摘要
利用Genome shuffling技术对1株融合重组菌Streptomyces sp.Feel-1产ε-聚赖氨酸(ε-PL)进行了育种研究。首先对Feel-1进行了甲基磺酸乙酯(EMS)诱变,以甘氨酸(Gly)和S-2-氨乙基-L-半胱氨酸(AEC)作为抗性指标进行筛选,得到4株正向突变株,ε-PL摇瓶产量分别为1.78 g/L、1.89 g/L、1.85 g/L、1.86 g/L,比原始菌株Feel-1(1.60 g/L)分别提高了11.3%、18.1%、15.6%、16.3%;然后将这4株产量提高株作为出发菌库,采用双亲灭活法进行了3轮递推式原生质体融合(Genome shuffling),最终得到了3株产量大幅提高的重组菌,摇瓶产量分别为2.42 g/L、2.35 g/L和2.37 g/L,比原始菌株分别提高了51.3%、46.9%和48.1%;选择其中1株(G3-67)进行补料分批发酵,192 h时ε-PL产量达到32.2 g/L。
ABST cing e supple were o RACT Genome shuffling was adopted to breed a recomb -poly-L-lysine （e-PL） in this research. First Feel -1 was mented with Gly and AEC （S-2-ammonia ethyl-L-homocys btained in shak spectively, which was these four positive mu inant strain named Streptomyces sp. Feel-1 for produmutated by mutagen EMS and spread on agar plate teine） as resistance markers. Four improved mutants e-flask fermentation with e-PL production as 1.78 g/L, 1.89 g/L, 1.85 g/L, 1.86 g/L, reincreased by 11.3% , 18.1% , 15.6% and 16.3% compared with Feel-1 （1.6 g/L ）. Then tants were used as starting population and subjected for three rounds of genome shuffling by fu- sing the inactivated protoplasts. Finally three recombinan 2.42 g/L, 2.35 g/L and 2.37 g/L, respectively, which tively. Fed-batch fermentation by one of the recombinants to 35 g/L,
出处
《食品与发酵工业》
CAS
CSCD
北大核心
2012年第8期36-41,共6页
Food and Fermentation Industries
基金
无锡市科技支撑计划项目(CYE21N1107)
关键词
Ε-聚赖氨酸
Genomeshuffling
原生质体融合
发酵
which was the highest e-poly-L-lysine, geno reported level in China me shuffling, protoplast ts were proved to give a high yield of e-PL production as was over Feel-1 by 51.3% , 46.9% and 48.1% , respec- G3-67 was carried out and the e-PL production accumulated at present. Fusion, fermentation
