基于响应面法的耐镉假单胞菌TCd-1培养条件优化  

Optimization of culture conditions of Cd-tolerant strain Pseudomonas TCd-1 based on response surface methodology

作　　者：汪敦飞 朱胜男 肖清铁 郑新宇[1,2] 林瑞余 WANG Dunfei;ZHU Shengnan;XIAO Qingtie;ZHENG Xinyu;LIN Ruiyu(Fujian Provincial Key Laboratory of Agroecological Processing and Safety Monitoring,College of Life Sciences,Fujian Agriculture and Forestry University,Fuzhou 350002,Fujian,China;Key Laboratory of Crop Ecology and Molecular Physiology of Fujian Province University,Fujian Agriculture and Forestry University,Fuzhou 350002,Fujian,China)

机构地区：[1]福建农林大学生命科学学院,福建省农业生态过程与安全监控重点实验室,福建福州350002 [2]福建农林大学,作物生态与分子生理学福建省高校重点实验室,福建福州350002

出　　处：《浙江农林大学学报》2020年第5期914-921,共8页Journal of Zhejiang A&F University

基　　金：“十三五”国家重点研发计划项目(2017YFD0800900);科技部科技基础性工作专项(2015FY111300)。

摘　　要：【目的】利用响应面法优化耐镉菌株假单胞菌Pseudomonas TCd-1的培养条件。【方法】以基础发酵培养条件为对照,菌株吸光度D(660)作为评价指标,利用单因素试验确定培养基组分的最佳碳源、氮源和无机盐种类,通过Plackett-Burman试验设计评价牛肉膏、酵母粉、氯化镁、培养温度、初始pH、接菌量、培养时间和转速8个因素对菌株生长的影响,进而确定显著影响因素,然后进行最陡爬坡试验获得显著影响因素的最优组合,结合Box-Behnken试验设计和响应面分析,优化假单胞菌TCd-1的培养条件。【结果】菌株培养基组分的最佳碳源、氮源和无机盐分别为牛肉膏、酵母粉和氯化镁;影响菌株生长的显著性因素为酵母粉、培养温度、初始pH;基于响应面法优化后的培养条件为:牛肉膏质量分数为0.5%,酵母粉1.0%,氯化镁0.5%,pH 6.3,温度33℃,接菌量1.25%,转速160 r·min^−1,培养时间24 h。【结论】根据最佳条件,进行重复试验得到优化后的菌液吸光度D(660)比对照提高了67.07%,与模型预测值相一致,表明优化后的条件显著促进了菌株的生长,达到预期目的。[Objective]This study aims to optimize the culture conditions of Cd-tolerant Pseudomonas TCd-1 based on response surface methodology.[Method]Taking the basic fermentation conditions as the control,and the absorbance D(660)value of the strain as the evaluation index,the single factor test was carried out to determine the best carbon source,nitrogen source,and inorganic salt species of the culture medium components.The Plackett-Burman test design was used to evaluate the influence of 8 factors on the growth of the strain,including beef extract,yeast powder,magnesium chloride,culture temperature,initial pH,inoculation quantity,culture time,and rotation speed.The best combination of significant factors was obtained by the steepest climbing test.The culture conditions of TCd-1 were optimized by combining the box Behnken design test and response surface analysis.[Result]The best carbon source,nitrogen source,and inorganic salt of the strain culture medium components were beef extract,yeast powder,and magnesium chloride.The significant factors affecting the growth of the strain were yeast powder,culture temperature,and initial pH.The optimized culture conditions based on response surface methodology included beef extract 0.5%,yeast powder 1.0%,magnesium chloride 0.5%,pH 6.3,temperature 33℃,inoculation rate 1.25%,rotation speed 160 r·min−1,and culture time 24 h.[Conclusion]According to the optimal conditions,the D(660)value of the optimized bacterial solution is 67.07%higher than that of the control after repeated experiments,which is consistent with the predicted value of the model,indicating that the optimized conditions could significantly promote the growth of the strain and achieve the desired goal.

关 键 词：假单胞菌 镉 PLACKETT-BURMAN设计 优化培养 响应面法 

分 类 号：Q935[生物学—微生物学]