CHAPTER-1

1

1. Introduction

Our world is changing every day both in the geological and climatic

conditions, and also similarly it is bringing the new products and services

in all sectors of the society, in terms of food habits, medicines,

nutraceuticals, cosmetics, life style products, in addition to the other

technological advances bringing the convenience or the change in tangible

and intangible usage of the daily products and services [1-3]. These

advancements and changes in the life style also brought swift changes in

the society structure and the way it operates for the needs and sustenance

of the human population and its environment. Subsequently, the products

have been changing constantly over the years to meet the changing needs

and provisions for the people and the other live stocks associated with the

people and the society. So, the growing economies and its learned societies

have been adopting the new life styles leaving the traditional lifestyle along

with the food and cultural practices [4-8].

1.1 Latest trends in the product development

The variation in products and services to the level of customized personal

needs as per the new developments in the societies all over the world, has

brought the research and development to the fore front of all the products

and services being developed and launched from time to time in to the

societies by the manufacturing and services industries and corporates. So

all the products and services offered to the society has also brought the new

management strategies and practices in the manufacturing sectors, in the

form of patents, contracts, leases, licenses etc. In addition the societies and

the regulatory bodies along with their agencies brought up the new

regulations and stringent policies on the products being released and

marketed in the society to safe guard the interests of the stake holders to

maintain and sustain the society and the environment. So every product and

CHAPTER-1

2

service provided to the society have been developed and tested by the

research and development teams in every industrial house, to meet the

requirements, standards and regulations of various agencies [9].

1.2 FMCG products and the key ingredients

Among the modern products and services, the products and services derived

from the biological sources had been growing rapidly to meet the

requirements and expectations of the society. So these biological products

and their derivatives are seen more in the form of new food products that

are baked and produced in various bakeries, hotels, malls and food

processing industries, apart from the traditional food preparations existing

among various ethnic groups of societies. To name a few in these new food

product are the most prominent Fast Moving Consumer Goods, FMCG

products like, pizzas, burgers, cola drinks, bottled water, soaps, detergents,

ice creams, fruit juices, cosmetics, meat products, fish products, cakes,

cookies etc. These FMCG products are gaining more popularity [10] due

to their unique characters like, taste, flavor, color, nutritive values and the

value additions compared to the traditional food preparations. However

these unique characters are imparted or derived from either biomolecules

or the synthetic high valued chemicals added to the products in their

formulations and processing.

1.3 Biomolecules and synthetic molecules

The biomolecules imparting the unique characters and value additions are

mostly, the enzymes of functional values, proteins of nutritional value, dyes

imparting color, terpenoids and flavonoids imparting flavor, peptides of

special function; hormones, steroids, antibiotics etc. providing the

medicinal values and so on [11]. On the other hand, the synthetic molecules

used in the preparation, processing and packing are mostly the complex

CHAPTER-1

3

molecules like, synthetic dyes, polyethylene, polyesters, phenolic

derivatives, polypropylenes, diauxins and polychlorinated biphenyls. But

these synthetic molecules are hard to digest in the biosystems of the

environment and some of them take hundreds and millions of years to get

degraded as they may be the xenobiotic compounds that are foreign to the

environment and biosystems and hence create the environmental problems

in terms of pollution and its related complications in the biota [12-15]. On

the contrary, biomolecules are derived from the biosystems and hence they

are readily accepted by the biosystems and the environment and

subsequently degraded easily without causing any pollution and its related

problems [16-20].

1.4 Advantages of biomolecules

The biomolecules being environment friendly are easily accepted by the

biosystems and hence they are in high demand for many of the industrial

applications and the personal day to day applications. Though the

biomolecules are derived mostly from either microbial systems or plant

systems or the animal systems or from any of their derivatives, the greater

emphasis was laid on the products derived from microbial sources due to

their availability, amenability and variety [21-24] for numerous

applications. Conversely, the other plant and animal derived biomolecules

attract relatively many ethical and social problems, in addition to their

lesser adaptability due to their associated technical difficulties and the low

feasibility of process development and the economics.

