In Vitro and in Vivo One-Pot Deracemization of Chiral Amines by Reaction Pathway Control of Enantiocomplementary ω-Transaminases

Sang Woo Han, Youngho Jang, Jong Shik Shin

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Biocatalytic cascade conversion of racemic amines into optically pure ones using enantiocomplementary ω-transaminases (ω-TAs) has been developed by thermodynamic and kinetic control of reaction pathways where 12 competing reactions occur with pyruvate and isopropylamine used as cosubstrates. Thermodynamic control was achieved under reduced pressure for selective removal of a coproduct (i.e., acetone), leading to elimination of six undesirable reactions. Engineered orthogonality in substrate specificities of ω-TAs was exploited for kinetic control, enabling suppression of four additional reactions. Taken together, the net reaction pathway could be directed to two desired reactions (i.e., oxidative deamination of R-amine and reductive amination of the resulting ketone into antipode S-amine). This strategy afforded one-pot deracemization of various chiral amines with >99% eeS and 85-99% reaction yields of the resulting S-amine products. The in vitro cascade reaction could be successfully implemented in a live microbe using glucose or l-threonine as a cheap amino acceptor precursor, demonstrating a synthetic metabolic pathway enabling deracemization of chiral amines which has never been observed in living organisms.

Original languageEnglish
Pages (from-to)6945-6954
Number of pages10
JournalACS Catalysis
Volume9
Issue number8
DOIs
Publication statusPublished - 2019 Aug 2

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Transaminases
Amines
Thermodynamics
Amination
Kinetics
Threonine
Acetone
Ketones
Pyruvic Acid
Glucose
Substrates

All Science Journal Classification (ASJC) codes

  • Catalysis
  • Chemistry(all)

Cite this

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title = "In Vitro and in Vivo One-Pot Deracemization of Chiral Amines by Reaction Pathway Control of Enantiocomplementary ω-Transaminases",
abstract = "Biocatalytic cascade conversion of racemic amines into optically pure ones using enantiocomplementary ω-transaminases (ω-TAs) has been developed by thermodynamic and kinetic control of reaction pathways where 12 competing reactions occur with pyruvate and isopropylamine used as cosubstrates. Thermodynamic control was achieved under reduced pressure for selective removal of a coproduct (i.e., acetone), leading to elimination of six undesirable reactions. Engineered orthogonality in substrate specificities of ω-TAs was exploited for kinetic control, enabling suppression of four additional reactions. Taken together, the net reaction pathway could be directed to two desired reactions (i.e., oxidative deamination of R-amine and reductive amination of the resulting ketone into antipode S-amine). This strategy afforded one-pot deracemization of various chiral amines with >99{\%} eeS and 85-99{\%} reaction yields of the resulting S-amine products. The in vitro cascade reaction could be successfully implemented in a live microbe using glucose or l-threonine as a cheap amino acceptor precursor, demonstrating a synthetic metabolic pathway enabling deracemization of chiral amines which has never been observed in living organisms.",
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In Vitro and in Vivo One-Pot Deracemization of Chiral Amines by Reaction Pathway Control of Enantiocomplementary ω-Transaminases. / Han, Sang Woo; Jang, Youngho; Shin, Jong Shik.

In: ACS Catalysis, Vol. 9, No. 8, 02.08.2019, p. 6945-6954.

Research output: Contribution to journalArticle

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