mL 1) in waste water (cellulase and pectinase) and tannase. The reaction medium
from olives for the production of hydroxytyrosol. According was performed at pH 5.5 at 35  C under agitation for 24 h,
to the results, the oleuropein could be hydrolyzed and with the addition of pectinase (135 U g 1), cellulase
hydroxytyrosol obtained. The reaction probably occurred (3150 U g 1) and tannase (200 U m g 1). The results showed
during the breakdown of the glycosidic bond and resulted in that both cellulase and pectinase used alone had no change in
Critical Reviews in Biotechnology Downloaded from informahealthcare.com by Mr. Jose Valdo Madeira Junior on 07/16/13

the formation of hydroxytyrosol and elenolic acid (Figure 1). the concentration of phenolics. However, with the action of
According to the results, the highest concentration of tannase, the concentration of phenolic acids increased,
hydroxytyrosol were between pH 4 and 5 and coincided with especially gallic acid. The concentration of epigallocatechin-
the highest activity of b-glucosidase, leading to the conclu- gallate, gallocatechin and epicatechin-gallate decreased
sion that the release of phenolics is directly related to the simultaneously. Therefore, it is possible to conclude that the
aforesaid action of the enzyme. Also, it was observed that tannase hydrolyzed the ester linkages of phenolic compounds,
within 30 min of stirring, the hydroxytyrosol concentration proving that it has become an important factor in the release
tripled, and after that time these values decreased. On the of phenolic compounds in grape residue.
other hand, the static extraction was doubled after 250 min, Dueñas et al. (2007) evaluated the effect of different
compared to the values before the enzymatic extraction. Thus, enzymes in the quantification of free phenolics in lentil flour.
prolonged exposure of the phenolic compounds to O2, The residue was incubated in acetate buffer with pH 5.5 at
changed their structure and bioactive function. 37  C in four reactors, each of which received a different
Tannin acyl hydrolase, commonly referred to as tannase enzyme, a-galactosidase, viscozyme, tannase and phytase.
(EC 3.1.1.20), is an inducible enzyme produced by fungi, According to the results, both the phytase and the tannase
For personal use only.

yeasts and bacteria. This enzyme is mostly characterized by released the greatest amount of gallic acid, 1–0.8 mg g 1 lentil
its activity in the polyphenol complexes and is capable of flour. Despite the considerable increase of gallic acid, phytase
hydrolyzing ester bonds (between gallic acid and glucose) and decreased the antioxidant activity of the matrix due to the
depside linkage (between two gallic acids) substrates, such as release of phosphate groups of phytic acid chelates and other
tannic acid, epicatechin-gallate, epigallocatechin-gallate and cations in them. Phytic acid is considered to be a potent iron
chlorogenic acid, among others. This enzyme has wide chelator, which in turn prevents the formation of hydroxyl

Figure 1. Enzymatic reaction to hydroxytyrosol from oleuropein (Khoufi et al., 2011).

 6 J. V. Madeira Jr et al. Crit Rev Biotechnol, Early Online: 1–7

radicals. At the same time, there is an increase in oxidation Banerjee D, Mondal KC, Pati BR. (2001). Production and characteriza-
tion of extracellular and intracellular tannase from newly isolated
cations released in the medium that is present. However, the Aspergillus aculeatus DBF 9. J Basic Microbiol, 41, 313–18.
use of tannase for the release of phenolic antioxidants has Banerjee R, Mukherjee G. (2003). Process for the preparation of gallic
become interesting for various types of agro-industrial waste. acid by co-culture. United State of American. US7118882 (B2).
This is because most of these residues can release the Banerjee S, Singh S, Rahman LU. (2012). Biotransformation studies
using hairy root cultures – a review. Biotechnol Adv, 30, 461–8.
phenolic compounds without requiring a pre-treatment such Barrios-González J. (2012). Solid-state fermentation: physiology of solid
as the action of the pectinase or cellulase, or variation in medium, its molecular basis and applications. Process Biochem, 47,
temperature or pH. 175–85.
Briefly, the majority of published studies investigated the Battestin V, Macedo GA. (2007). Tannase production by Paecilomyces
variotii. Biores Technol, 98, 1832–7.
best conditions for enzyme activity added to the residue to Betts RE, Walters DE, Rosazza JP. (1974). Microbial transformations of
release the phenolic compounds. The main variables are the antitumor compounds. 1. Conversion of acronycine to 9-hydroxya-
enzyme concentration, reaction time, pH, and particularly the cronycine by Cunninghamella echinulata. J Med Chem, 17, 599–602.
class of enzymes used, ranging in accordance with the type of Bhanja T, Rout S, Banerjee R, Bhattacharyya BC. (2008). Studies on the
performance of a new bioreactor for improving antioxidant potential
Critical Reviews in Biotechnology Downloaded from informahealthcare.com by Mr. Jose Valdo Madeira Junior on 07/16/13

residue (Chamorro et al., 2012; Dueñas et al., 2007; Hamza of rice. LWT – Food Sci Technol, 41, 1459–65.
et al., 2012; Min et al., 2006; Moore et al., 2006). Thus, Cai S, Wang O, Wu W, et al. (2012). Comparative study of the effects of
process modeling studies should be performed in order to solid-state fermentation with three filamentous fungi on the total
increase the yield of a phenolic compound of interest, wherein phenolics content (TPC), flavonoids, and antioxidants activities of
subfractions from oats (Avena sativa L.). J Agric Food Chem, 60,
the class(es) of enzyme(s) and substrate are the major 507–13.
interferences in the final product. Capote FP, Jiménez JR, De Castro MDL. (2007). Sequential (step-by-
step) detection, identification and quantitation of extra virgin olive oil
adulteration by chemometric treatment of chromatographic profiles.
Conclusion Anal Bioanal Chem, 388, 1859–65.
The microbial and enzyme biotransformation of phenolic Chamorro S, Viveros A, Alvarez I, et al. (2012). Changes in polyphenol
and polysaccharide content of grape seed extract and grape pomace
compounds seems to be a promising way to increase the after enzymatic treatment. Food Chem, 133, 308–14.
concentration of free phenolics. These bioprocesses are clean Cho KM, Hong SY, Math RK, et al. (2009). Biotransformation of
technologies with great potential for obtaining biologically phenolics (isoflavones, flavanols and phenolic acids) during the
active compounds from natural sources. In this case, the use fermentation of cheonggukjang by Bacillus pumilus HY1. Food
Chem, 114, 413–19.
of residues is of particular interest because of its availability,
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low cost and features that allow obtaining different bioactive bioactive polyphenolic compounds of lentils by the action of
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