Highly efficient asymmetric reduction of ketopantolactone to d-(−)-pantolactone by Escherichia coli cells expressing recombinant conjugated polyketone reductase and glucose dehydrogenase in a fed-batch biphasic reaction system

2020 ◽  
Vol 5 (3) ◽  
pp. 531-538
Author(s):  
Xiaolin Pei ◽  
Jiapao Wang ◽  
Haoteng Zheng ◽  
Pengfei Cheng ◽  
Yifeng Wu ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Asymmetric Reduction ◽  
Glucose Dehydrogenase ◽  
Reaction System ◽  
Fed Batch ◽  
Highly Efficient ◽  
Escherichia Coli Cells ◽  
System P ◽  
Biphasic Reaction

Enantiopure d-(−)-pantolactone was efficiently synthesized by Escherichia coli cells expressing recombinant CduCPR and BsuGDH in a fed-batch biphasic reaction system.

Asymmetric Synthesis of Ethyl (R)-4-Chloro-3-Hydroxybutanoate with a Two Recombinant Escherichia coli System

2013 ◽  
Vol 690-693 ◽  
pp. 1188-1192
Author(s):  
Ke Ju Jing ◽  
Wen Ting Ban
Keyword(s):  
Escherichia Coli ◽  
Butyl Acetate ◽  
Asymmetric Reduction ◽  
Glucose Dehydrogenase ◽  
Reaction System ◽  
Phase Reaction ◽  
Aldehyde Reductase ◽  
Two Phase ◽  
E Coli ◽  
Final Yield

The asymmetric reduction of ethyl 4-chloro-acetoacetate (CAAE) to ethyl (R)-4-chloro-3- hydroxybutanoate (R-CHBE) biocatalysed by the aldehyde reductase of Sporobolomyces salmonicolor expressed in E. coli M15 (pQE30-alr) in combination with regeneration of NADPH by the glucose dehydrogenase of Bacillus megaterium expressed in E. coli M15 (pQE30-gdh) was reported. The bioreduction was carried out in a two-phase reaction system with n-butyl acetate as an organic solvent. Bioconversion of 300 mmol CAAE with a final yield of 97.5 % and an enantiometric excess of 99 % was achieved without the addition of cofactor NADPH.

The highly efficient expression of the aspartase gene (L-aspartate ammonia-lyase) in Escherichia coli cells

2015 ◽  
Vol 51 (7) ◽  
pp. 751-756 ◽  
Author(s):  
A. D. Novikov ◽  
D. D. Derbikov ◽  
O. V. Shaposhnikova ◽  
T. A. Gubanova ◽  
S. V. Kameneva ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Highly Efficient ◽  
Escherichia Coli Cells ◽  
Efficient Expression

scholarly journals Ada protein– and sequence context–dependent mutagenesis of alkyl phosphotriester lesions in Escherichia coli cells

2020 ◽  
Vol 295 (26) ◽  
pp. 8775-8783
Author(s):  
Jiabin Wu ◽  
Jun Yuan ◽  
Nathan E. Price ◽  
Yinsheng Wang
Keyword(s):  
Escherichia Coli ◽  
Dna Replication ◽  
Adaptive Response ◽  
Dna Polymerases ◽  
Replication Machinery ◽  
Sequence Context ◽  
E Coli ◽  
Highly Efficient ◽  
Escherichia Coli Cells ◽  
Ada Protein

Alkyl phosphotriester (alkyl-PTE) lesions are frequently induced in DNA and are resistant to repair. Here, we synthesized and characterized methyl (Me)- and n-butyl (nBu)-PTEs in two diastereomeric configurations (Sp and Rp) at six different flanking dinucleotide sites, i.e. XT and TX (X = A, C, or G), and assessed how these lesions impact DNA replication in Escherichia coli cells. When single-stranded vectors contained an Sp-Me-PTE in the sequence contexts of 5′-AT-3′, 5′-CT-3′, or 5′-GT-3′, DNA replication was highly efficient and the replication products for all three sequence contexts contained 85–90% AT and 5–10% TG. Thus, the replication outcome was largely independent of the identity of the 5′ nucleotide adjacent to an Sp-Me-PTE. Furthermore, replication across these lesions was not dependent on the activities of DNA polymerases II, IV, or V; Ada, a protein involved in adaptive response and repair of Sp-Me-PTE in E. coli, however, was essential for the generation of the mutagenic products. Additionally, the Rp diastereomer of Me-PTEs at XT sites and both diastereomers of Me-PTEs at TX sites exhibited error-free replication bypass. Moreover, Sp-nBu-PTEs at XT sites did not strongly impede DNA replication, and other nBu-PTEs displayed moderate blockage effects, with none of them being mutagenic. Taken together, these findings provide in-depth understanding of how alkyl-PTE lesions are recognized by the DNA replication machinery in prokaryotic cells and reveal that Ada contributes to mutagenesis of Sp-Me-PTEs in E. coli.

