Breeding High-yield L-isoleucine Escherichia coli by ARTP Mutagenesis

doi:10.13560/j.cnki.biotech.bull.1985.2021-0272

Abstract

Abstract:

This work aims to breed high-yield L-isoleucine-producing Escherichia coli. Using E. coli K12（Met^-）as the original strain,regular temperature and atmospheric pressure plasma（ARTP）was for mutagenesis,combined with microbial high-throughput droplet culture system（MMC）was for screening,with α-aminobutane acid（α-AB）resistance was used as a selection marker to obtain a mutagenic E. coli NXU12 producing high yield of L-isoleucine and its genetic stability was studied. The results showed that after the original strain K12（Met^-）was subjected to ARTP mutagenesis for 180 s,a strain of high-yield L-isoleucine mutagenesis,E. coli NXU12,was selected through single-factor multi-level experiments,adaptive evolution experiments,fermentation culture re-screening,and genetic stability verification. After 40 h of fermentation and culture,the L-isoleucine yield of this strain reached 17.462 8 g/L,which was 61.23% higher than that by original strain 10.830 9 g/L,and the genetic traits were stable.

Key words: Escherichia coli, ARTP, MMC, α-AB, resistance marker, L-isoleucine

LI Xiao-fang, LIU Hui-yan, PAN Lin, AI Zhi-yu, LI Yi-ming, ZHANG Heng, FANG Hai-tian. Breeding High-yield L-isoleucine Escherichia coli by ARTP Mutagenesis[J]. Biotechnology Bulletin, 2022, 38(1): 150-156.

Figures/Tables 8

Fig. 1 Lethality curve of Escherichia coli ARTP mutagenesis

Fig. 2 Single-factor multi-level curve of E. coli NXU12

Fig. 3 Adaptive evolution curve of E. coli NXU12

Fig. 4 Sequencing results of E. coli K12 and NXU12

Fig. 6 E. coli K12 and NXU12 L-isoleucine concentration

Fig. 5 Sugar consumption curve of E. coli K12 and NXU12

Fig. 7 Growth curve of E. coli NXU12 after 10 generations

Fig. 8 L-isoleucine concentration of E. coli NXU12 for 10 generations

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