scholarly journals Carbon catabolite repression relaxation in Escherichia coli: global and sugar-specific methods for glucose and secondary sugar co-utilization

2020 ◽  
Vol 30 ◽  
pp. 9-16
Author(s):  
Kevin J Fox ◽  
Kristala LJ Prather
2021 ◽  
Vol 64 (1) ◽  
Author(s):  
Hyeon Jeong Seong ◽  
Yu-Sin Jang

AbstractEscherichia coli has been used as a host to construct the cell factory for biobased production of chemicals from renewable feedstocks. Because galactose is found in marine biomass as a major component, the strategy for galactose utilization in E. coli has been gained more attention. Although galactose and glucose co-fermentation has been reported using the engineered E. coli strain, few reports have covered fermentation supplemented with galactose as a sole carbon source in the mutant lacking the repressor-specific carbon catabolite repression (CCR). Here, we report the effects of the deregulation of the repressor-specific CCR (galR− and galS−) in fermentation supplemented with galactose as a sole carbon source, using the engineered E. coli strains. In the fermentation using the galR− and galS− double mutant (GR2 strain), an increase of rates in sugar consumption and cell growth was observed compared to the parent strain. In the glucose fermentation, wild-type W3110 and its mutant GR2 and GR2PZ (galR−, galS−, pfkA−, and zwf−) consumed sugar at a higher rate than those values obtained from galactose fermentation. However, the GR2P strain (galR−, galS−, and pfkA−) showed no difference between fermentations using glucose and galactose as a sole carbon source. This study provides essential information for galactose fermentation using the CCR-deregulated E. coli strains.


2015 ◽  
Vol 23 (2) ◽  
pp. 99-109 ◽  
Author(s):  
A. Kremling ◽  
J. Geiselmann ◽  
D. Ropers ◽  
H. de Jong

2009 ◽  
Vol 76 (5) ◽  
pp. 1524-1532 ◽  
Author(s):  
Tasha A. Desai ◽  
Christopher V. Rao

ABSTRACT Bacteria such as Escherichia coli will often consume one sugar at a time when fed multiple sugars, in a process known as carbon catabolite repression. The classic example involves glucose and lactose, where E. coli will first consume glucose, and only when it has consumed all of the glucose will it begin to consume lactose. In addition to that of lactose, glucose also represses the consumption of many other sugars, including arabinose and xylose. In this work, we characterized a second hierarchy in E. coli, that between arabinose and xylose. We show that, when grown in a mixture of the two pentoses, E. coli will consume arabinose before it consumes xylose. Consistent with a mechanism involving catabolite repression, the expression of the xylose metabolic genes is repressed in the presence of arabinose. We found that this repression is AraC dependent and involves a mechanism where arabinose-bound AraC binds to the xylose promoters and represses gene expression. Collectively, these results demonstrate that sugar utilization in E. coli involves multiple layers of regulation, where cells will consume first glucose, then arabinose, and finally xylose. These results may be pertinent in the metabolic engineering of E. coli strains capable of producing chemical and biofuels from mixtures of hexose and pentose sugars derived from plant biomass.


2019 ◽  
Vol 81 (1) ◽  
pp. 52-55 ◽  
Author(s):  
Charles E. Deutch

The Bio-Rad pGLO bacterial transformation kit is commonly used to demonstrate this form of genetic exchange, which occurs in bacteria and eukaryotes and which differs fundamentally from transduction and conjugation. The basic experiment leads to the formation of green fluorescent colonies of Escherichia coli and can be extended to illustrate the specificity of the interaction between sugars and the AraC protein, the phenomenon of carbon catabolite repression, the substrate specificity of the β-lactamase encoded by the plasmid, and the role of host restriction/modification systems in the transformation process. pGLO DNA also can be isolated using plasmid mini-prep kits, analyzed with restriction endonucleases, and used to study the conditions for transformation in more detail.


1999 ◽  
Vol 181 (5) ◽  
pp. 1610-1616 ◽  
Author(s):  
Ivo E. Staijen ◽  
Rosanna Marcionelli ◽  
Bernard Witholt

ABSTRACT The alk genes are located on the OCT plasmid ofPseudomonas oleovorans and encode an inducible pathway for the utilization of n-alkanes as carbon and energy sources. We have investigated the influence of alternative carbon sources on the induction of this pathway in P. oleovorans andEscherichia coli alk + recombinants. In doing so, we confirmed earlier reports that induction of alkane hydroxylase activity in pseudomonads is subject to carbon catabolite repression. Specifically, synthesis of the monooxygenase component AlkB is repressed at the transcriptional level. The alk genes have been cloned into plasmid pGEc47, which has a copy number of about 5 to 10 per cell in both E. coli and pseudomonads.Pseudomonas putida GPo12 is a P. oleovoransderivative cured of the OCT plasmid. Upon introduction of pGEc47 in this strain, carbon catabolite repression of alkane hydroxylase activity was reduced significantly. In cultures of recombinant E. coli HB101 and W3110 carrying pGEc47, induction of AlkB and transcription of the alkB gene were no longer subject to carbon catabolite repression. This suggests that carbon catabolite repression of alkane degradation is regulated differently inPseudomonas and in E. coli strains. These results also indicate that P alkBFGHJKL , the P alk promoter, might be useful in attaining high expression levels of heterologous genes in E. coligrown on inexpensive carbon sources which normally trigger carbon catabolite repression of native expression systems in this host.


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