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

Ritu Sareen, Uwe T. Bornscheuer, Prashant Mishra

Research output: Contribution to journalArticle

21 Citations (Scopus)

Abstract

The synthesis of the analgesic dipeptide kyotorphin precursor (Bz-Tyr-Arg-NH 2) 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). α-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-NH 2) at a concentration 400 mM 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.

Original languageEnglish
Pages (from-to)1-5
Number of pages5
JournalJournal of Molecular Catalysis B: Enzymatic
Volume32
Issue number1-2
DOIs
Publication statusPublished - 22-12-2004

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kyotorphin
Nucleophiles
Bacilli
Organic solvents
Chymotrypsin
Peptide Hydrolases
Acetonitrile
Dipeptides
Amides
Analgesics
Esters
Hydrolysis
Bacillus licheniformis

All Science Journal Classification (ASJC) codes

  • Catalysis
  • Bioengineering
  • Biochemistry
  • Process Chemistry and Technology

Cite this

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abstract = "The synthesis of the analgesic dipeptide kyotorphin precursor (Bz-Tyr-Arg-NH 2) 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). α-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-NH 2) at a concentration 400 mM 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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Synthesis of kyotorphin precursor by an organic solvent-stable protease from Bacillus licheniformis RSP-09-37. / Sareen, Ritu; Bornscheuer, Uwe T.; Mishra, Prashant.

In: Journal of Molecular Catalysis B: Enzymatic, Vol. 32, No. 1-2, 22.12.2004, p. 1-5.

Research output: Contribution to journalArticle

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