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Synthesis of kyotorphin precursor by an organic solvent-stable protease from bacillus licheniformis RSP-09-37

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Author: Sareena, Ritu; Bornscheuerb, Uwe T; Mishra, Prashant

Advisor: Advisor

Date: 2004

Publisher:
Citation: Journal of

Series/Report no.:
Item Type: Article

Keywords: Protease; Acetonitrile; Peptide synthesis; Kyotorphin; S/H ratio

Abstract: The synthesis of the analgesic dipeptide kyotorphin precursor (Bz-Tyr-Arg-NH2) was studied under kinetically controlled conditions in 10–90% (v/v) aqueous-acetonitrile media at −20 ◦C using a novel protease obtained from the cell free supernatant of a Bacillus licheniformis mutant strain (RSP-09-37).Chymotrypsin (CT) was used for comparison. The conditions for maximum yield of kyotorphin precursor synthesis were optimized using CT by varying the type of nucleophile (amide and ester), nucleophile concentration and nucleophile to acyl donor ratio. The nucleophile (Arg-NH2) at a concentration 400mM and nucleophile to acyl donor ratio 1:40 was found to be optimum for kyotorphin precursor synthesis. The protease from B. licheniformis RSP-09-37 was stable even at 90% acetonitrile concentration and allowed for a significantly higher synthesis over hydrolysis ratio (S/H ratio) of 15.6 compared to only 3.0 found for CT at −20 ◦C.
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Shankar B. Chavan
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Shankar B. Chavan
Computer Applications Division
Central Library, IIT Delhi
shankar.chavan@library.iitd.ac.in
NDLTD
Shodhganga
NDL
ePrints@IISc
etd@IISc
IR@IIT Bombay
NewsClips @IITD
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