The rebalanced pathway significantly enhances acetoin production by disruption of acetoin reductase gene and moderate-expression of a new water-forming NADH oxidase in Bacillus subtilis

2014 ◽  
Vol 23 ◽  
pp. 34-41 ◽  
Author(s):  
Xian Zhang ◽  
Rongzhen Zhang ◽  
Teng Bao ◽  
Zhiming Rao ◽  
Taowei Yang ◽  
...  
Keyword(s):  
Bacillus Subtilis ◽  
Nadh Oxidase ◽  
Reductase Gene ◽  
Acetoin Production

scholarly journals Design of a programmable biosensor-CRISPRi genetic circuits for dynamic and autonomous dual-control of metabolic flux in Bacillus subtilis

2019 ◽  
Vol 48 (2) ◽  
pp. 996-1009 ◽  
Author(s):  
Yaokang Wu ◽  
Taichi Chen ◽  
Yanfeng Liu ◽  
Rongzhen Tian ◽  
Xueqin Lv ◽  
...  
Keyword(s):  
Bacillus Subtilis ◽  
Metabolic Pathways ◽  
Metabolic Flux ◽  
Target Genes ◽  
Fine Tuning ◽  
Dual Control ◽  
Genetic Circuits ◽  
Dynamic Regulation ◽  
Batch Bioreactor ◽  
Acetoin Production

Abstract Dynamic regulation is an effective strategy for fine-tuning metabolic pathways in order to maximize target product synthesis. However, achieving dynamic and autonomous up- and down-regulation of the metabolic modules of interest simultaneously, still remains a great challenge. In this work, we created an autonomous dual-control (ADC) system, by combining CRISPRi-based NOT gates with novel biosensors of a key metabolite in the pathway of interest. By sensing the levels of the intermediate glucosamine-6-phosphate (GlcN6P) and self-adjusting the expression levels of the target genes accordingly with the GlcN6P biosensor and ADC system enabled feedback circuits, the metabolic flux towards the production of the high value nutraceutical N-acetylglucosamine (GlcNAc) could be balanced and optimized in Bacillus subtilis. As a result, the GlcNAc titer in a 15-l fed-batch bioreactor increased from 59.9 g/l to 97.1 g/l with acetoin production and 81.7 g/l to 131.6 g/l without acetoin production, indicating the robustness and stability of the synthetic circuits in a large bioreactor system. Remarkably, this self-regulatory methodology does not require any external level of control such as the use of inducer molecules or switching fermentation/environmental conditions. Moreover, the proposed programmable genetic circuits may be expanded to engineer other microbial cells and metabolic pathways.

High acetoin production by a newly isolated marine Bacillus subtilis strain with low requirement of oxygen supply

2015 ◽  
Vol 50 (11) ◽  
pp. 1730-1734 ◽  
Author(s):  
Jian-Ying Dai ◽  
Ling Cheng ◽  
Qing-Feng He ◽  
Zhi-Long Xiu
Keyword(s):  
Bacillus Subtilis ◽  
Oxygen Supply ◽  
Subtilis Strain ◽  
Acetoin Production

Various means of integration of the expressible human dihydrofolate reductase gene into the Bacillus subtilis genome

Gene ◽  
1987 ◽  
Vol 57 (2-3) ◽  
pp. 221-227 ◽  
Author(s):  
Alexander A. Prozorov ◽  
Elena U. Poluektova ◽  
Galina V. Savchenko ◽  
Venera Z. Nezametdinova ◽  
Fuat K. Khasanov
Keyword(s):  
Bacillus Subtilis ◽  
Dihydrofolate Reductase ◽  
Reductase Gene ◽  
Human Dihydrofolate Reductase ◽  
Subtilis Genome

Characterization of xylitol 4-dehydrogenase from Erwinia aphidicola and its co-expression with NADH oxidase in Bacillus subtilis

2021 ◽  
Vol 104 ◽  
pp. 92-100
Author(s):  
Mengli Li ◽  
Wenhui Zhu ◽  
Qing Meng ◽  
Ming Miao ◽  
Tao Zhang
Keyword(s):  
Bacillus Subtilis ◽  
Nadh Oxidase

scholarly journals Efficient 2,3-Butanediol/Acetoin Production Using Whole-Cell Biocatalyst with a New Nadh/Nad(+) Regeneration System

Catalysts ◽  
2021 ◽  
Vol 11 (12) ◽  
pp. 1422
Author(s):  
Yaping Wang ◽  
Yanhong Peng ◽  
Xiaoyan Liu ◽  
Ronghua Zhou ◽  
Xianqing Liao ◽  
...  
Keyword(s):  
Formate Dehydrogenase ◽  
Target Genes ◽  
Nadh Oxidase ◽  
Expression System ◽  
Regeneration System ◽  
Nadh Regeneration ◽  
Whole Cell ◽  
Whole Cell Biocatalyst ◽  
Acetoin Production ◽  
Butanediol Dehydrogenase

An auto-inducing expression system was developed that could express target genes in S. marcescens MG1. Using this system, MG1 was constructed as a whole-cell biocatalyst to produce 2,3-butanediol/acetoin. Formate dehydrogenase (FDH) and 2,3-butanediol dehydrogenase were expressed together to build an NADH regeneration system to transform diacetyl to 2,3-butanediol. After fermentation, the extract of recombinant S. marcescens MG1ABC (pETDuet-bdhA-fdh) showed 2,3-BDH activity of 57.8 U/mg and FDH activity of 0.5 U/mg. And 27.95 g/L of 2,3-BD was achieved with a productivity of 4.66 g/Lh using engineered S. marcescens MG1(Pswnb+pETDuet-bdhA-fdh) after 6 h incubation. Next, to produce 2,3-butanediol from acetoin, NADH oxidase and 2,3-butanediol dehydrogenase from Bacillus subtilis were co-expressed to obtain a NAD+ regeneration system. After fermentation, the recombinant strain S. marcescens MG1ABC (pSWNB+pETDuet-bdhA-yodC) showed AR activity of 212.4 U/mg and NOX activity of 150.1 U/mg. We obtained 44.9 g/L of acetoin with a productivity of 3.74 g/Lh using S. marcescens MG1ABC (pSWNB+pETDuet-bdhA-yodC). This work confirmed that S. marcescens could be designed as a whole-cell biocatalyst for 2,3-butanediol and acetoin production.

Engineering genome-reduced Bacillus subtilis for acetoin production from xylose

2017 ◽  
Vol 40 (2) ◽  
pp. 393-398 ◽  
Author(s):  
Panpan Yan ◽  
Yuanqing Wu ◽  
Li Yang ◽  
Zhiwen Wang ◽  
Tao Chen
Keyword(s):  
Bacillus Subtilis ◽  
Acetoin Production
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