This articles is based on information regarding how to produce microbial enzymes, methods of enzyme purification including sources and application of microbial enzymes.

1. INTERNATIONALISLAMIC
UNIVERSITY,ISLAMABAD
ENZYMOLOGY ASSIGNMENT
NKJ
3-20-2015
SUBMITTED TO: DR ZEHRA
SUBMITTED BY: NIDA JAVED
REG # 516-FBAS-BSBT (6A)-F13

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ENZYMES AND BIOTECHNOLOGY
MICROBIAL ENZYMES:
Possible industrial application of enzymes to carry out certain reactions apart from the cell dates
back many centuries and was practically used long before the activity of enzymes was known.
Examples of ancient use of enzymes are usage of barley malt for starch conversion in brewing,
and of droppings for bating of hides in leather making. Coarse preparations from some animal
tissues such as pancreas and stomach mucosa, or from plant tissues for example, malt and papaya
fruit, were prepared which found industrial applications in the textile, leather, brewing, and other
industries. Once the positive results of applying such enzyme preparations were entrenched, a
search opened for better, inexpensive, and more easily accessible sources of such enzymes. It was
observed that some microorganisms produce enzymes identical in action to the amylases of malt
and pancreas, or to the proteases of the pancreas and papaya fruit. This led to the advancement of
processes for producing such microbial enzymes on industrial scale.
INDUSTRIAL USE OF MICROBIAL ENZYMES
Dr. Jokichi Takamine (1894, 1914) was the first person to recognize the technological prospects
of cultivated enzymes and to introduce them to industry. He was concerned with fungal enzymes,
whereas Boidin and Effront (1917) in France discovered the production of bacterial enzymes about
20 years later. Industrial advancement in this field during the last decades has been so vast that,
for many purposes, microbial cultivated enzymes have replaced the animal or plant enzymes. For
example, in textile designing, bacterial amylase has broadly replaced malt or pancreatin. Currently,
relatively limited number of microbial enzymes have found commercial application, but the
number is rising, and the field will assuredly be much broaden in the future.
PRODUCTION OF MICROBIAL ENZYMES
1) FUNGAL ENZYMES:
For fungal enzymes, alterations of Dr. Takamine's mold bran process have consistently been
employed. In this process, the mold is cultivated on the outer surface of a solid substrate. Takamine
used wheat bran and this has been observed as the most suitable basic substrate although other
fibrous materials can be employed. Other ingredients may be added, such as nutrient salts, acid or

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NIDA JAVEDREG # 516-BSBT (6A)-F13
buffer to regulate the pH, soy bean meal or beet cosettes to stimulate enzyme production. In one
alteration of the bran procedure, the bran is steamed for sterilization, cooled, inoculated with the
mold spores, and spread out on trays (Underkofler et al., 1947; Forbath, 1957). Incubation takes
place in chambers where the temperature and humidity are restrained by circulated air. Instead of
trays for incubation, Takamin and other producers, at one time used slowly rotating drums. Usually
tray incubation gives more rapid growth and enzyme production.
TABLE 1
Some Commercial Enzyme and Source Microorganisms
SOURCE ENZYME MICROORGANISM
Fungal
Amylases
Glucosidases
Proteases
Pectinases
Glucose oxidase
Catalase
Aspergillus oryzae
Aspergillus flavus
Aspergillus niger
Aspergillus niger
Penicillium notatum
Aspergillus niger
Bacterial
Amylases
Proteases
Penicillinase
Bacillus subtilis
Yeast Invertase Lactase Saccharomyces cerevisiae
Saccharomyces fragilis
Resource: http://aem.asm.org/content/6/3/212.full.pdf
2) BACTERIAL ENZYMES:
Bacterial enzymes have been and are also produced by the bran process. Latterly the process
originally invented by Boidin and Effront (1917) was most broadly employed (Wallerstein, 1939).
In this process, the bacteria are cultivated in specific culture vessels as a pellicle on the surface of
thin layers of liquid medium, the content of which is adjusted for maximum production of the
required enzyme. Different strains of Bacillus subtilis and different media are employed,
depending on whether bacterial amylase or protease is required.

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METHODS OF ENZYME PURIFICATION:
animal, plant,
microbe
chemical method:
Detergent, alkali-
Enzyme method:
lysozyme
centrifugation,
filteration
Heat treatment,
Salts
Ion exchange
chromatography,
Gel
chromatography
therapeutic
enzyme, Industrial
enzyme
CELL SOURCE
CELL DISRUPTION
DEBRIS REMOVAL
INITIAL
PURIFICATION
HIGH RESOLUTION
PURIFICATION
PURIFIED ENZYME

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MICROBIAL ENZYMES
TABLE 2
Uses of Industrial Enzyme Preparations
INDUSTRY APPLICATION ENZYME SOURCE EXTENT
OF USE
Baking and milling Bread baking Amylase
Protease
fungal, malt
fungal
2
1
Beer Mashing
Chillproofing
Oxygen removal
Amylase
Protease
Glucose oxidase
malt, bacterial
papain, bromelain,
pepsin, fungal,
bacterial
fungal
1
1
3
Carbonated beverages Oxygen removal Glucose oxidase fungal 3
Cereals Precooked baby foods
Breakfast foods
condiments
Amylase
Amylase
Protease
Malt, fungal
Malt, fungal
Papain, bromelain,
pepsin, fungal,
bacterial
2
2
2
Chocolate, cocoa
Coffee
Syrups
Coffee bean
fermentation
Coffee concenterates
amylase
Pectinase
Pectinase,
hemicellulase
Fungal, bacterial
Fungal
Fungal
2
2
2
Confectionery, candy
Soft center candies and
fondants
Sugar recovery from
scrap candy
Invertase
Amylase
Yeast
Bacterial, fungal
2
3
Dairy Cheese production Rennin Animal 1
3
2
2
4
Milk, sterilization with
peroxide
Catalase Liver,bacterial
Milk, prevention of
oxidation flavor
Protease Pancreatin
Milk, protein
hydrolyzates
Protease Papain,bromelain,
pancreatin,fungal,
bacterial
Evaporatedmilk,
stabilization
Protease Pancreatin,pepsin,
fungal,bromelain
Fungal
Distilled beverages Mashing Amylase
Malt, fungal,
bacterial
1

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Dry cleaning, laundry Spot removal Protease,lipase,
amylase
Bacterial, pancreatin,
fungal
1
Pharmaceuticaland clinical Digestive aids Amylase
Protease
Lipase
Cellulase
Fungal,pancreatin
Papain,pancreatin,
bromelain,pepsin,
fungal
Pancreatin
Fungal
1
1
3
3
1Wound debridement Streptokinase-
streptodornase,
trypsin, bromelain
Bacterial, animal,
plant
Injection for bruises,
inflammation
Streptokinase, trypsin Bacterial, animal 2
http://aem.asm.org/content/6/3/212.full.pdf
* Where one of optional sourcespredominatesithasbeenitalicized.
t 1 General andextensiveindustrial use.
2 Industrial use bysome manufacturers.
3 Limitedindustrial use.
4 Laboratoryor experimental use only.

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REFERENCES:
 http://aem.asm.org/content/6/3/212.full.pdf
 http://www.biochemistry.org/portals/0/education/docs/basc03_full.pdf