1.5 Microbial organisms and the derived products

Among the life forms on the earth, microbes are consider to be the

ubiquitous custodians of the Earth, as the microorganisms are estimated to

constitute about half the biomass on the Earth. Hence the microbial

CHAPTER-1

4

organisms are constantly explored from every ecological niche on the Earth

including the extreme places like the tropical forests, underground mines

and oil fields, Arctic and Antarctic ice caps, deserts, hills and mountains,

caves of extreme conditions, the dead sea, underwater hydrothermal vents,

hot springs, stratosphere [25-27] etc. in search of the novel products, as

microorganisms represent the richest repertoire of biomolecules and diverse

chemicals in nature. Some of these microbial derived biomolecules and the

high valued chemicals can be seen as the constituents in food products like

bread, cheese, yogurt, mushrooms, wine, beer, soy sauce, cake etc.; food

additives like amino acids, gelling agents, vitamins etc.; in solvents like

ethanol and butanol; in biofuels like ethanol, methanol, hydrogen, methane

etc.; and in fine chemicals like antibiotics, antifungals, hormones, enzymes,

proteins and therapeutic drugs [28-36]. Part of these biomolecules derived

from the microorganisms were listed in the Table-1 and 2 whereas the

microorganisms used as part of the edible foods [37] are listed in Table -3.

Table-1. The recent developments in the microbial biomolecules [37]

Sl.

No.

Microbial derived product and the source

microorganism

1 Natamycin derived from Streptomyces

natalensis and Streptomyces chattanoogensis

2 Bakers yeast protein derived from Saccharomyces cerevisiae

3 Yeast-malt sprout extract, derived from Saccharomyces

cerevisiae, Saccharomyces fragilis,Candida utilis

4

Carrageenan, a hydrocolloid extracted from the members of the

families Gigartinaceae and Soliericeae of the

class Rodophyceae (red seaweed) such as Chondrus crispus,

Chondrus ocellatus, Eucheuma cottonii, Eucheuma

spinosum, Gigartina acicularis, Gigartina pistillata, Gigartina

radula, Gigartina stellate.

5 Furcelleran, the refined hydrocolloid extracted from Furcellaria

fastigiata of the class Rodophyceae (red seaweed)

6 Xanthan Gum derived from Xanthomonas campestris

7 Gibberellic acid derived by fermentation from Fusarium

moniliforme

CHAPTER-1

5

Sl.

No.

Microbial derived product and the source

microorganism

8 Dried yeasts, Saccharomyces cerevisiae, Saccharomyces fragilis,

and dried torula yeast, Candida utilis

9 Bakers yeast glycan from Saccharomyces cerevisiae

10 Amyloglucosidase derived from Rhizopus niveus for use in

degrading gelatinized starch into constituent sugars

11 Carbohydrase and cellulase derived from Aspergillus niger for use

in clam and shrimp processing

12 Carbohydrase derived from Rhizopus oryzae for use in the

production of dextrose from starch

13 Catalase derived from Micrococcus lysodeikticus for use in the

manufacture of cheese

14 Esterase-lipase derived from Mucor miehei var.

Cooney et Emerson as a flavor enhancer in cheeses, fats and oils,

and milk products

15 Alpha-galactosidase derived from Morteirella

vinaceae var. raffinoseutilizer for use in the production of sucrose

from sugar beets

16

Milk-clotting enzymes, microbial for use in the production of

cheese (Milk-clotting enzymes are derived from Endothia

parasitica, Bacillus cereus, Mucor pusillus Lindt and Mucor

mieheiand Aspergillus oryzae modified to contain the gene for

aspartic proteinase fromRhizomucor miehei var Cooney et

Emerson

17 Candida guilliermondii as the organism for fermentation

production of citric acid

18 Candida lipolytica for fermentation production of citric acid.

19 A solvent extraction process for recovery of citric acid

from Aspergillus niger fermentation liquor

Table-2. Variety of biomolecules produced and derived from

microorganisms for many food and high valued chemical industries [37]

Sl.

No.

Variety of biomolecules produced and derived from the

microorganisms

1 Acetic acid may be produced by fermentation of many

microorganisms

2 Alginic acid made from certain brown algae

3 Alpha-amylase enzyme preparation from Bacillus

stearothermophilus used to hydrolyze edible starch to produce

maltodextrin and nutritive carbohydrate sweeteners.

CHAPTER-1

6

Sl.

No.

Variety of biomolecules produced and derived from the

microorganisms

4

Mixed carbohydrase and protease enzyme product derived

from Bacillus licheniformis for use in hydrolyzing proteins and

carbohydrates in the preparation of alcoholic beverages, candy,

nutritive sweeteners and protein hydrolysates.

