Optimization for degradation of total petroleum hydrocarbon by the strain L-2 with response surface methodology

doi:10.16085/j.issn.1000-6613.2019-0520

Abstract

Abstract:

The degradation of total petroleum hydrocarbon(TPH) was studied by using L-2 strains isolated from oil-contaminated soil. L-2 was identified as Enterobacter sp. by 16S rRNA. Response surface method(RSM) based on box-behnken design(BBD) was adopted to evaluate the interaction effects of temperature, pH and TPH concentration on TPH degradation. A quadratic regression model was established to determine the best combination of the three experimental factors and the validity of the model was verified by analysis of variance(ANOVA). The value of determination coefficient (R²=0.9943) indicated a satisfactory agreement between the quadratic model and the experimental data. In the temperature of 30℃, pH of 7.13 and TPH concentration of 3.93g/L, the best degradation rate was 87.79%. It was found that TPH degradation rate was more significantly affected(p<0.0001) by temperature compared with other two parameters.GC/MS results indicated that TPH components in the range of hexadecane(C₁₆) to pentadecane (C₂₅) can be degraded ty strain L-2.

Key words: Enterobacter sp., bioremediation, total petroleum hydrocarbon(TPH), optimization, GC/MS

摘要：

从石油烃污染土壤中分离得到菌株L-2，对石油烃（TPH）的降解进行研究。通过16S rRNA鉴定L-2为肠杆菌属。采用基于Box-Behnken设计（BBD）的响应面方法（RSM），考察温度、pH和TPH质量浓度3个自变量对TPH降解率的交互效应，建立二次回归方程预测模型，确定3个实验因素的最佳组合，通过方差分析验证该二次模型的有效性。确定系数R²=0.9943，表明二次模型与实验数据吻合较好。响应面优化得到最佳降解条件：温度为30℃，pH为7.13和TPH质量浓度为3.93g/L，TPH降解率为87.79%。结果表明，温度对TPH降解速率的影响（P<0.0001）较其他两个参数更显著。GC/MS结果表明菌株L-2可降解十六烷(C₁₆)至二十五烷(C₂₅)范围内的TPH组分。

关键词: 肠杆菌属, 生物修复, 石油烃, 优化, 气相色谱/质谱

CLC Number:

Shuo SUN,Qiyou LIU,Shuiquan CHEN,Chaocheng ZHAO,Wenhe YU. Optimization for degradation of total petroleum hydrocarbon by the strain L-2 with response surface methodology[J]. Chemical Industry and Engineering Progress, 2019, 38(12): 5512-5518.

孙烁,刘其友,陈水泉,赵朝成,于文赫. 利用响应面法对L-2菌株降解石油烃进行优化[J]. 化工进展, 2019, 38(12): 5512-5518.

Figures/Tables 13

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实验编号	温度/℃	pH	TPH质量浓度 /g·L^-1	降解率/%
实验编号	温度/℃	pH	TPH质量浓度 /g·L^-1	实际值	预测值
1	30	7	2	74.85	74.67
2	35	7	4	69.37	69.62
3	35	6	6	25.68	27.28
4	35	6	2	33.26	31.65
5	35	7	4	68.98	69.62
6	35	8	2	28.56	31.65
7	40	7	6	18.26	20.23
8	35	7	4	64.56	69.62
9	35	7	4	73.89	69.62
10	30	8	4	67.88	69.27
11	35	7	4	71.3	69.62
12	40	6	4	20.58	19.19
13	35	8	6	30.36	26.32
14	30	7	6	70.8	70.31
15	40	8	4	4.09	4.81
16	40	7	2	25.89	24.59
17	30	6	4	55.6	54.88

实验编号	温度/℃	pH	TPH质量浓度 /g·L^-1	降解率/%
实验编号	温度/℃	pH	TPH质量浓度 /g·L^-1	实际值	预测值
1	30	7	2	74.85	74.67
2	35	7	4	69.37	69.62
3	35	6	6	25.68	27.28
4	35	6	2	33.26	31.65
5	35	7	4	68.98	69.62
6	35	8	2	28.56	31.65
7	40	7	6	18.26	20.23
8	35	7	4	64.56	69.62
9	35	7	4	73.89	69.62
10	30	8	4	67.88	69.27
11	35	7	4	71.3	69.62
12	40	6	4	20.58	19.19
13	35	8	6	30.36	26.32
14	30	7	6	70.8	70.31
15	40	8	4	4.09	4.81
16	40	7	2	25.89	24.59
17	30	6	4	55.6	54.88

类型	平方和SS	自由度DF	均方值	F	P
模型	8312.85	9	1052.51	134.19	＜0.0001
A	1762.99	1	5015.51	643.23	＜0.0001
B	3.62	1	2.24	0.29	0.6088
C	1013.40	1	38.11	4.99	0.0627
AB	43.56	1	206.93	26.54	0.0013
AC	485.76	1	3.20	0.41	0.5419
BC	116.42	1	22.00	2.82	0.1369
A2	79.27	1	224.30	28.77	0.0010
B2	1196.03	1	2691.65	345.20	＜0.0001
C2	3272.58	1	931.18	119.42	＜0.0001
残差	378.21	7	7.80	—	—
失拟项	162.26	3	2.48	0.21	0.8842
纯误差	215.94	4	11.78	—	—
总和	8691.06	16	—	—	—

类型	平方和SS	自由度DF	均方值	F	P
模型	8312.85	9	1052.51	134.19	＜0.0001
A	1762.99	1	5015.51	643.23	＜0.0001
B	3.62	1	2.24	0.29	0.6088
C	1013.40	1	38.11	4.99	0.0627
AB	43.56	1	206.93	26.54	0.0013
AC	485.76	1	3.20	0.41	0.5419
BC	116.42	1	22.00	2.82	0.1369
A2	79.27	1	224.30	28.77	0.0010
B2	1196.03	1	2691.65	345.20	＜0.0001
C2	3272.58	1	931.18	119.42	＜0.0001
残差	378.21	7	7.80	—	—
失拟项	162.26	3	2.48	0.21	0.8842
纯误差	215.94	4	11.78	—	—
总和	8691.06	16	—	—	—

统计项目	数值
标准偏差	2.86
平均值	47.29
偏差系数CV/%	6.06
预测误差平方和	193.58
确定系数R²	0.9943
调整系数R²	0.9862
预测系数R²	0.9797
精密度	38.017