NT30903.

FOOD PRESERVATION and PROCESSING

W-9. Fermentation Technology

Food Spoilage

Biological Changes
 Yeast: A fungus (a plant that lacks chlorophyll) that is able to ferment sugars
and that is used for producing food products such as bread and alcohol.
 Fermentation: The conversion of carbohydrates to carbon dioxide and
alcohol by yeast or bacteria.
 THEORY: Reduce the pH of the food or produce substances which make
the environment uninhabitable by other organisms.
Food Spoilage

Chemical Changes
 Chemical reactions or changes also contribute to food deterioration.
 Enzymes play a significant role in catalyzing these reactions and can be
categorized by the substance on which they act (substrate) or their mode of
action.
 An example:
 Proteases, also called proteolytic enzymes, split proteins into smaller
compounds.
Food Spoilage

Physical Changes
 The most common physical changes occurring in foods as they spoil are
evaporation, drip loss, and separation.
 Dehydrate: To remove at least 95% of the water from foods by the use of
high temperatures.
 Freeze-dry: To remove water from food when it is in a frozen state, usually
under a vacuum.
Applications of Biopreservative
Methods

 A continuous fight against microorganisms

spoiling or making food unsafe
 Extend the shelf life of food items
What is Biopreservation ?

 can be defined as the extension of shelf life and food safety by the use of
natural or controlled microbiota and/or their antimicrobial compounds.
 the most common forms is fermentation
 The Lactic Acid Bacteria (LAB), generally considered food-grade
organisms
Factors contributing to antimicrobial
activity by Microbiota
 Organic Acids
Lactic acid, acitic acid, propionic acid
 Bacteriocins
- ribosomally synthesized, extracellularly released bioactive peptides or
peptide complexes,
- primarily active against closely related organisms,
- have a bactericidal or bacteriostatic effect,
- widespread bacteriocin is Nisin
 Diacetyl and acetaldehyde have an antimicrobial effect, imparting aroma and
flavor to cultured dairy products Reuterin by Lactobacillus reuteri
 Enzymes lacto peroxidase system, lysozyme
 CO2
 hydrogen peroxide
Fermentation - Biopreservation
 Most common forms of food biopreservation
 A process based on the growth of microorganisms in foods,
whether natural or added
Fermentation-Effects on Food
 Break down of complex compounds
 Production of organic acids and alcohols
 Synthesis of Vit-B12, Riboflavin, Vit-C precursor
 Organoleptic qualities (flavour, aroma compounds)
 Improve physicochemical properties (texture, digestibility)
 Add variety to raw food items
 Extend shelf life
 Production of antimicrobial activity
Fermented Foods

 Foods that have been subjected to the action of micro-organisms or

enzymes, in order to bring about a desirable change.
 Numerous food products owe their production and characteristics to
the fermentative activities of microorganisms.
 Fermented foods originated many thousands of years ago when
presumably micro-organism contaminated local foods.
Fermented Foods

 Micro-organisms cause changes in the foods which:

 Help to preserve the food,
 Extend shelf-life considerably over that of the raw materials from
which they are made,
 Improve aroma and flavour characteristics,
 Increase its vitamin content or its digestibility compared to the raw
materials.
Fermented Foods

 The term “biological ennoblement” has been used to describe the

nutritional benefits of fermented foods.

 Fermented foods comprise about one-third of the world wide

consumption of food and 20- 40 % (by weight) of individual diets.
Fermentation Process and Products
 The overall economics of fermentation processes
are influenced by the costs of raw materials and
consumables, utilities, labour and maintenance, fixed
charges, working capital charges, factory overheads
and operating outlay.
 Fermentation products can be broadly divided into
two categories: high volume, low value
products or low volume, high value products.
 Examples of the first category include fermentation
products, whereas many fine chemicals and
pharmaceuticals are in the latter category.
Products from Microorganisms

Primary Metabolites Secondary Metabolites

Amino Acids Antibiotics
Vitamins Pigments
Polysaccharides Toxins
Ethanol Alkaloids
Acetone and Butanol Many pharmacological compounds
Primary metabolites

 Are produced during an organism’s growth phase

Secondary metabolites

 Are not essential to cell growth or function.

