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海洋芽孢杆菌N11-8产蛋白酶的发酵条件优化 被引量:4

Optimization of Fermentation Conditions of Bacillus sp. N11-8 on the Production of Protease PBN11-8
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摘要 本研究采用响应面法(RSM)对南极海洋芽孢杆菌(Bacillussp.)N11-8液体发酵产蛋白酶PBN11-8的发酵条件进行快速优化。通过单因素实验初步确定南极海洋芽孢杆菌N11-8产蛋白酶的最佳碳源和氮源分别为可溶性淀粉和蛋白胨;并通过Plackett-Burman(PB)设计对影响其产酶相关因素进行评估,筛选出具有显著效应的3个因素:温度、氮源和碳源;以最陡爬坡实验逼近至上述因子最大响应区域,进而采用RSM法对其最佳水平范围进行研究,确定最优发酵条件为:可溶性淀粉3 g/L,蛋白胨13.1 g/L,酵母浸粉2.9 g/L,Na Cl_5 g/L,KH_2PO_4 1 g/L,Fe Cl3·6H_2O_2 mmol/L,初始pH值为7.0,温度为34.0℃,转速为200 r/min,装液量为50 ml/250 ml,接种量为4%,培养时间为60 h。最终优化后的酶活达到90.5 U/ml,比初始酶活提高了23.6%。 In this study,we optimized fermentation conditions to increase the production of PBN11-8 by Bacillus sp.N11-8.Response surface methodology(RSM)is a generally available method used to optimize the fermentation conditions of proteases,by fitting factors function relation between response value.The optimal carbon and nitrogen sources for protease production by Bacillus sp.N11-8were starch and peptone respectively,which were verified in single-factor experiments.Other optimal fermentation conditions were determined as follows:temperature35~C,incubation time60h,rotational speed 200r/min,pH7,initial inoculums4%,culture volume50ml in a250ml shake flask,and starch,peptone, yeast extract,NaC1,KH2PO4,and metal ions(Fe3+)concentrations of3g/L,15g/L,2.5g/L,5.0g/L, 1.0g/L,and2.0mmol/L,respectively.The Plackett-Burman design(PB)method was used to evaluate factors affecting protease production by Bacillus sp.N11-8,and three significant factors were identified: temperature,peptone,and yeast extract.Next,we used the steepest ascent design to approach the maximum response area of the three factors.Furthermore,the best scope of fermentation conditions was studied using a central composite test design,which is a type of RSM.Finally,through analysis of variance for the RSM,we obtained a regression equation and calculated the theoretical optimal fermentation conditions as follows:3g/L of soluble starch,13.1g/L of peptone,2.9g/L of yeast extract, 5g/L ofNaC1,1g/L of KI-I2PO4,2mmol/L of Fe3~,pH7.0,temperature34℃,rotational speed200r/rain, culture volume50ml/250ml,initial inoculums of4%,incubation time60h.Under the optimized fermentation conditions,the theoretical enzyme activity reached90.8U/ml,and in the verification test, enzyme activity reached90.5U/ml,which increased23.6%of the initial enzyme activity.
作者 朱祥杰 王震 苑志欣 郝建华 郑兰红 ZHU Xiangjiel;WANG Zhen;YUAN Zhixin;HAO Jianhua;ZHENG Lanhong(Key Laboratory of Sustainable Development of Polar Fishery, Ministry of Agriculture and Rural Affairs, Key Laboratory of Sustainable Development of Marine Fisheries, Ministry of Agriculture and Rural Affairs, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071;College of Fisheries and Life Science,Shanghai Ocean University,Shanghai201306)
出处 《渔业科学进展》 CSCD 北大核心 2018年第6期155-163,共9页 Progress in Fishery Sciences
基金 中国水产科学研究院基本科研业务费(2014801YQ01) 山东省科技发展计划项目(2014GsFl21016)共同资助.
关键词 芽孢杆菌 蛋白酶 PLACKETT-BURMAN设计 响应面法 Bacillus sp. Protease Plackett-Burman design RSM
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