响应面法优化芘降解菌的降解条件及降解基因分析

晁群芳; 赵亚光; 吴盼云; 徐苗; 段魏魏; 肖路梅; 马腾飞

doi:10.13568/j.cnki.651094.2017.02.002

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响应面法优化芘降解菌的降解条件及降解基因分析

更新时间：2026-01-21

- 响应面法优化芘降解菌的降解条件及降解基因分析
- Journal of Xinjiang University (Natural Science Edition in Chinese and English) Vol. 34, Issue 2, Pages: 133-139(2017)
- 作者机构：
  
  新疆大学生命科学与技术学院
- 作者简介：
- 基金信息：
- DOI：10.13568/j.cnki.651094.2017.02.002
  CLC： X172
- Published：2017
- 稿件说明：
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[1]晁群芳,赵亚光,吴盼云,等.响应面法优化芘降解菌的降解条件及降解基因分析[J].新疆大学学报(自然科学版),2017,34(02):133-139.
[1]晁群芳,赵亚光,吴盼云,等.响应面法优化芘降解菌的降解条件及降解基因分析[J].新疆大学学报(自然科学版),2017,34(02):133-139. DOI： 10.13568/j.cnki.651094.2017.02.002.

DOI：10.13568/j.cnki.651094.2017.02.002.

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 Methodology(RSM)was employed to optimize some factors such as the culture temperature

p H value

and culture time

to enhance the biodegradation process. The degradation conditions of wp3-1 strain were optimized by using pyrene concentration 50 mg/L inorganic medium

and the nonlinear regression equation between pyrene degradation rate and the 3 factors was obtained. The optimum process parameters for pyrene-degrading rates were determined as the following: the incubation temperature of 34.15?C

the p H value of 6.98 and the culture time of 4.28 days. Under these optimum conditions

the predicted pyrene-degrading rate was 86.42%

whereas the observed value was 85.70%. Experimental results showed that the proposed model had higher accuracy. Catechol-2

3-dioxygenase gene(C23O)was cloned by PCR. Bioinformatic analytical results of the amino acid sequence indicated that there was dioxygenase conserved domain in this sequence

and the molecular weight of the protein component was 35.01 k D. The p I was 5.45

and it was stable protein. The prediction of the secondary structure showed that there were 17%α-helix

36% β-fold and 45% random coils. The subcellular localization signal peptide and transmembrane prediction and the signal analysis showed that C23O is transmembrane proteins

but do not contain a signal peptide. Teritary structure of C23O was similar with its secondary structure. From this model

we can know that there are Fe2+binding sites of C23O.

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