Food, Beverages, Additives and
Supplements
 A wide range of fermented foods and beverages have been produced as vast
economic importance.
 Fermented dairy products as result from the activities of LAB in milk, which
modify flavour and texture, and increase long-term product stability.
 Yeasts are exploited in production of alcoholic beverages, due to their ability to
ferment sugars, derived from various plant sources, to ethanol.
 Most processes use strains of S. cerevisiae, and other strains are used as baker’s
yeast for bread dough production.
Food, Beverages, Additives and
Supplements
 Several organic acids from microbial action are employed in food manufacture and
other purposes. The first human use was for acetic acid, as vinegar, produced as a
result of the oxidation of alcoholic beverages by acetic acid bacteria.
 A further aerobic fermentation involves citric acid production by A. niger, which
has become a major industrial fermentation product, as it has numerous food and
non-food applications.
 Most of the amino acids and vitamins used as supplements in human food and
animal feed are produced most economically by microorganisms.
Health-care Products

 Antibiotics are probably the most important compounds produced by industrial

microorganisms. Most are secondary metabolites synthesized by filamentous fungi and
bacteria, particularly is actinomycetes.
 50 out of 4000 antibiotics have now been used regularly in antimicrobial chemotherapy.
The most medically useful antibiotics are the b-lactams, penicillins and
cephalosporins, aminoglycosides (e.g. streptomycin) and tetracyclines.
 Other important pharmaceutical products derived from fermentation and/or
biotransformations are alkaloids, steroids, vaccines, insulin, interferons, human growth
hormone.
Microbial enzymes

 Microbial enzymes, particularly extracellular hydrolytic enzymes, have numerous roles in

the production of a wide range of specific food and non-food products.
 Proteases, are extensively used as additives to washing powders and as microbial rennets
for the production of cheese.
 Carbohydrases are employed in the production of high-fructose corn syrup that is
produced using a-amylase and amyloglucosidase, and then isomerized to fructose
(sweeter) by a glucose isomerase.
 Immobilization of enzymes or whole cells, by their attachment to inert polymeric supports,
allows easier recovery and reuse of the biocatalyst, and some enzymes are much more
stable in this form.
Industrial chemicals and fuels

 Industrial feedstock chemicals supplied through fermentation include alcohols, solvents

such as acetone, organic acids, lipids, polysaccharides and raw materials for the
production of plastics.
 Fossil fuels, especially oil, are likely to become exhausted within the next 50–100 years,
resulting in the need to develop alternative sources of energy. Biological fuel generation
may make an increasing contribution, particularly in the conversion of renewable plant
biomass to liquid and gaseous fuels.
 Currently, methane and ethanol are the main products, although other potential fuels can
be generated using microorganisms, including hydrogen, ethane, propane and butanol.
Environmental roles of microorganisms

 Microorganisms are particularly important in wastewater treatment. Two main objectives

are to destroy all pathogenic causal water-borne diseases microbes present in the sewage.
 Second objective is to break down the organic matter in waste-water to mostly methane
and carbon dioxide, thereby producing a final effluent that can be safely discharged into
environment.
 Microbial activities can also be employed in the degradation of man-made xenobiotic
compounds within waste streams and in the bioremediation of environments contaminated
by these materials.
 Microbial-based ‘clean technology’ is also being increasingly used in the desulphurization of
fuels and the leaching of metals from low-grade mineral ores and wastes using Thiobacillus
and Sulfolobus.
Factors That Affect Final Choice of Raw
Materials

 Industrial-scale fermentations primarily use cost-effective complex substrates.

