Breeding L-arginine-producing strains by a novel mutagenesis method: Atmospheric and room temperature plasma (ARTP)

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.

Elevated H2AX Phosphorylation Observed with kINPen Plasma Treatment Is Not Caused by ROS-Mediated DNA Damage but Is the Consequence of Apoptosis

“…Its efficacy seemed greater than that of several conventional methods, e.g., chemicals and UV radiation [82], and ARTP can also be combined with such methods [83]. As result, ARTP was found to increase production of, for instance, α -ketoglutaric acid [84], biofuel [85], polysaccharides [86, 87], arachidonic acid [88], erythritol [89], l-arginine [90], alkaline α -amylase [91, 92], d-lactic acid [93], lycopene [94], and carotenoids and lipids [95] in different types of microorganisms. However, it is important to mention that these plasma sources are not intended for medical applications.…”

Astaxanthin overproduction in yeast by strain engineering and new gene target uncovering

“…New molecular targets identified in mutagenesis are just valuable feedback to further rational design. Among physical and chemical methods for mutagenesis [ 15 ], an atmospheric and room temperature plasma (ARTP) method has been applied for mutagenesis of various species to obtain targeted biological features [ 16 , 17 ]. The ARTP mutation system can induce diverse breakage in plasmid DNA and oligonucleotides with variation of plasma dosage [ 18 ].…”

Strategies for directed and adapted evolution as part of microbial strain engineering

“…70 For example, Zhou et al 71,72 successfully used ARTP to mutate and evolve Candida glabrata and Yarrowia lipolytica for improved pyruvate and alpha-ketoglutaric acid production, respectively. In addition, a variety of additional chemical overproduction and tolerance phenotypes have been achieved using ARTP mutagenesis followed by screening and selection including ethanol-tolerance, 73 lycopene production, 74 enzyme production, 75,76 cell growth and succinic acid production, 77 butanol production, 78 amino acid production, 79 and lipid productivity. 80,81 The coupling of these approaches with other selections (including adaptive laboratory evolution) has proven to be a successful method as demonstrated by Ma et al 77 by coupling ARTP and ALE to improve anaerobic cell growth and succinic acid production in E. coli 3.12-and 2.5-fold, respectively.…”