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Microbial production of Carboxypeptidase G2.

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Relation http://ir.cftri.com/11450/
 
Title Microbial production of Carboxypeptidase G2.
 
Creator Sachin, K.
 
Subject 04 Microbiology
05 Enzymes
 
Description Enzymes that cleave carboxy terminal of a protein are known as Carboxypeptidases (EC number 3.4.16 - 3.4.18). There are many types of carboxypeptidases which are classified based on which amino acid it act upon. Carboxypeptidase G2 (EC 3.4.17.21) is a metallo-carboxypeptidase that is predominantly expressed as a membrane-bound enzyme, it also exists in a soluble form. It Hydrolyses a peptide bonds in Ac-Asp-Glu, Asp-Glu, and Glu-Glu, but also γ-glutamyl bonds in γ-Glu-Glu, and folylpoly-γ-glutamate at c-terminal of the peptide bond
Microbial enzymes are produced on a large scale is been carried out by fermentation techniques for many decades. Fermentation could be of two type’s solid state or submerged stated fermentation. When mold are used it could be either solid state fermentation, when bacteria or yeast are used it is submerged state fermentation. The growth conditions for the organism have to be optimized for higher yield of enzymes. This could be done by using statistical method. Therapeutic enzymes have to be free from all the impurities before it could be used. Chromatography techniques could be used to obtain high degree of purification.
Application:
Folic acid derivate (folate) is important in biochemical synthesis of DNA. Depleting folate reserves will lead to inhibition of DNA synthesis and thereby can inhibit cell division. Folate depletion can be achieved directly by enzymatic cleavage of glutamate moiety present in the molecule (Roger et al., 1985). The groups of enzymes capable of such hydrolysis have been designated as Carboxypeptidase G and various forms of these enzymes have been isolated from number of bacteria and molds.
Carboxypeptidase G2 also acts upon methotrexate, a prodrug, which is an analog of folate molecule. Prodrug is inactive compound in a wide range, which upon modification is
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converted in to active drug. The drug is active by the action of an enzyme present in the host cell or can be activated by microbial enzymes. Prodrug/enzyme complex mediate toxicity by disruption of DNA replication, which occurs at differentially high rates in tumor cells compared to most normal cells.
 
Contributor Manonmani, H. K.
 
Date 2012
 
Type Student Project Report
NonPeerReviewed
 
Format application/pdf
 
Language en
 
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Identifier http://ir.cftri.com/11450/1/sachink.pdf
Sachin, K. (2012) Microbial production of Carboxypeptidase G2. [Student Project Report] (Submitted)