novel strategy for enzymatic synthesis of 4-hydroxyisoleucine: identification of an enzyme possessing HMKP (4-hydroxy-3-methyl-2-keto-pentanoate) aldolase activity

A two-step enzymatic synthesis process of 4-hydroxyisoleucine is suggested. In the first step, the aldol condensation of acetaldehyde and α-ketobutyrate catalyzed by specific aldolase results in the formation of 4-hydroxy-3-methyl-2-keto-pentanoate (HMKP). In the second step, amination of HMKP by th...

Full description

Saved in:
Bibliographic Details
Published in:FEMS microbiology letters 2007-08, Vol.273 (1), p.70-77
Main Authors: Smirnov, Sergey V, Samsonova, Natalya N, Novikova, Anna E, Matrosov, Nikolay G, Rushkevich, Natalya Y, Kodera, Tomohiro, Ogawa, Jun, Yamanaka, Hiroyuki, Shimizu, Sakayu
Format: Article
Language:English
Subjects:
4-hydroxyisoleucine
Acetaldehyde
Acetaldehyde - metabolism
Aldehydes
Aldolase
Amination
Amino acids
Arthrobacter
Arthrobacter - enzymology
Arthrobacter simplex
Bacterial Proteins - genetics
Bacterial Proteins - isolation & purification
Bacterial Proteins - metabolism
Base Sequence
biotransformation
Butyrates - metabolism
Chain branching
Chains
DNA, Bacterial - chemistry
DNA, Bacterial - genetics
Enzymatic synthesis
Enzymes
Fructose-Bisphosphate Aldolase - genetics
Fructose-Bisphosphate Aldolase - isolation & purification
Fructose-Bisphosphate Aldolase - metabolism
Gene sequencing
Glutamic acid
Glutamic Acid - metabolism
HpcH/HpaI aldolase family
Isoleucine - analogs & derivatives
Isoleucine - metabolism
Microbiology
Molecular Sequence Data
noninsulin-dependent diabetes mellitus
Sequence Analysis, DNA
Synthesis
Transaminases - metabolism
type II diabetes
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:A two-step enzymatic synthesis process of 4-hydroxyisoleucine is suggested. In the first step, the aldol condensation of acetaldehyde and α-ketobutyrate catalyzed by specific aldolase results in the formation of 4-hydroxy-3-methyl-2-keto-pentanoate (HMKP). In the second step, amination of HMKP by the branched-chain amino acid aminotransferase leads to synthesis of 4-hydroxyisoleucine. An enzyme possessing HMKP aldolase activity (asHPAL) was purified 2500-fold from a crude extract of Arthrobacter simplex strain AKU 626. Sequencing of the asHPAL structural gene showed that the purified enzyme belongs to the HpcH/HpaI aldolase family. The 4-hydroxyisoleucine was synthesized in vitro from acetaldehyde, α-ketobutyrate and l-glutamate using a coupled aldolase/branched-chain amino acid aminotransferase bienzymatic reaction.
ISSN:0378-1097
1574-6968
DOI:10.1111/j.1574-6968.2007.00783.x