Production Of Enzyme Amylase By Bacillus Biology Essay

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Amylase is an enzyme that catalyses the breakdown of starch into sugars. Amylase is present in human saliva, where it begins the chemical process of digestion. Foods that contain much starch but little sugar, such as rice and potato, taste slightly sweet as they are chewed because amylase turns some of their starch into sugar in the mouth. Numerous microorganisms like Saccharomycopsis capsularia, Bacillus. coagulans, Bacillus sp. HOP-40, and Bacillus megatarium 16 M, have been used for a -amylase production by solid state fermentation (SSF) using agro-industrial residues. Amylases are important enzymes used in the food industry for hydrolysis of starch to yield glucose syrups amylase -rich flour and proper formation of dextrin during baking. Also they are employed in textile industry for removal of starch sizing. In the production of detergents, amylases are used to dissolve starches from fabrics. Amylases form 25% of the enzyme and are widely used in pharmaceuticals and sugar, brewing, textile, distilling and paper industries.

Amylases are important enzymes in nature and are synthesized microorganisms.

Two major classes of amylases have been identified in microorganisms, namely a-amylase and glucoamylase. a-Amylases are extracellular enzymes that randomly cleave the 1,4-a-D-glucosidic linkages between adjacent glucose units in the linear amylose chain. Glucoamylase hydrolyzes single glucose units from the nonreducing ends of amylose and amylopectin

They play a major role in clinical, medicinal and analytical chemistry and in baking, brewing, detergent, textile, paper and distilling industry. Large scale production of this enzyme is hence necessary at low costs. The cost of enzyme production in submerged fermentation (SmF) is high, which necessitates reduction in production cost by alternative methods. The contents of synthetic media are very expensive and these contents might be replaced with more economically available agricultural by-products for the reduction of cost of the medium. The use of agricultural wastes makes solid-state fermentation (SSF) an attractive alternative method have reported a-amylase production in solid-state fermentation with wheat bran and rice husk as substrates have checked the potential of coconut oil cake as substrate for the production of a-amylase using Aspergillus oryzae, a GRAS strain have described the selection of a suitable low cost fermentation medium for the production of a-amylase by using agricultural by-products. Glucoamylase production with an Aspergillus sp. has been reported using cheap rice flake manufacturing wastes as substrate.

The most effective amylases in industry are those that are thermostable. They are generally preferred as their application minimizes contamination risk and reduces reaction time, thus enabling considerable energy saving. Thermostable a-amylases are used for the liquefaction of starch at high temperature and thermolabile a-amylases are used for the saccharification of starch in baking. Babu and Satyanarayana have reported production of a-amylase by a thermophilic Bacillus sp. and optimization of culture conditions for maximum enzyme production. Suitability of thermophilic Bacillus coagulans for a-amylase production by solid-state fermentation in flasks, reactor and trays has been reported. In the present study a-amylase production from Bacillus cereus MTCC 1305 using solid-state fermentation has been investigated and the enzyme is reported to show activity at high temperature.

Bacillus species are known to produce large quantities of enzymes that can be purified for use in industry. Bacillus amyloliquefaciens is the source of alpha amylase used in starch hydrolysis, the protease subtilisin used with detergents, as well as a natural antibiotic protein barnase (a ribonuclease) and the BamH1 restriction enzyme used in DNA research.

There is immense use of amylases in the modern breadmaking industry. They are added as bread improver in form of malted barley to make the process faster Baker's yeast then feeds on the simple sugars in flour and converts it into the waste products of alcohol and CO2. Alpha amylases are important in brewing beer and liquor made from sugars derived from starch. In fermentation, yeast hydrolyzes sugars and excretes alcohol, which is not a waste in this case. In beer and some liquors, mashed grains or starch sources like potatoes are added the beginning of fermentation as a source of sugar.

For an enzyme to be used in the industry, it is important that the enzyme is able to withstand harsh conditions in the processes involved. Maintaining the temperature and pH which is bound to change during and industrial process may add to the costs of production and may make the process unfeasible. To address this problem, research is directed towards finding efficient and stable enzymes that can work on high temperatures and pH. In this lab, we are studying a-amylases that have an optimum at least of 70°C and pH 9.0.

Materials and methods : as per manual

Results and discussion:

The first step was to isolate the desired microorganism that produces α- amylase.

Among the seven selected Bacillus strains that showed the highest amylase synthesis, B. subtilis and B. cereus were selected because it gave a larger diameter zone of clearance and the highest relative amylolytic activity compared to the other species. These culture are further screened for amylase production by submerge fermentation using the hydrolysis method.

Results for screening of Bacillus species for amylase production

18th October 2012 Group 2 .

Bacillus Species

Group Number

Group1

Group2

Group3

Group4

Group5

Group6

Group7

Group8

B. sphaerius

-

-

-

-

-

-

-

N/A

B. pumilus

-

-

-

-

-

-

-

N/A

B. subtilis

++

++

++

++

+

N/A

++

++

B. megaterium

-

-

+

+

-

+

N/A

+

B.stearothermophilus

+

+

+

N/A

N/A

+

+

++

B. cereus

++

N/A

++

++

++

++

++

+++

B. mycoides

N/A

+

N/A

+

-

+

+

+

Legend: - No effect

+ Small effect

++ Medium effect

+++ Large effect

Conclusions:

Amylases have been obtained as a result of screening of Bacillus sp. strains, isolated from different natural substrates. Seven thermostable a-amylases demonstrated 100% of their activities after 60 min of incubation at 100 C. These enzymes showed maximal activities at pH 9.0 and exhibited 10-30% of activities at pH 11.0 and 12.0. These properties make them promising for future research in the pharmaceutical industry but also in other industries e.g. food, cloth for a sustainable development.

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