利用门多萨假单胞菌NK.docVIP

利用NK-01合成中长链聚羟基脂肪酸酯(PHAMCL)陶 剑1郑承纲1张 斌1 1 王淑芳2 宋存江1, 2南开大学生命科学学院天津 300071(2. 南开大学生命科学学院生物活性材料教育部重点实验室 天津 300071从土壤中分离到株能够合成中长链聚羟基脂肪酸酯(PHAMCL)的细菌, 经分类鉴定命名为Pseudomonas mendocina NK-01。对于该菌株在氮源限制的情况下, 利用葡萄糖合成的PHAMCL进行了结构分析。同时, 获得了该菌株PHA合成酶基因phaC1, 利用大肠杆菌表达载体pBluescript SK构建了重组载体pBSphaC1, 并在E.coli JM109成功表达。以所构建的工程菌株进行了发酵合成。中长链聚羟基脂肪酸酯, 门多萨假单胞菌, PHA合成酶Synthesis of Medium-chain-length Polyhydroxyalkanoate (PHAMCL) by Pseudomonas mendocina NK-01 TAO Jian1,2 ZHENG Cheng-Gang1 ZHANG Bin1 GUO Wen-Bin1 WANG Shu-Fang2 SONG Cun-Jiang1,2* (1. The Key Laboratory of Molecular Microbiology and Technology, Ministry of Education, Nankai University, Tianjin 300071) (2. The Key Laboratory of Bioactive Materials, Ministry of Education, Nankai University, Tianjin 300071) Abstract: A strain which can synthesize medium-chain-length polyhydroxyalkanoate (PHAMCL) was isolated from soil. It was named as Pseudomonas mendocina NK-01 through classification and identification. The PHAMCL was synthesized by P. mendocina NK-01 from glucose in the condition of limited nitrogen source. The structure of it was characterized. In order to construct a recombined plasmid pBSphaC1, an expressing plasmid of E.coli, after phaC1 gene encoding PHA synthase was cloned from Pseudomonas mendocina NK-01 by PCR amplification, it was linked into pBluescript SK?. PhaC1 gene was expressed successfully in E.coli JM109. Fermentation synthesis was also investigated by recombined E.coli. Keywords: Medium-chain-length polyhydroxyalkanoate (PHAMCL), Pseudomonas mendocina, PHA synthase 聚羟基脂肪酸酯(polyhydroxyalkanoate, PHA) 是微生物在失去营养平衡的条件下, 即碳源丰富氮或磷缺失在体内积累的细胞碳源或能源存颗粒, 环境营养条件恢复时这些颗粒可被微生物分解利用。这些高分子聚合物颗粒就是PHA。据报道已有300余种微生物能够合成和积累PHA[1]。由于这类物质具有生物降解性、生物相容性、压电性等特征, 已成为生物材料的研究热点。 PHA含有不同长度碳链的单体侧链基团, 根据单体碳原子数的不同, PHA被划分为两个大类, 即：由35个碳原子的单体组成短链PHA (Short-chain- length Polyhydroxyalkanoate, PHASCL)和由618个碳原子的单体组成中长链PHA (Medium-chain-length Polyhydroxyalkanoate, PHAMCL)[2]。 PHAMCL往往以多种单体组成, 使得该类聚合物拥有了独特的高分子性能。基于PHAMCL的生物降解性