title: Genome Shuffling and Gentamicin-Resistance to Improve ε-Poly-L-Lysine Productivity of Streptomyces albulus W-156.
abstract: Genome shuffling has been a recently effective method for screening the desirable phenotypes of industrial strains. Here, we combined genome shuffling and gentamicin resistance to improve the production of ε-poly-L-lysine in Streptomyces albulus W-156. Five starting mutants with higher ε-poly-L-lysine (ε-PL) productivities were firstly obtained by atmospheric and room temperature plasma (ARTP) mutagenesis. After three rounds of genome shuffling with increasing concentration of gentamicin for selection, S. albulus AG3-28, was finally got with a production of 3.43 g/L in shaking flask. In a 5-L fermenter, AG3-28 exhibited a higher ε-PL productivity (56.5 g/L) than the initial strain W-156 (37.5 g/L). Key enzyme activities in primary and secondary metabolic pathways were analyzed, and the transcription levels of hrdD and pls were determined by quantitative real time-polymerase chain reaction (qRT-PCR). Increase of key enzyme activities and the upregulation of the gene transcriptional levels demonstrated that ε-PL synthetic pathway in AG3-28 was obviously strengthened, which might be responsible for the high productivity. Moreover, hyper-yield strain AG3-28 was found to produce a slightly lower ε-PL polymerization degree than the parent strain. Amplified fragment length polymorphism (AFLP) analysis reflects the genetic diversity among the derivates after genome shuffling.
authors.affiliation: The Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, 1800 Lihu Road, Wuxi, 214122, Jiangsu, China.
authors.forename: Liang
authors.initials: L
authors.lastname: Wang
authors.affiliation: The Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, 1800 Lihu Road, Wuxi, 214122, Jiangsu, China. chenxs@jiangnan.edu.cn.
authors.forename: Xusheng
authors.initials: X
authors.lastname: Chen
authors.affiliation: The Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, 1800 Lihu Road, Wuxi, 214122, Jiangsu, China.
authors.forename: Guangyao
authors.initials: G
authors.lastname: Wu
authors.affiliation: The Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, 1800 Lihu Road, Wuxi, 214122, Jiangsu, China.
authors.forename: Xin
authors.initials: X
authors.lastname: Zeng
authors.affiliation: The Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, 1800 Lihu Road, Wuxi, 214122, Jiangsu, China.
authors.forename: Xidong
authors.initials: X
authors.lastname: Ren
authors.affiliation: Xiwang Group Company Limited, Zouping, 256209, China.
authors.forename: Shu
authors.initials: S
authors.lastname: Li
authors.affiliation: The Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, 1800 Lihu Road, Wuxi, 214122, Jiangsu, China.
authors.forename: Lei
authors.initials: L
authors.lastname: Tang
authors.affiliation: The Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, 1800 Lihu Road, Wuxi, 214122, Jiangsu, China. maozg@jiangnan.edu.cn.
authors.forename: Zhonggui
authors.initials: Z
authors.lastname: Mao
mesh_terms: D054458:Amplified Fragment Length Polymorphism Analysis; D061249:Batch Cell Culture Techniques; D019149:Bioreactors / Q000382:microbiology; D032621:DNA Shuffling; D004352:Drug Resistance, Microbial / Q000187:drug effects / Q000235:genetics*; D005285:Fermentation / Q000187:drug effects; D015964:Gene Expression Regulation, Bacterial / Q000187:drug effects; D005798:Genes, Bacterial; D016680:Genome, Bacterial; D005839:Gentamicins / Q000494:pharmacology*; D016296:Mutagenesis; D009154:Mutation / Q000235:genetics; D058626:Plasma Gases / Q000737:chemistry; D011107:Polylysine / Q000096:biosynthesis* / Q000737:chemistry; D058105:Polymerization; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D011995:Recombination, Genetic / Q000235:genetics; D013302:Streptomyces / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism*; D013696:Temperature; D013997:Time Factors; D014158:Transcription, Genetic / Q000187:drug effects
chemical_list: D005839:Gentamicins; D058626:Plasma Gases; D012333:RNA, Messenger; D011107:Polylysine
keywords: AFLP; Genome shuffling; Gentamicin resistance mutation; Key enzyme activities; Polymerization; qRT-PCR; ε-Poly-L-lysine