Substrate inhibition mode of ω-transaminase from Vibrio fluvialis JS17 is dependent on the chirality of substrate

Jong Shik Shin, Byung Gee Kim

Research output: Contribution to journalArticle

44 Citations (Scopus)

Abstract

Substrate inhibition is a common phenomenon in enzyme chemistry, which is observed only with a fast-reacting substrate enantiomer. We report here for the first time substrate inhibition of an enantioselective enzyme by both substrate enantiomers. The enantioselective substrate inhibition, i.e., different mode of inhibition by each substrate enantiomer, of (S)-specific ω-transaminase was found with various chiral amines. A kinetic model based on ping-pong bi-bi mechanism has been developed and kinetic parameters were measured. The kinetic model reveals that the inhibition by (R)-amine results from formation of Michaelis complex with enzyme-pyridoxal 5′-phosphate, whereas the inhibition by (S)-amine results from the formation of the complex with enzyme-pyridoxamine 5′-phosphate. Substrate inhibition constants (KSI) of each (S)-enantiomer of four chiral amines showed a linear correlation with those of cognate (R)-amines. Such a correlation was also found between the KSI values and Michaelis constants of (S)-amines. These correlations indicate that recognition mechanisms and active site structures of both enzyme-pyridoxal 5′-phosphate, enzyme-pyridoxamine 5′-phosphate are similar. Taken together with the results, high propensity for non-productive substrate binding strongly suggests that binding pockets of the ω-transaminase is loosely defined, which accounts for the enantioselective substrate inhibition.

Original languageEnglish
Pages (from-to)832-837
Number of pages6
JournalBiotechnology and Bioengineering
Volume77
Issue number7
DOIs
Publication statusPublished - 2002 Mar 30

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Vibrio
Chirality
Transaminases
Amines
Enzyme inhibition
Substrates
Enzymes
Enantiomers
Pyridoxal Phosphate
Phosphates
Pyridoxamine
Catalytic Domain
Kinetics
Kinetic parameters

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology

Cite this

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title = "Substrate inhibition mode of ω-transaminase from Vibrio fluvialis JS17 is dependent on the chirality of substrate",
abstract = "Substrate inhibition is a common phenomenon in enzyme chemistry, which is observed only with a fast-reacting substrate enantiomer. We report here for the first time substrate inhibition of an enantioselective enzyme by both substrate enantiomers. The enantioselective substrate inhibition, i.e., different mode of inhibition by each substrate enantiomer, of (S)-specific ω-transaminase was found with various chiral amines. A kinetic model based on ping-pong bi-bi mechanism has been developed and kinetic parameters were measured. The kinetic model reveals that the inhibition by (R)-amine results from formation of Michaelis complex with enzyme-pyridoxal 5′-phosphate, whereas the inhibition by (S)-amine results from the formation of the complex with enzyme-pyridoxamine 5′-phosphate. Substrate inhibition constants (KSI) of each (S)-enantiomer of four chiral amines showed a linear correlation with those of cognate (R)-amines. Such a correlation was also found between the KSI values and Michaelis constants of (S)-amines. These correlations indicate that recognition mechanisms and active site structures of both enzyme-pyridoxal 5′-phosphate, enzyme-pyridoxamine 5′-phosphate are similar. Taken together with the results, high propensity for non-productive substrate binding strongly suggests that binding pockets of the ω-transaminase is loosely defined, which accounts for the enantioselective substrate inhibition.",
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Substrate inhibition mode of ω-transaminase from Vibrio fluvialis JS17 is dependent on the chirality of substrate. / Shin, Jong Shik; Kim, Byung Gee.

In: Biotechnology and Bioengineering, Vol. 77, No. 7, 30.03.2002, p. 832-837.

Research output: Contribution to journalArticle

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