Abstract
Microorganisms that are capable of (S)-enantioselective transamination of chiral amines were isolated from soil samples by selective enrichment using (S)-α-methylbenzylamine ((S)-α-MBA) as a sole nitrogen source. Among them, Klebsiella pneumoniae JS2F, Bacillus thuringiensis JS64, and Vibrio fluvialis JS17 showed good ω-transaminase (ω-TA) activities and the properties of the ω-TAs were investigated. The induction level of the enzyme was strongly dependent on the nitrogen source for the strains, except for V. fluvialis JS17. All the ω-TAs showed high enantioselectivity (E>50) toward (S)-α-MBA and broad amino donor specificities for arylic and aliphatic chiral amines. Besides pyruvate, aldehydes such as propionaldehyde and butyraldehyde showed good amino acceptor reactivities. All the ω-TAs showed substrate inhibition by (S)-α-MBA above 200 mM. Moreover, substrate inhibition by pyruvate above 10 mM was observed for ω-TA from V. fluvialis JS17. In the case of product inhibition, acetophenone showed much greater inhibitions than L-alanine for all ω-TAs. Comparison of the enzyme properties indicates that ω-transaminase from V. fluvialis JS17 is the best one for both kinetic resolution and asymmetric synthesis to produce enantiomerically pure chiral amines. Kinetic resolution of sec-butylamine (20 mM) was done under reduced pressure (150 Torr) to selectively remove an inhibitory product (2-butanone) using the enzyme from V. fluvialis JS17. Enantiomeric excess of (R)-sec-butylamine reached 94.7% after 12 h of reaction.
| Original language | English |
|---|---|
| Pages (from-to) | 1782-1788 |
| Number of pages | 7 |
| Journal | Bioscience, Biotechnology and Biochemistry |
| Volume | 65 |
| Issue number | 8 |
| DOIs | |
| Publication status | Published - 2001 Aug 1 |
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All Science Journal Classification (ASJC) codes
- Biotechnology
- Analytical Chemistry
- Biochemistry
- Applied Microbiology and Biotechnology
- Molecular Biology
- Organic Chemistry
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Comparison of the ω-transaminases from different microorganisms and application to production of chiral amines. / Shin, Jong Shik; Kim, Byung Gee.
In: Bioscience, Biotechnology and Biochemistry, Vol. 65, No. 8, 01.08.2001, p. 1782-1788.Research output: Contribution to journal › Article
TY - JOUR
T1 - Comparison of the ω-transaminases from different microorganisms and application to production of chiral amines
AU - Shin, Jong Shik
AU - Kim, Byung Gee
PY - 2001/8/1
Y1 - 2001/8/1
N2 - Microorganisms that are capable of (S)-enantioselective transamination of chiral amines were isolated from soil samples by selective enrichment using (S)-α-methylbenzylamine ((S)-α-MBA) as a sole nitrogen source. Among them, Klebsiella pneumoniae JS2F, Bacillus thuringiensis JS64, and Vibrio fluvialis JS17 showed good ω-transaminase (ω-TA) activities and the properties of the ω-TAs were investigated. The induction level of the enzyme was strongly dependent on the nitrogen source for the strains, except for V. fluvialis JS17. All the ω-TAs showed high enantioselectivity (E>50) toward (S)-α-MBA and broad amino donor specificities for arylic and aliphatic chiral amines. Besides pyruvate, aldehydes such as propionaldehyde and butyraldehyde showed good amino acceptor reactivities. All the ω-TAs showed substrate inhibition by (S)-α-MBA above 200 mM. Moreover, substrate inhibition by pyruvate above 10 mM was observed for ω-TA from V. fluvialis JS17. In the case of product inhibition, acetophenone showed much greater inhibitions than L-alanine for all ω-TAs. Comparison of the enzyme properties indicates that ω-transaminase from V. fluvialis JS17 is the best one for both kinetic resolution and asymmetric synthesis to produce enantiomerically pure chiral amines. Kinetic resolution of sec-butylamine (20 mM) was done under reduced pressure (150 Torr) to selectively remove an inhibitory product (2-butanone) using the enzyme from V. fluvialis JS17. Enantiomeric excess of (R)-sec-butylamine reached 94.7% after 12 h of reaction.
AB - Microorganisms that are capable of (S)-enantioselective transamination of chiral amines were isolated from soil samples by selective enrichment using (S)-α-methylbenzylamine ((S)-α-MBA) as a sole nitrogen source. Among them, Klebsiella pneumoniae JS2F, Bacillus thuringiensis JS64, and Vibrio fluvialis JS17 showed good ω-transaminase (ω-TA) activities and the properties of the ω-TAs were investigated. The induction level of the enzyme was strongly dependent on the nitrogen source for the strains, except for V. fluvialis JS17. All the ω-TAs showed high enantioselectivity (E>50) toward (S)-α-MBA and broad amino donor specificities for arylic and aliphatic chiral amines. Besides pyruvate, aldehydes such as propionaldehyde and butyraldehyde showed good amino acceptor reactivities. All the ω-TAs showed substrate inhibition by (S)-α-MBA above 200 mM. Moreover, substrate inhibition by pyruvate above 10 mM was observed for ω-TA from V. fluvialis JS17. In the case of product inhibition, acetophenone showed much greater inhibitions than L-alanine for all ω-TAs. Comparison of the enzyme properties indicates that ω-transaminase from V. fluvialis JS17 is the best one for both kinetic resolution and asymmetric synthesis to produce enantiomerically pure chiral amines. Kinetic resolution of sec-butylamine (20 mM) was done under reduced pressure (150 Torr) to selectively remove an inhibitory product (2-butanone) using the enzyme from V. fluvialis JS17. Enantiomeric excess of (R)-sec-butylamine reached 94.7% after 12 h of reaction.
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U2 - 10.1271/bbb.65.1782
DO - 10.1271/bbb.65.1782
M3 - Article
C2 - 11577718
AN - SCOPUS:0035433593
VL - 65
SP - 1782
EP - 1788
JO - Bioscience, Biotechnology and Biochemistry
JF - Bioscience, Biotechnology and Biochemistry
SN - 0916-8451
IS - 8
ER -