5 Lactic acid may be produced by fermentation

Propionic acid from bacterial fermentation

7 Agar-agar, extracted from a number of related species of red algae

class Rhodophyceae

8

Brown algae, to be used dried as a flavor enhancer, are seaweeds

of the species: Analipus japonicus, Eisenia bicyclis, Hizikia

fusiforme, Kjellmaniella gyrata, Laminaria angustata,Laminaria

longirruris, Laminaria Longissima, Laminaria

ochotensis, Laminaria claustonia,Laminaria

saccharina, Laminaria digitata, Laminaria japonica, Macrocystis

pyrifera, Petalonia fascia, Scytosiphon lome

9

Red algae, to be used dried as a flavor enhancer, are seaweeds of

the species: Gloiopeltis furcata, Porphyra crispata, Porhyra

deutata, Porhyra perforata, Porhyra suborbiculata,Porphyra

tenera, Rhodymenis palmata

10 Ammonium alginate from certain brown algae

11 Calcium alginate from certain brown algae

12

Glucono delta-lactone, by oxidation of D-glucose by

microorganisms that are nonpathogenic and nontoxicogenic to

man or other animals. These include but are not restricted

toAspergillus niger and Acetobactor suboxydans

13

Insoluble glucose isomerase enzyme preparations are derived from

recognized species of precisely classified, nonpathogenic, and

nontoxicogenic microorganisms, includingStreptomyces

rubiginosus, Actinoplane missouriensis, Streptomyces

olivaceus,Streptomyces olivochromogenes, and Bacillus

coagulans grown in a pure culture fermentation that produces no

antibiotic

14 Lactase enzyme preparation from Candida pseudotropicalis for

use in hydrolyzing lactose to glucose and galactose

15 Lactase enzyme preparation from Kluyveromyces

lactis (previously called Saccharomyces lactis)for use in

hydrolyzing lactose in milk

16 Lipase enzyme preparation from Rhizopus niveus used in the

interesterfication of fats and oils.

17 Nisin preparation from Lactococcus lactis Lancefield Group N for

use as an antimicrobial agent to inhibit the outgrowth

CHAPTER-1

7

Sl.

No.

Variety of biomolecules produced and derived from the

microorganisms

of Clostridium botulinum spores and toxin formation in

pasteurized cheese spreads.

18 Potassium alginate, the potassium salt of alginic acid, derived from

certain brown algae

19

Rennet (animal derived) and chymosin preparation

from Escherichia coli K-12,Kluyveromyces

marxianus var. lactis or Aspergillus niger var. awamori to

coagulate milk in cheeses and other dairy products

20 Riboflavin biosynthesized by Eremothecium ashbyii

21 Sodium alginate, the sodium salt of alginic acid, derived from

certain brown algae

22

Butter starter distillate from milk cultures of Streptococcus

lactis, Streptococcus cremoris.Streptococcus

lactis subspecies diacetylactis, Leuconostoc

citovorum, Leuconostoc dextranicum

23 Urease enzyme preparation from Lactobacillus fermentum for use

in the production of wine

24 Vitamin B12 from Streptomyces griseus

25 Vitamin D, produced by ultraviolet irradiation of ergosterol

isolated from yeast and related fungi

26 Bakers Yeast extract from Saccharomyces cerevisiae

27 Aminopeptidase enzyme preparation from Lactococcus lactis used

as an optional ingredient for flavor development in the

manufacture of cheddar cheese.

Table-3. Food products containing microorganisms or their derivaties for

human consumption [37]

Sl.

No.

Food products containing microorganisms or their

derivatives consumed by human beings

1 Acidified milk, with or without the addition of characterizing

microbial organisms, and aroma - and flavor - producing

microbial culture.

2 Yogurt made by the lactic acid producing bacteria Lactobacillus

bulgaricus and Streptococcus thermophilus

3 Blue cheese, characterized by the presence of the

mold Penicillium roquefortii

4 Cheddar cheese, subjected to the action of a lactic acid producing

bacterial culture and clotting enzymes of animal, plant or

microbial origin used in curing or flavor development

CHAPTER-1

8

Sl.

No.

Food products containing microorganisms or their

derivatives consumed by human beings

5 Bread, rolls, and buns may contain as optional ingredients lactic-

acid producing bacteria

6 Flour may contain alpha-amylase obtained from the

fungus Aspergillus oryzae

Among the biomolecules derived from the microbial organisms, enzymes

are one of the best class of biomolecules explored and produced for various

applications. However such enzymes were also explored and be produced

from other forms of life such as plants and animals or their derivatives, but

the feasibility of commercial production of these high valued enzymes

would be less probable due to the ethical, environmental and technical

difficulties associated with the process development and the acceptability

of the final products. Conversely, the microbial derived enzymes have been

produced continuously at commercial scale and hence the process

feasibility is very high, both in terms of economic and technical feasibility

[21].