Most of C and N sources are derived from natural materials, often byproducts
of other industries.
 Cost and availability: ideally, materials should be inexpensive, and of
consistent quality and year round availability.
 Ease of handling in solid or liquid forms, along with associated transport
and storage costs.
 Sterilization requirements and any potential denaturation problems.
Factors That Affect Final Choice of Raw
Materials…..cont’d

 Formulation, mixing, complexing and viscosity characteristics that may influence

agitation, aeration and foaming during fermentation and DSP stages.
 The concentration of target product attained, its rate of formation and yield per
gram of substrate utilized.
 The levels and range of impurities, and the potential for generating further
undesired products during the process.
 Overall health and safety implications.
Cassava
 Fresh cassava contains hydrogen cyanide (HCN) or cyanic acid that should be
eliminated from any product originating from cassava to render it fit for human
consumption. Depending on the production method fermented cassava could
contains up to 20 mg / kg of HCN - against 43 mg / kg for fresh cassava.
 Gari is a fermented, gelled and dehydrated food produced from fresh cassava. It is
a popular diet in West Africa's countries. The consumption area even expands to
Central Africa.
 Polvilho is a fine tapioca/manioc/cassava flour. It can be found at latino markets in
california as "sour starch" (polvilho azedo) or "sweet starch" (polvilho doce)
Nata de Coco

 A high fiber, zero fat Philippino dessert.

 A chewy, translucent, jelly-like food product produced by the
bacterial fermentation of coconut milk.
 Commonly sweetened as a candy or dessert, and can accompany
many things including pickles, drinks, ice cream, and fruit mixes.
 Highly regarded for its high dietary fiber, and its zero fat and
cholesterol content.
 It is produced through a series of steps ranging from milk extraction,
mixing, fermentation, separating, cleaning, cutting to packaging.
Lactic Acid Bacteria-1

 Major group of Fermentative organisms.

 This group is comprised of some genera of gram-positive bacteria:

 Carnobacterium, Oenococcus, Enterococcus, Pediococcus, Lactococcus,
Streptococcus, Lactobacillus,Vagococcus, Lactosphaera,Weissells and
Lecconostoc

 Related to this group are genera such as Aerococcus, Microbacterium, and

Propionbacterium.
Lactic Acid Bacteria-2

 While this is a loosely defined group with no precise boundaries all members
share the property of producing lactic acid from hexoses.

 As fermenting organisms, they lack functional heme-linked electron transport

systems or cytochromes, they do not have a functional Krebs cycle.

 Energy is obtained by substrate-level phosphorylation while oxidising

carbohydrates.
Lactic Acid Bacteria Groups
The lactic acid bacteria can be divided into two groups based on the end products
of glucose metabolism.

 Those that produce lactic acid as the major or sole product of glucose
fermentation are designated homofermentative.

 Those that produce equal amounts of lactic acid, ethanol and CO2 are termed
heterofermentative.

 The homolactics are able to extract about twice as much energy from a given
quantity of glucose as the heterolactics.
Lactic Acid Bacteria Groups

 All members of Pediococcus, Lactococcus, Streptococcus,Vagococcus, along with

some lactobacilli are homofermenters.

 Carnobacterium, Oenococcus, Enterococcus, Lactosphaera,Weissells and Lecconostoc

and some Lactobacilli are heterofermenters

 The heterolactics are more important than the homolactics in producing flavour
and aroma components such as acetylaldehyde and diacetyl.
Lactic Acid Bacteria - Growth

 The lactic acid bacteria are mesophiles:

 they generally grow over a temperature range of about 10 to 40oC,
 an optimum between 25 and 35oC.
 Some can grow below 5 and as high as 45 oC.

 Most can grow in the pH range from 4 to 8. Though some as low as

3.2 and as high as 9.6.
Starter Cultures of LAB
 Traditionally the fermenting organisms came from the natural microflora or a
portion of the previous fermentation.

 In many cases the natural microflora is either inefficient, uncontrollable, and

unpredictable, or is destroyed during preparation of the sample prior to
fermentation (eg pasteurisation).

 A starter culture can provide particular characteristics in a more controlled

and predictable fermentation.
Starter Cultures
 Lactic starters always include bacteria that convert sugars to lactic acid, usually:
 Lactococcus lactis subsp. lactis,
 Lactococcus lactis subsp. cremoris or
 Lactococccus lactis subsp. lactis biovar diacetylactis.

