引用本文:	胡斌,韩峰,林育姿,宫倩红,于文功.富含古罗糖醛酸的褐藻多糖生产菌株的构建[J].海洋科学,2008,32(1):14-18.

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富含古罗糖醛酸的褐藻多糖生产菌株的构建
胡斌¹, 韩峰¹, 林育姿¹, 宫倩红¹, 于文功¹
中国海洋大学医药学院教育部海洋药物重点实验室

摘要:

为获得古罗糖醛酸(Guluronate)含量高的细菌胞外褐藻多糖,利用PCR从海洋细菌Pseudomonas sp. QDA中克隆了其甘露糖醛酸C-5差向异构酶基因(algG),连接入质粒pMF 54Km,构建了重组表达载体pMF54 Km-algG。利用三亲接合法将pMF54 Km-algG转入菌株QDA中,获得algG过量表达重组菌株QDA-G。¹H-NMR测定结果表明,QDA-G所产的褐藻多糖中β-D-甘露糖醛酸(M)与它的C-5差向异构体α-L-古罗糖醛酸(G)的比值为0.38,G的质量分数达到74.2%,比野生菌株QDA提高了26.4%。且重组菌株遗传稳定性良好,连续传代20代后,M/G的比值无明显变化。

关键词: 甘露糖醛酸C-5差向异构酶假单胞菌褐藻多糖过量表达

DOI：

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基金项目:国家自然科学基金资助项目( 30371683)

Construction of high-guluronate-containing-alginate producing mutant of Pseudomonas sp.QDA

胡斌,韩峰

Abstract:

Pseudomonas sp. QDA, which was isolated from seawater, has the ability of producing extracellular alginate. In order to obtain the high-guluronate-containing alginate, C-5-mannuronan epimerase gene(algG) of Pseudomonas sp. QDA was cloned by PCR and ligated in to plasmid pMF54Km. The resulting plasmid pMF54Km-algG was transferred to QDA by triparental mating,and an algG over-expressing strain, QDA-G, was obtained. The results of ¹H-NMR showed that the M/G ratio of alginate fragments by QDA-G was 0.38 and the content of guluronate was 74.2%, about 26.4% higher than that of wild strain QDA. The genetic stability of QDA-G was good. No significant changes were found in M/G ratio of alginate fragments of QDA-G after 20 passages.

Key words: mannuronan C-5 epimerases Pseudomonas sp.QDA alginate over-expression