1.6 Enzymes as the revolutionary biomolecules

Enzymes are the biomolecules and they are popularly known as biocatalysts

that can accelerate the rate of reaction by several times under the normal

physiological conditions of life systems. These enzymes are the key for all

the biochemical reactions that happen in and around the biological systems.

This peculiar property of the enzymes to catalyze the reactions under the

ambient and ecofriendly conditions has put behind the harsh reaction

conditions and high temperatures and pressures that were practiced from

the centuries, and at the same time brought the convenience and safety to

the fore front at an affordable cost for many of the industries. Further these

enzymes also increased the product quality and specificity to the highest

CHAPTER-1

9

precision and accuracy and thus enhanced the product value to the

maximum levels with the minimal process and downstream operations. So,

these enzyme biomolecules revolutionized the process development and

brought the new trends in the manufacturing and processing sectors of

many chemical and process industries [35-36].

1.7 Applications of enzymes

Use of enzymes in the history of mankind was practiced since the evolution

of human civilization, to accomplish certain reactions in the industrial

applications apart from their cellular reactions, even before the nature and

functions of enzymes were understood. The ancient use of enzymes were

seen particularly in brewing of barley malt for starch conversion, and in

bating of hides for leather making. Later the crude enzyme preparations

were made from animal tissues like pancreas and stomach mucosa, or from

plant tissues like malt, jack beans and papaya tissues and such enzymes

were used in the industries of textile, leather, brewing etc. The encouraging

and established results of these crude enzyme preparations in the initial

development stages led to the search for better, less expensive and more

freely available sources of such enzymes. In that endeavor some microbial

organisms were found to produce amylases and proteases that were similar

in action to the amylases of malt and proteases of pancreas; which led

eventually to the research and development of processes for the production

of such enzymes on commercial scale [21]. In this concern, Dr. Jokichi

Takamine (1897) introduced the fungal enzymes and Biodin and Effront

(1917) introduced the bacterial enzymes to the industries as cultivated

enzymes. Since then the industrial production and utilization of enzymes

have been evolving the novel products through innovation, creativity,

affordability with value added services to the mankind [38-41]. Some of

those developments in the enzymes and their microbial sources used in

CHAPTER-1

10

laboratories and manufacturing units for the preparation of either crude or

isolated enzymes were listed in the Table-4.

Table 4. Industrial applications of enzymes and their sources [21]

