Designing of a Cofactor Self-Sufficient Whole-Cell Biocatalyst System for Production of 1,2-Amino Alcohols from Epoxides

Abstract

Optically pure 1,2-amino alcohols are highly valuable products as intermediates for chiral pharmaceutical products. Here we designed an environmentally friendly non-natural biocatalytic cascade for efficient synthesis of 1,2-amino alcohols from cheaper epoxides. A redesignated ω-transaminase PAKω-TA was tested and showed good bioactivity at a lower pH than other reported transaminases. The cascade was efficiently constructed as a single one-pot E. coli recombinant, by coupling SpEH (epoxide hydrolase), MnADH (alcohol dehydrogenase), and PAKω-TA. Furthermore, RBS regulation strategy was used to overcome the rate limiting step by increasing expression of MnADH. For cofactor regeneration and amino donor source, an interesting point was involved as that a cofactor self-sufficient system was designed by expression of GluDH. It established a “bridge” between the cofactor and the cosubstrate, such that the cofactor self-sufficient system could release cofactor (NADP+) and cosubstrate (l-Glutamine) regenerated simultaneously. The recombinant E. coli BL21 (SGMP) with cofactor self-sufficient whole-cell cascade biocatalysis showed high ee value (>99%) and high yield, with 99.6% conversion of epoxide (S)-1a to 1,2-amino alcohol (S)-1d in 10 h. It further converted (S)-2a–5a to (S)-2d–5d with varying conversion rates ranging between 65–96.4%. This study first provides one-step synthesis of optically pure 1,2-amino alcohols from (S)-epoxides employing a synthetic redox-self-sufficient cascade.

Efficient Synthesis of (R)-(+)-Perillyl Alcohol From (R)-(+)-Limonene Using Engineered Escherichia coli Whole Cell Biocatalyst

“…NADH addition can obviously increase ( R )-(+)-perillyl alcohol yield ( Figure 4C ). Since NADH is costly, we integrated a NADH generating pathway, such as, formate dehydrogenase (FDH) for the regeneration of NADH ( Liu et al, 2019 ; Chen et al, 2021 ). FDH from Candida boidinii was expressed and strain 10 was constructed.…”

Improved Bio-Synthesis of 2,5-bis(hydroxymethyl)furan by Burkholderia contaminans NJPI-15 With Co-substrate

“…Glutamine plays a pleiotropic role in cell function, contributing to processes such as energy synthesis, macromolecular synthesis, mTOR activation, and reactive oxygen balance (Sayed et al, 2019). Liu et al (2019) reported that a cofactor self-sufficient system was designed for enhanced 1,2-amino alcohols production in E. coli BL21 by overexpressing a glutamate dehydrogenase. Efficient biosynthesis of (S)-1,2-amino alcohols with respective high yields without addition of external cofactors and amino donors.…”

Biochemical Characterization and Structural Insight into Interaction and Conformation Mechanisms of Serratia marcescens Lysine Decarboxylase (SmcadA)

“…The recombinant strains were constructed using a modified method by Liu et al [38]. Briefly, the Serratia marcescens W2.3 genome sequence was used to design primers.…”