Effect of Tween 40 and DtsR1 on l-arginine overproduction in Corynebacterium crenatum
| Autor: | Xue-Lan Chen, Fang Wan, Bin Zhang, Xiong Yonghua, Jincong Chen, Chen Minliang |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2015 |
| Předmět: |
l-Arginine
Corynebacterium crenatum Arginine Mutant Corynebacterium Glutamic Acid Polysorbates Bioengineering Biology Applied Microbiology and Biotechnology Tween 40 Corynebacterium glutamicum Microbiology Gene Knockout Techniques Polyoxyethylene sorbitan monopalmitate DtsR1 Overproduction NADP+/NADPH Research Glutamic acid Gene Expression Regulation Bacterial biology.organism_classification Biochemistry α-Oxoglutarate dehydrogenase complex Genes Bacterial bacteria Biotechnology |
| Zdroj: | Microbial Cell Factories |
| ISSN: | 1475-2859 |
| Popis: | Background l-Glutamate is an important precursor in the l-arginine (l-Arg) biosynthetic pathway. Various methods, including polyoxyethylene sorbitan monopalmitate (Tween 40) addition and dtsR1 disruption, have been widely used to induce l-glutamate overproduction in Corynebacterium glutamicum. In this study, a novel strategy for l-Arg overproduction through Tween 40 trigger and ΔdtsR1 mutant were proposed in Corynebacterium crenatum. Results Corynebacterium crenatum mutant (CCM01) was selected as a host strain, whose argR was lethal via mutagenesis screening, the proB gene was knocked out, and argB was replaced by argB M4 (E19R, H26E, D311R, and D312R) to release l-Arg feedback resistance. After Tween 40 trigger in the logarithmic period, l-Arg production increased from 15.22 to 17.73 g/L in CCM01 strain. When NCgl1221 and dtsR1 disruption (CCM03), l-Arg production drastically increased to 27.45 g/L and then further to 29.97 g/L after Tween 40 trigger. Moreover, the specific activity of α-oxoglutarate dehydrogenase complex (ODHC) decreased, whereas the regeneration of NADP+/NADPH significantly increased after dtsR1 disruption and Tween 40 trigger. Results of real-time PCR showed that the transcriptional levels of odhA, sucB, and lpdA (encoding three subunits of the ODHC complex) were downregulated after Tween 40 trigger or dtsR1 disruption. By contrast, zwf transcription (encoding glucose-6-phosphate dehydrogenase) showed no significant difference among CCM01, CCM02 (ΔNCgl1221), and CCM03 (ΔNCgl1221ΔdtsR1) strains without Tween 40 trigger but evidently increased by 5.50 folds after Tween 40 trigger. Conclusion A novel strategy for l-Arg overproduction by dtsR1 disruption and Tween 40 trigger in C. crenatum was reported. Tween 40 addition exhibited a bifunctional mechanism for l-Arg overproduction, including reduced ODHC activity and enhanced NADPH pools accumulation by downregulated dtsR1 expression and upregulated zwf expression, respectively. |
| Databáze: | OpenAIRE |
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