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一株芘降解菌B2的降解条件优化及降解基因

Keywords: 假单胞菌属,芘代谢,正交实验设计,邻苯二酚-2,3-双加氧酶

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Abstract:

芳香烃降解菌是石油污染土壤修复的主要生物资源。采用芘平板升华法对克拉玛依原油污染土壤样品进行驯化培养,分离得到一株芘降解菌B2,经16SrDNA基因序列比对及系统发育进化分析表明,该菌株为假单胞菌属(Pseudomonas)。采用正交设计方法优化菌株B2对高分子量多环芳烃芘的降解条件,并构建多元非线性模型预测菌株B2对芘的最佳降解条件,结果表明:在接种量OD660nm为0.60、降解温度为40℃、降解时间为6.0d时,预测菌株B2对芘的降解最大达到38.214mg/L,实际测得最大降解量为37.906mg/L,预测准确率为99.19%。运用PCR技术克隆B2的邻苯二酚-2,3-双加氧酶基因(B2C23O)(I.2.A亚家族),核酸序列分析表明,该基因全长880bp,具有一个完整的开放阅读框,编码246个氨基酸,与已报道的i>PseudomonasputidaW619同源性最高为97%;对B2C23O基因编码氨基酸序列进行分析,发现其具有邻位断裂双加氧酶模式结构,推测菌株B2通过邻位裂解途径降解芘代谢中间产物邻苯二酚。

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