Can I read Construction and co-expression of a polycistronic plasmid encoding carbonyl reductase and glucose dehydrogenase for production of ethyl (S)-4-chloro-3-hydroxybutanoate on EtoBox?

Construction and co-expression of a polycistronic plasmid encoding carbonyl reductase and glucose dehydrogenase for production of ethyl (S)-4-chloro-3-hydroxybutanoate by Qi Ye; Hou Cao; Ming Yan; Fei Cao; Yueyuan Zhang; Ximu Li; Lin Xu; Yong Chen; Jian Xiong; Pingkai Ouyang; Hanjie Ying is a scholarly article available to read on EtoBox.

What is Construction and co-expression of a polycistronic plasmid encoding carbonyl reductase and glucose dehydrogenase for production of ethyl (S)-4-chloro-3-hydroxybutanoate about?

Biocatalysis of ethyl 4-chloro-3-oxobutanoate (COBE) to ethyl (S)-4-chloro-3-hydroxybutanoate [(S)-CHBE] was carried out using Escherichia coli co-expressing a carbonyl reductase gene from Pichia stipitis and a glucose dehydrogenase gene from Bacillus megaterium. An efficient polycistronic plasmid with a high-level of enzyme co-expression was constructed by changing the order of the genes, altering the Shine-Dalgarno (SD) regions, and aligned spacing (AS) between the SD sequence and the translation initiation codon. The optimal SD sequence was 5-TAAGGAGG-3, and the optimal AS distance was eight nucleotides. Asymmetric reduction of COBE to (S)-CHBE with more than 99% enantiomeric excess was demonstrated by transformants, using a water/ethyl caprylate system. The recombinant cells produced 1260 mM product in the organic phase, and the total turnover number, defined as moles (S)-CHBE formed per mole NADP(+), was 12,600, which was more than 10-fold higher than in aqueous systems.

Author

Qi Ye; Hou Cao; Ming Yan; Fei Cao; Yueyuan Zhang; Ximu Li; Lin Xu; Yong Chen; Jian Xiong; Pingkai Ouyang; Hanjie Ying

Publisher

Elsevier Science; Elsevier ; Elsevier BV (ISSN 0960-8524)

Published

2010

Language

EN