Genome shuffling and ribosome engineering of Streptomyces actuosus for high-yield nosiheptide production.
Nosiheptide is one of the EU-approved sulfur-containing peptides in feed industry to inhibit the growth of the majority of Gram-positive bacteria. The main purpose of this study is directed to breed the high nosiheptide-producers by genome shuffling and ribosome engineering in Streptomyces actuosus AW7. The starting population for shuffling was generated by combining (60)Coγ-irradiation with LiCl mutagenesis treatments on the spores. After four rounds of protoplast fusion exposed to streptomycin as adaptive pressure, a high-yield recombinant strain D92 was obtained. In a 10-L fermenter, nosiheptide production reached 1.54 g/L which was 9.20-fold compared to that of the parental strain. Hyphae development, metabolic process, and ribosomal protein S12 sequence were investigated to characterize the differentiation among the recombinants. Several mutations in S12 were believed to be responsible to streptomycin resistance in the tested strain. The results demonstrated that the combination of genome shuffling and ribosome engineering is an efficient approach to breed high-yield industrial strains.
Department of Biological Engineering, Zhejiang Gongshang University, Hangzhou, 310035, China.
Qingling
Q
Wang
Dong
D
Zhang
Yudong
Y
Li
Fuming
F
Zhang
Cao
C
Wang
Xinle
X
Liang
D000821:Animal Feed; D000900:Anti-Bacterial Agents / Q000096:biosynthesis*; D032621:DNA Shuffling / Q000379:methods; D005285:Fermentation; D005503:Food Additives; D016680:Genome, Bacterial; D007218:Industrial Microbiology; D016296:Mutagenesis; D012270:Ribosomes / Q000235:genetics / Q000378:metabolism; D013302:Streptomyces / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism*; D013844:Thiazoles / Q000378:metabolism
D000900:Anti-Bacterial Agents; D005503:Food Additives; D013844:Thiazoles; C014134:nosiheptide