Industry Application Enzyme Source* Extent

of Use+

Baking and

milling

Bread baking Amylase Fungal, malt 2

Protease Fungal 1

Beer

Mashing Amylase Malt,

bacterial 1

Chill

proofing Protease

Papin,

bromelain,

pepsin,

fungal,

bacterial

1

Carbonated

beverages

Oxygen

removal

Glucose

oxidase Fungal 3

Oxygen

removal

Glucose

oxidase Fungal 3

Cereals

Precooked

bay foods Amylase Malt, Fungal

2

Breakfast

foods Amylase Malt, Fungal

2

Condiments Protease

Papin,

bromelain,

pepsin,

fungal,

bacterial 2

Chocolate

and Cocoa Syrups Amylase

Fungal,

bacterial 2

Coffee

Coffee bean

fermentation Pectinase Fungal

2

Coffee

concentrates

Pectinase,

hemicellulase Fungal

2

Confection

ery and

Candy

Soft center

candies and

fondanta

Invertase Yeast

2

Sugar

recovery

from scrap

candy

Amylase Bacterial,

fungal

3

CHAPTER-1

11

Industry Application Enzyme Source* Extent

of Use+

Dairy

Cheese

production Rennin Animal

1

Milk,

sterilization

with

peroxide

Catalase Liver,

bacterial

3

Milk,

prevention of

oxidation

flavor

Protease Pancreatin

2

Milk, protein

hydrolyzates Protease

Papin,

bromelain,

pancreatin,

fungal,

bacterial 2

Evaporated

milk,

stabilization

Protease

Pancreatin,

pepsin,

bromelain,

fungal 4

Whole milk

concentrates Lactase Yeast

3

Ice cream

and frozen

desserts

Lactase Yeast

3

Whey

concentrates Lactase Yeast

2

Dried milk,

oxygen

removal

Glucose

oxidase Fungal

3

Distilled

beverages Mashing Amylase

Malt, fungal,

bacterial 1

Dry

cleaning,

laundry

Spot removal

Protease,

lipase,

amylase

Bacterial,

pancreatin,

fungal 1

Eggs, dried

Glucose

removal

Glucose

oxidase Fungal

1

Mayonnaise,

oxygen

removal

Glucose

oxidase Fungal

4

Feeds,

animal

Pig starter

rations

Protease,

amylase

Pepsin,

pancreatin,

bromelain,

fungal 3

CHAPTER-1

12

Industry Application Enzyme Source* Extent

of Use+

Flavors

Removal of

starch,

clarification

Amylase Fungal

3

Oxygen

removal

Glucose

oxidase Fungal

3

Fruits and

Fruit juices

Clarification,

filtration,

concentration

Pectinases Fungal

1

Low

methoxyl

pectin

Pectinesterase Fungal,

vegetable 2

Starch

removal from

pectin

Amylase Fungal

2

Oxygen

removal

Glucose

oxidase Fungal

4

Leather

Bating Protease

Bacterial,

pancreatin,

fungal 1

Unhairing Protease,

mucolytic

Bacterial,

fungal,

pancreatin 4

Meat, fish

Meat

tenderizing Protease

Papain,

bromelain,

fungal 2

Tenderizing

casings Protease

Papain,

bromelain,

fungal 3

Condensed

fish solubles Protease

Papain,

bromelain,

bacterial 2

Paper

Starch

modification

for paper

coating

Amylase Bacterial,

malt

2

Starch and

syrup

Corn syrup Amylase,

dextrinase Fungal

1

Production of

glucose

Amylase,

amyloglucosi

dase

Fungal

3

Pharmaceut

ical and

Clinical

Cold

swelling

laundry

starch

Amylase Bacterial

2

CHAPTER-1

13

Industry Application Enzyme Source* Extent

of Use+

Digestive

aids Amylase

Fungal,

pancreatin 1

Protease

Papain,

pancreatin,

bromelain,

pepsin,

fungal 1

Lipase Pancreatin 3

Cellulase Fungal 3

Wound

debridement

Streptokinase-

streptodornase

, trypsin,

bromelain

Bacterial,

animal, plant

1

Injection for

bruises,

inflammation

, etc.

Streptokinase,

trypsin

Bacterial,

animal

2

Paper test

strips for

diabetic

glucose

Glucose

oxidase,

peroxidase

Fungal, plant

2

Varied

clinical tests Numerous

Plat, animal,

microorganis

ms 3

Photograph

ic

Recovery of

silver from

spent film

Protease Bacterial

1

Textile

Desizing of

fabrics Amylase

Bacterial,

malt,

pancreatin 1

Protease

Bacterial,

fungal,

pancreatin 1

Vegetables

Liquefying

purees and

soups

Amylase Fungal

3

Dehydrated

vegetables,

restoring

flavor

Flavor Plants

4

Wine

Pressing,

clarification,

filtration

Pectinases Fungal

2

CHAPTER-1

14

Industry Application Enzyme Source* Extent

of Use+

High test

molasses Invertase Yeast

1

Miscellane

ous

Resolution

racemic

mixtures of

amino acids

Pectinases Fungal

4

Wall paper

removal Amylase Bacterial

3

Note: 1 - General and extensive industrial use

2 - Industrial use by some manufacturers

3 - Limited industrial use

4 - Laboratory or experimental use only

Later, the development of fermentation technology has specifically led to

the production of the high value enzymes by microbial fermentation with

selected strains and by using developed downstream processing facilities

and they were characterized and purified to a greater existent. In the

subsequently development use of recombinant gene technology has

improved manufacturing processes and enabled the commercialization of

novel enzymes that were not produced previously. In addition, the latest

developments in the modern biotechnology, introduced protein engineering

and directed evolution that have revolutionized the production of industrial

enzymes using the combination of various upstream and downstream

processes. So, these modern advances brought out the tailor-made enzymes

with added capabilities like quick adaption to new process conditions that

have generated the new applications for the greater use of enzymes in the

industries [42] as illustrated in Table-5 and the Figure-1. Hence we find

enzymes in this modern world as highly diversified product growing day

by day both in terms of size and complexity [21].

