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Updated: Apr 1, 2026

Mutagenesis and Functional Selection Protocols for Directed Evolution of Proteins in E. coli
Published on: March 16, 2011
Breeding L-arginine-producing strains by a novel mutagenesis method: Atmospheric and room temperature plasma (ARTP)
Gong Cheng1, Jianzhong Xu1, Xiuhua Xia2
1a The Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology , Jiangnan University , Wuxi , China.
Abstract:
A plasma jet, driven by an active helium atom supplied with an atmospheric and room temperature plasma (ARTP) biological breeding system, was used as a novel method to breed L-arginine high-yielding strains. A mutant with resistance to L-homoarginine and 8-azaguaine, ARG 3-15 (L-HA(r), 8-AG(r), L-His(-)), was screened after several rounds of screening. The L-arginine production of these mutants was more than that of the original strain, increased by 43.79% for ARG 3-15. Moreover, N-acetyl-L-glutamate synthase activity of these mutants was also increased. After a series of passages, the hereditary properties of these mutants were found to be stable. Interestingly, beet molasses was utilized in a co-feeding fermentation and benefited to increase the productivity by 5.88%. Moreover, the fermentation with 1.0 g/L betaine could produce 9.33% more L-arginine than without betaine. In fed-batch fermentation, C. glutamicum ARG 3-15 began to produce L-arginine at the initial of logarithmic phase, and continuously increased over 24 hr to a final titer of 45.36 ± 0.42 g/L. The L-arginine productivity was 0.571 g/L/hr and the conversion of glucose (α) was 32.4% after 96 hr. These results indicated that C. glutamicum ARG 3-15 is a promising industrial producer.
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