 Where flavour and aroma compounds such as diacetyl are desired the lactic acid
starter will include heterofermentative organisms such as:
 Leuconostoc citrovorum or
 Leuconostoc dextranicum.
Starter Cultures

 The primary function of lactic starters is the production of lactic

acid from sugars

 Other functions of starter cultures may include the following:

 flavour, aroma, and alcohol production
 proteolytic and lipolytic activities
 inhibition of undesirable organisms
A good starter CULTURE will;

 Convert most of the sugars to lactic acid

 Increase the lactic acid concentration to 0.8 to 1.2 % (Titratable acidity)

 Drop the pH to between 4.3 to 4.5

Applications of biopreservation in
different food products
 Dairy products - Curd, yogurt , cheese
 Fermented meat products- Sausages,
 Vegetable products- Pickles, Sauerkraut, Kimchi,
 Fruit based products- Wine, cider
 Fresh sea foods
 Cereal products- beer
Other Applications
 Control of mycotoxin contamination
Ability to prevent or limit mycotoxinogenic mould growth,
The inhibition of aflatoxin accumulation by occurrence of a low-molecular-
weight metabolite produced by the LAB
 Control of postharvest diseases in fruits & vegetables
Microbial antagonists are used to control the growth of disease causal
organism, Characteristics of antagonist,
 Genetically stable.
 Effective at low concentrations.
 Ability to survive adverse environmental conditions
 Effective against a wide range of pathogens
 Amenable to production on an inexpensive growth medium.
 Amenable to a formulation with a long shelf life. Easy to dispense.
 Does not produce metabolites that are harmful to human health.
 Resistant to pesticides.
 Nonpathogenic to host commodity.
 Application of hurdle concept with biopreservation

 Chemical preservatives, physical treatments (heat), or mild heat, non-

thermal physical methods, increase the permeability of cell membranes,
Positively affects the activity of many bacteriocins
 Low intensity of preservation method is required
Health benefits
 Probiotic cultures-
- stabilizing or normalizing the gastrointestinal tract
- increase function of immune system
 Production of Vitamins & precursors
- Vit-B12, Riboflavin, Vit-C precursor
Requirements and regulatory status
for bacteriocins

In general, when selecting bacteriocin-producing strains for food applications

• The producing strain should have GRAS status
• Bacteriocin with broad spectrum of inhibition that includes pathogens,
spoilage microbs
• Thermostability
• Improved safety
• No adverse effect on quality and flavour
Requirements and regulatory status
for bacteriocins

 Microorganism should be in Generally Recognized as Safe (GRAS) status

 Purified bacteriocin must be approved as GRAS
 Distinguish bacteriocins from antibiotics
 Bacteriocin must be genetically and chemically identified, and its use and
efficacy must be shown
 Toxicological data and the fate of the molecule after ingestion are also
required
Why is this in my food?
 Preservatives and Additives are commonly used for two reasons.
 The first being the obvious; is to preserve food; keep it in its original form
and to extend its shelf life.
 The second being; to make the food seem appealing by giving it more color,
or by enhancing the taste to give it more flavour.
 There are many different types chemicals used in our food. We have all seen
them on our ingredient lists when grocery shopping, and they are often the
words we can’t pronounce.
Artificial Colours

 It doesn't take much to realise that the word 'artificial' on a food label is
equivalent to : Not good for you. However, not many people take the time to
realise how really bad it is.

 Artificial colors are a derivative of Petrochemicals and Coal tar and some are
even derived from insects that does not sound in any way appetizing, and in any
way healthy.
Artificial Colours
 There have also been many claims that artificial
food colors can cause depression, anxiety, and is
the main cause attention deficit disorder.

 Instead of artificial coloring, why not use naturally

occurring spices, that not only look great, but
smell and taste great too.

 There are hundreds of spices from around the

world that produce many wonderful colors, many
of them can be purchases in your local grocery
store. Spices like; Saffron, Turmeric, Batanin, and
Paprika are just a few of the many selections to
choose from.
Thank You for Listening