scholarly journals Synthesis of Chiral Cyanohydrins by Recombinant Escherichia coli Cells in a Micro-Aqueous Reaction System

2012 ◽  
Vol 78 (14) ◽  
pp. 5025-5027 ◽  
Author(s):  
Kathrin Emmi Scholz ◽  
Daniel Okrob ◽  
Benita Kopka ◽  
Alexander Grünberger ◽  
Martina Pohl ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Arabidopsis Thaliana ◽  
Fluorescent Protein ◽  
Reaction System ◽  
Recombinant Escherichia Coli ◽  
Hydroxynitrile Lyase ◽  
Content Type ◽  
Escherichia Coli Cells

ABSTRACTSynthesis of chiral cyanohydrins is performed in a monophasic micro-aqueous reaction system using whole recombinantEscherichia colicells expressing theArabidopsis thalianahydroxynitrile lyase (AtHNL). Microscopy studies employing a fusion ofAtHNL with a flavin-based fluorescent protein (FbFP) reveal that the cells remain intact in the reaction system.

Highly efficient conversion of microcrystalline cellulose to 5-hydroxymethyl furfural in a homogeneous reaction system

RSC Advances ◽  
2016 ◽  
Vol 6 (26) ◽  
pp. 21347-21351 ◽  
Author(s):  
Yan-Ru Zhang ◽  
Nan Li ◽  
Ming-Fei Li ◽  
Yong-Ming Fan
Keyword(s):  
Microcrystalline Cellulose ◽  
Zinc Chloride ◽  
Reaction System ◽  
Homogeneous Reaction ◽  
Hydroxymethyl Furfural ◽  
Efficient Conversion ◽  
Highly Efficient ◽  
System P ◽  
Chloride Hydrate

A homogenous reaction system with zinc chloride hydrate was explored for the synthesis of 5-HMF from MCC.

scholarly journals Production of a Doubly Chiral Compound, (4R,6R)-4-Hydroxy-2,2,6-Trimethylcyclohexanone, by Two-Step Enzymatic Asymmetric Reduction

2003 ◽  
Vol 69 (2) ◽  
pp. 933-937 ◽  
Author(s):  
Masaru Wada ◽  
Ayumi Yoshizumi ◽  
Yumiko Noda ◽  
Michihiko Kataoka ◽  
Sakayu Shimizu ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Saccharomyces Cerevisiae ◽  
Double Bond ◽  
Enzymatic Synthesis ◽  
Asymmetric Reduction ◽  
Glucose Dehydrogenase ◽  
Enantiomeric Excess ◽  
Old Yellow Enzyme ◽  
Chiral Compound ◽  
E Coli

ABSTRACT A practical enzymatic synthesis of a doubly chiral key compound, (4R,6R)-4-hydroxy-2,2,6-trimethylcyclohexanone, starting from the readily available 2,6,6-trimethyl-2-cyclohexen-1,4-dione is described. Chirality is first introduced at the C-6 position by a stereoselective enzymatic hydrogenation of the double bond using old yellow enzyme 2 of Saccharomyces cerevisiae, expressed in Escherichia coli, as a biocatalyst. Thereafter, the carbonyl group at the C-4 position is reduced selectively and stereospecifically by levodione reductase of Corynebacterium aquaticum M-13, expressed in E. coli, to the corresponding alcohol. Commercially available glucose dehydrogenase was also used for cofactor regeneration in both steps. Using this two-step enzymatic asymmetric reduction system, 9.5 mg of (4R,6R)-4-hydroxy-2,2,6-trimethylcyclohexanone/ml was produced almost stoichiometrically, with 94% enantiomeric excess in the presence of glucose, NAD+, and glucose dehydrogenase. To our knowledge, this is the first report of the application of S. cerevisiae old yellow enzyme for the production of a useful compound.

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