链霉菌纤维素酶基因的异源表达及产抗生素链霉菌的分子育种

K. Kasuga, Y. Chida, Ayaka Sabanai, Shunichiro Kyono, Kumiko Tomotsune, J. Ishikawa, H. Ikeda, Ikuo Kojima
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引用次数: 0

摘要

从土壤中分离出来,用于从纤维素生物质中产生抗生素的链霉菌的遗传育种。初步的基因组测序显示,编码9种纤维素酶和1种木糖葡聚糖酶的基因分散在染色体上。将这些基因分离并重组到链霉菌pTYM19染色体整合载体上,构建链霉菌宿主菌株的纤维素酶表达质粒pBOM51和pBOM66。将pBOM66上的纤维素酶基因簇进一步导入另一链霉菌整合载体pTYM18中,得到pBOM67。为了研究纤维素酶的分泌和抗生素的产生,将质粒导入产生阿维菌素和寡霉素的阿维菌链霉菌K139。通过携带pbom66的变压器观察到对滤纸纤维素的显著降解活性。只有在含有葡萄糖的培养基中培养,而不是在含有微晶纤维素作为碳源的培养基中培养时,用pBOM67转化的菌株才能产生寡霉素。因此,利用产生抗生素的链霉菌从纤维素生物质中生产抗生素似乎是不可行的,即使用链霉菌来源的纤维素酶基因转化也是如此。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Heterologous Expression of Streptomyces Cellulase Genes for the Molecular Breeding of Antibiotic Producing Streptomycetes from Cellulosic Biomass
was isolated from soil for genetic breeding of streptomycetes that produce antibiotics from cellulosic biomass. Draft genome sequencing revealed putative genes encoding nine cellulases and one xyloglucanase dispersed on the chromosome. All these genes were isolated and rearranged on a chromosome-integration vector for streptomycetes pTYM19 to construct cellulase-expression plasmids pBOM51 and pBOM66 for streptomycete host strains. The cellulase gene cluster on pBOM66 was further introduced into pTYM18, another Streptomyces integration vector, to yield pBOM67. To investigate cellulase secretion and antibiotic production, the resulting plasmids were introduced into Streptomyces avermitilis K139, the producer of avermectin and oligomycin. Remarkable cellulose-degrading activity for fi lter paper was observed by pBOM66-carrying transformants. Oligomycin alone was produced by the transformed strain with pBOM67 only when cultivated in a medium containing glucose: not in a medium containing microcrystalline cellulose as a carbon source. Using antibiotic-producing streptomycetes for antibiotic production from cellulosic biomass therefore appears to be infeasible, even when transformed with cellulase genes of Streptomyces origin.
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