响应面法优化芘降解菌的降解条件及降解基因分析

发布时间：2018-02-15 06:08

本文关键词： 芘降解菌邻苯二酚- -双加氧酶响应面法优化　出处：《新疆大学学报(自然科学版)》2017年02期 　论文类型：期刊论文

【摘要】：采用响应面法进行实验设计,选择p H值、温度、天数为影响因素,对芘浓度为50 mg/L无机盐培养基进行wp3-1菌株降解条件优化,得到了芘降解率与3种因素间的非线性回归方程,确定了降解芘的最优工艺条件:在温度为34.15?C,p H值为6.98,降解时间为4.28天时,预测芘的降解率为86.42%,实验验证值为85.70%,结果显示,建立的模型具有较高的精度.采用PCR技术克隆邻苯二酚-2,3-双加氧酶基因,生物信息学分析结果表明:C23O基因所编码的氨基酸序列具有双加氧酶基因保守结构域,编码的蛋白质也是一种稳定的酸性亲水蛋白质,分子量为35.01k D,等电点为5.45,二级结构由17%α-螺旋、36%β-折叠和45%的无规则卷曲组成,具有跨膜结构,不具有信号肽.三级结构预测结果与二级结构一致,三级结构有Fe2+结合位点.
[Abstract]:Response surface method was used to design the experiment. The pH value, temperature and days were selected as the influencing factors. The degradation conditions of wp3-1 strain were optimized for 50 mg/L inorganic salt medium with pyrene concentration. The nonlinear regression equation between the degradation rate of pyrene and the three factors was obtained. The optimum technological conditions of pyrene degradation were determined as follows: at the temperature of 34.15? When CP H value was 6.98 and degradation time was 4.28 days, the predicted degradation rate of pyrene was 86.42and the experimental verification value was 85.70. The results showed that the established model had high accuracy. The catechol -2o 3-dioxygenase gene was cloned by PCR technique. The results of bioinformatics analysis showed that the amino acid sequence encoded by the 1: C23O gene had a conserved domain of the double oxygenase gene, and the encoded protein was also a stable acidic hydrophilic protein. The molecular weight is 35.01kD, the isoelectric point is 5.45.The secondary structure is composed of 17% 伪 -helix 36% 尾 -fold and 45% irregular crimp. It has a transmembrane structure and no signal peptide. The predicted results of the tertiary structure are consistent with the secondary structure, and the tertiary structure has Fe2 binding site.
【作者单位】：新疆大学生命科学与技术学院;
【基金】：国家自然科学基金(31460027)
【分类号】：X172

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