Enzymes microbial methodologies

The discovery and exploitation of enzymes in aldoxime-nitrile pathway nitrile hydratase, amidase, nitrilase, aldoxime dehydratase, etc., are shown along with the use of methodologies, such as organic chemistry, microbial screening by enrichment and acclimation culture techniques, enzyme purification, gene cloning, molecular screening by polymerase chain reaction (PCR). 

A notorious underestimation of the dynamic properties of microbial and cellular populations results from matching the duration of the respective batch cultivations to the relevant time constant of the biosystem under investigation. However, metabolic regulation of enzyme activities and fluxes often takes place on a time scale of seconds rather than days although the latter may also be true. It is therefore in the scope of promoting biotechnological research to adopt and develop appropriate experimental concepts, methodologies and equipment [397]. 

The microbial lipoxygenase was immobilised on polymer membrane, which was done according to the procedure in the methodological part. Examination of the obtained conjugate for protein contents was performed spectrophotometricaly in the way it was done for the enzyme immobilised on polymer granules. The relative activity of the immobilised enzyme is 65% and the protein contents 5.4 mg protein/g dry carrier. 

Combinatorial chemistry has become a powerful methodology for the construction of new compounds. Natural products, however, also have enormous potential as a source of new compounds. In particular, numerous useful microbial products have been isolated as antibiotics, herbicides, fungicides and enzyme-inhibitors. Moreover, microorganisms have provided various compounds with diverse bioactivities, such as immunomodulatory, antitumor and antihelmintical activities [1]. 

However, the above does not answer the main question how can one employ isolated enzymes for the preparation of surfactants In fact, the answer is simple Use hydrolytic enzymes in nonaqueous media. Indeed, many hydrolytic enzymes, such as lipases, proteases, and glycosidases, available in large quantities, are very robust and inexpensive, and do not require any cofactors to manifest their catalytic activity. As any other catalyst, enzymes cannot influence the equilibrium of a chemical reaction and therefore the removal of water from the reaction medium forces them to work in reverse, i.e., to synthesize a chemical bond rather than to break it. Consequently, there is a principal difference between microbial and enzymatic synthesis of surfactants regarding the type of enzymes involved and the reaction medium. The former is a biosynthetic process catalyzed by living microorganisms and as such dependent solely on their viability, whereas the latter is an organic synthesis whereby enzymes are used as substitutes for chemical catalysts. The two approaches are complementary not only in terms of the production methods but because the surfactant structures amenable to both methodologies are quite different. 

Sim, J.H., et al., 2007. Clostridium aceticum—a potential organism in catalyzing carbon monoxide to acetic acid application of response surface methodology. Enzyme and Microbial Technology 40 (5), 1234—1243. 

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