摘要
目的以深海链霉菌Streptomyces somaliensis SCSIO ZH66为研究对象,通过阻断其色氨酸分解代谢途径,探究代谢产物与生长的变化。方法通过Blastp分析寻找S.somaliensis SCSIO ZH66基因组上编码色氨酸双加氧酶基因tdo(ORF0630和ORF6017),在前期阻断ORF0630的基础上采用PCR-targeting的策略阻断ORF6017,通过HPLC检测发酵产物的变化,并观察用不同培养基培养时突变株与野生株生长的差别。结果与野生株相比,突变株ZH66Δtdo不产生antimycins,但无其他次级代谢产物的变化。与此同时,ZH66Δtdo在MS平板上开始产生气生菌丝与孢子的时间均提前了12h,在ISP-2液体培养基中生长对数期开始时间同样提前12h。结论链霉菌S.somaliensis SCSIO ZH66中色氨酸分解代谢途径的阻断未促进其他可能以色氨酸为前体次级代谢产物的合成,而是促进了菌株的生长,为其他链霉菌中色氨酸分解代谢调控提供了借鉴。
Objective To investigate the change of secondary metabolism as well as growth after blocking the tryptophan catabolic pathway in deep-sea-derived Streptomyces somaliensis SCSIO ZH66.Methods Tryptophan-2,3-dioxgenase encoding genes tdo(ORF0630 and ORF6017)were identified from the genome of S.somaliensis SCSIO ZH66 by Blastp search.ORF6017 was knocked out using PCR-targeting strategy based on previously constructed ORF0630 gene inactivation mutant,resulting in ZH66Δtdo.The fermentation products of ZH66Δtdo and wild type strains were analyzed by HPLC,and their growth was observed after cultivation on plates and in liquid media.Results HPLC analysis suggested production of antimycins was abolished in ZH66Δtdo,but no other difference was observed in comparison with the wild type strain.However,formation of the aerial hyphae on MS medium as well as the start of the exponential phase in ISP-2 liquid medium of ZH66Δtdo were both 12 hearlier than that of the wild type strain.Conclusion The disruption of tryptophan catabolic pathway in S.somaliensis SCSIO ZH66 didn’t cause production of tryptophan-derived secondary metabolites,but accelerated the growth of the mutant strain.These results provided a reference for the researches of tryptophan catabolism regulation in other Streptomyces strains.
作者
徐明远
肖菲
李花月
鞠建华
李文利
XU Ming-yuan;XIAO Fei;LI Hua-yue;JU Jian-hua;LI Wen-li(Key Laboratory of Marine Drugs,Ministry Education of China,School of Medicine and Pharmacy, Ocean University of China,Qingdao 266003,China;CAS Key Laboratory of Marine Bio-resourses Sustainable Utilization,Guangdong key Laboratory of Marine Materia Medica,South China Sea Institute of Oceanology, Chinese Academy of Sciences,Guangzhou 510301,China)
出处
《中国海洋药物》
CAS
CSCD
2018年第6期8-14,共7页
Chinese Journal of Marine Drugs
基金
国家自然科学基金项目(41506157,U1706206)资助
关键词
深海链霉菌
色氨酸分解代谢
色氨酸双加氧酶
次级代谢
deep-sea-derived Streptomyces
tryptophan catabolism
tryptophan-2,3-dioxgenase
secondary metabolism