CHAPTER-1

15

Table-5. Expanded applications of enzymes in the modern industries [21]

Industry Enzyme class Applications

Detergent

(laundry and

dish wash)

Protease Protein stain removal

Amylase Starch stain removal

Lipase Lipid stain removal

Cellulase

Cleaing, color clarification, anti-

redeposting (cotton)

Mannanase

Mannanan stain removal

(reappearing stains

Starch and fuel

Amylase

Starch liquefaction and

saccharification

Amyglucosidase Saccharification

Pullulanase Saccharification

Glucose isomerase Glucose to fructose conversion

Cyclodextrin-

glycosyltransferase Cyclodextrin production

Xylanase

Viscosity reduction (fuel and

starch)

Protease protease (yeast nutrition fuel)

Food (including

dairy)

Protease

Milk clotting, infant formulas

(low allergenic), flavor

Lipase Cheese flavor

Pectin methyl

esterase Firming fruit based products

Pectinase Fruit based product

Transglutaminase Modify visco- elastic properties

Baking

Amylase

Bread softness and volume, flour

adjustment

Xylanase Dough conditioning

Lipase

Dough stability and conditioning

(in situ emulsifier)

Phospholipase

Dough stability and conditioning

(in situ emulsifier)

Glucose oxidase Dough strengthening

Lipoxygenase

Dough strengthening, bread

whitening

Protease Biscuits, cookies

Transglutaminase

Phytate digestibility- phosphorus

release

Animal feed Xylanase Digestibility

CHAPTER-1

16

Industry Enzyme class Applications

β-Glucanase Digestibility

Pectinase De-pectinization, mashing

Beverage

Amylase Juice treatment, low calorie beer

β-Glucanase Mashing

Acetolactate

decarboxylase Maturation (beer)

Laccase

Clarification (juice), flavor (beer),

cork stopper treatment

Textile

Cellulase Denim finishing, cotton softening

Amylase De-sizing

Pectate lyase Scouring

Catalase Bleach termination

Laccase Bleaching

Peroxidase Excess dye removal

Pulp and paper

Lipase Pitch control, contaminant control

Protease Biofilm removal

Amylase

Starch coating, de-inking,

drainage improvement

Xylanase Bleach termination

Cellulase

De-inking, drainage

improvement, fiber modification

Fats and oils

Lipase Transesterification

Phospholipase

De-gumming, lyso-lecithin

production

Organic

synthesis

Lipase

Resolution of chiral alcohols and

amides

Acylase

Synthesis of semisynthetic

pencillin

Nitrilase

Synthesis of enantiopure

carboxylic acids

Leather

Protease Unhearing, bating

Lipase De-picking

Personal care

Amyloglucosidase

Antimicrobial (combined with

glucose oxidase)

Glucose oxidase Bleaching, antimicrobial

Peroxidase Antimicrobial

CHAPTER-1

17

Figure 1. Enzymes used in the wet processing of fabric in Denim textile

industry

1.8 Objectives

Out of the many known industrial enzymes, discussed above, lipase

enzymes had been one of the versatile groups of biocatalysts used to carry

out various hydrolytic reactions in both aqueous and non-aqueous media

including their interface in the industries of dairy, textile, leather, pulp and

paper, fruit juices, fats and oils, health care [4] etc. It is also known that

lipases are isolated from various sources with wide range of substrate

specificity and non-specificity to catalyze the reaction involving chemo,

region and enantio-selective transformations. In a nutshell it is found that

lipases are used in the processing of fats and oils, detergents and degreasing

formulations, food processing, the synthesis of fine chemicals and

pharmaceuticals, paper manufacture and production of cosmetics and

pharmaceuticals, bakery, brewing, biofuels, leather, and general cleaning

[43-54] agents.

So the demand for lipases had been immense in the industries and at the

same time there had been various limitations posed by the special interfacial

specific catalytic activity, the enzyme physiochemical characters and the

diversity of lipases found in the nature. This has opened up the new areas

for research and the development towards the production and isolation of

novel lipases and their sources. With the existing information and the

CHAPTER-1

18

available resources, the objective for the research work were coined in the

following terms as

1. Collection of soil samples from the selected locations of Guntur,

Nagpur and Udaipur

2. Processing and screening of collected soil samples for isolation of

true lipase producing bacterial colonies

3. Optimization of culture media and physiological parameters for the

screening and isolation of best lipase producing bacterial isolates

4. Enhancement in lipase activity of the best bacterial soil isolates

using mutative strain improvement techniques

5. Identification and Characterization of the best lipase producing

bacterial soil isolates.

6. Optimal lipase production studies with the best lipase producing

bacterial soil isolates at their optimized process parameters using

lab scale batch fermenter