Optimizing the DO‐stat protocol for enhanced production of thermostable pullulanase in Escherichia coli by using oxygen control strategies

2020 ◽  
Vol 44 (5) ◽  
Author(s):  
Lei Chi ◽  
Jiajia Wei ◽  
Junchao Hou ◽  
Jingyu Wang ◽  
Xiaolong Hu ◽  
...  
2005 ◽  
Vol 71 (12) ◽  
pp. 7880-7887 ◽  
Author(s):  
Sang Jun Lee ◽  
Dong-Yup Lee ◽  
Tae Yong Kim ◽  
Byung Hun Kim ◽  
Jinwon Lee ◽  
...  

ABSTRACT Comparative analysis of the genomes of mixed-acid-fermenting Escherichia coli and succinic acid-overproducing Mannheimia succiniciproducens was carried out to identify candidate genes to be manipulated for overproducing succinic acid in E. coli. This resulted in the identification of five genes or operons, including ptsG, pykF, sdhA, mqo, and aceBA, which may drive metabolic fluxes away from succinic acid formation in the central metabolic pathway of E. coli. However, combinatorial disruption of these rationally selected genes did not allow enhanced succinic acid production in E. coli. Therefore, in silico metabolic analysis based on linear programming was carried out to evaluate the correlation between the maximum biomass and succinic acid production for various combinatorial knockout strains. This in silico analysis predicted that disrupting the genes for three pyruvate forming enzymes, ptsG, pykF, and pykA, allows enhanced succinic acid production. Indeed, this triple mutation increased the succinic acid production by more than sevenfold and the ratio of succinic acid to fermentation products by ninefold. It could be concluded that reducing the metabolic flux to pyruvate is crucial to achieve efficient succinic acid production in E. coli. These results suggest that the comparative genome analysis combined with in silico metabolic analysis can be an efficient way of developing strategies for strain improvement.


2019 ◽  
Vol 7 (11) ◽  
pp. 552 ◽  
Author(s):  
Ziaur Rahman ◽  
Bong Hyun Sung ◽  
Javed Nawab ◽  
Muhammad Faisal Siddiqui ◽  
Abid Ali ◽  
...  

Biodiesel, or fatty acid ethyl ester (FAEE), is an environmentally safe, next-generation biofuel. Conventionally, FAEE is produced by the conversion of oil/fats, obtained from plants, animals, and microorganisms, by transesterification. Recently, metabolic engineering of bacteria for ready-to-use biodiesel was developed. In Escherichia coli, it is produced by fatty acyl-carrier proteins and ethanol, with the help of thioesterase (TesB) and wax synthase (WS) enzymes. One of the foremost barriers in microbial FAEE production is the feedback inhibition of the fatty acid (FA) operon (fabHDG). Here, we studied the effect of biodiesel biosynthesis in E. coli with an engineered fabHDG operon. With a basic FAEE producing BD1 strain harboring tes and ws genes, biodiesel of 32 mg/L were produced. Optimal FAEE biosynthesis was achieved in the BD2 strain that carries an overexpressed operon (fabH, fabD, and fabG genes) and achieved up to 1291 mg/L of biodiesel, a 40-fold rise compared to the BD1 strain. The composition of FAEE obtained from the BD2 strain was 65% (C10:C2, decanoic acid ethyl ester) and 35% (C12:C2, dodecanoic acid ethyl ester). Our findings indicate that overexpression of the native FA operon, along with FAEE biosynthesis enzymes, improved biodiesel biosynthesis in E. coli.


2012 ◽  
Vol 56 (11) ◽  
pp. 5575-5580 ◽  
Author(s):  
Jennifer H. Han ◽  
Kei Kasahara ◽  
Paul H. Edelstein ◽  
Warren B. Bilker ◽  
Ebbing Lautenbach

ABSTRACTThere has been a significant increase in the prevalence ofEnterobacteriaceaethat produce CTX-M-type extended-spectrum β-lactamases. The objective of this study was to evaluate risk factors for infection or colonization with CTX-M-positiveEscherichia coli. A case-control study was conducted within a university system from 1 January 2007 to 31 December 2008. All patients with clinical cultures withE. colidemonstrating resistance to extended-spectrum cephalosporins were included. Case patients were designated as those with cultures positive for CTX-M-positiveE. coli, and control patients were designated as those with non-CTX-M-producingE. coli. Multivariable logistic regression analyses were performed to evaluate risk factors for CTX-M-positive isolates. A total of 83 (56.8%) of a total of 146 patients had cultures with CTX-M-positiveE. coli. On multivariable analyses, there was a significant association between infection or colonization with CTX-M-type β-lactamase-positiveE. coliand receipt of piperacillin-tazobactam in the 30 days prior to the culture date (odds ratio [OR], 7.36; 95% confidence interval [CI], 1.61 to 33.8;P= 0.01) and a urinary culture source (OR, 0.36; 95% CI, 0.17 to 0.77;P= 0.008). The rates of resistance to fluoroquinolones were significantly higher in isolates from case patients than in isolates from control patients (90.4 and 50.8%, respectively;P< 0.001). We found that nonurinary sources of clinical cultures and the recent use of piperacillin-tazobactam conferred an increased risk of colonization or infection with CTX-M-positiveE. coli. Future studies will need to focus on outcomes associated with infections due to CTX-M-positiveE. coli, as well as infection control strategies to limit the spread of these increasingly common organisms.


RSC Advances ◽  
2018 ◽  
Vol 8 (53) ◽  
pp. 30512-30519 ◽  
Author(s):  
Jian-Xiu Li ◽  
Yan-Yan Huang ◽  
Xian-Rui Chen ◽  
Qi-Shi Du ◽  
Jian-Zong Meng ◽  
...  

Enhanced production of optical (S)-acetoin by a recombinant Escherichia coli whole-cell biocatalyst with NADH regeneration systems.


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