淡水真菌Acremonium tubakii BMC-58产多肽cephaibol A的发酵工艺及抗菌活性  

Fermentation process and antibacterial activity of peptide cephaibol A produced by freshwater-derived fungus Acremonium tubakii BMC-58

作　　者：王路生 王欣茹 许芙蓉 范思聪 宗世坤 吴成柱 朱美林 WANG Lusheng;WANG Xinru;XU Furong;FAN Sicong;ZONG Shikun;WU Chengzhu;ZHU Meilin(Teaching and Research Section of Medicinal Chemistry,School of Pharmacy,Bengbu Medical College,Bengbu 233030,China;Anhui Biochemical Drug Engineering Technology Research Center)

机构地区：[1]蚌埠医学院药学院药物化学教研室,蚌埠233030 [2]安徽省生化药物工程技术研究中心

出　　处：《山西医科大学学报》2023年第7期988-995,共8页Journal of Shanxi Medical University

基　　金：国家自然科学基金青年基金资助项目(81903486);安徽省自然科学基金青年基金资助项目(1908085QH347);蚌埠医学院研究生创新项目(Byycx21033)。

摘　　要：目的 改良淡水真菌Acremonium tubakii BMC-58产多肽cephaibol A的发酵工艺,提高其产率并检测其抗菌活性。方法 通过对6种培养基的筛选确定基础培养基后,采用单因素实验研究接种量、培养温度、摇床转速、培养时间、初始pH、碳源、氮源和无机盐对菌株BMC-58产cephaibol A产率的影响,确定最适的培养条件;接下来采用Plackett-Burman实验、最陡爬坡实验和响应面试验优化培养基成分;最后通过纸片法检测cephaibol A对4种细菌(Methicillin-resistant Staphylococcus aureus,MRSA;Staphylococcus aureus,S.aureus;Bacillus subtilis,B.subtilis;Mycobacterium phlei,M.phlei)的抗菌活性。结果 确定了菌株BMC-58在真菌2号培养基中产多肽cephaibol A的产率最高,并将培养基成分调整为蛋白胨、酵母浸膏、玉米淀粉、可溶性淀粉、MgSO_(4)、CaCO_(3)、谷氨酸钠和甘露醇,培养条件为菌株接种量2%,培养基初始pH6.0,培养温度28℃,摇床转速180 r/min以及培养时间7 d,最终cephaibol A的产率(17.23 mg/L)比优化前(8.7 mg/L)提高了1.97倍。同时抗菌实验中cephaibol A对MRSA抗菌活性较好,抑菌圈为9.0 mm。结论 通过优化培养条件后,cephaibol A的产率具有一定的提高,对检测的4种细菌也都有较好的抗菌活性,是一种潜在的抗耐药菌药物。Objective To increase the production of the polypeptide cephalibol A produced by freshwater fungal strain Acremonium tubakii BMC-58 by improving the fermentation process,and investigate the antibacterial activity of cephaibol A.Methods The basal culture medium was screened from the six media,and then the culture conditions including inoculation amount,culture temperature,rotation speed,culture time,initial pH,carbon source,nitrogen source,and inorganic salts were screened by single factor test.The optimum fermentation medium components were determined by the Plackett-Burman test,the steepest climbing experiment,and the response surface analysis method of Box-Behnken design.The antibacterial activity of cephalibol A against four types of bacteria(Methicillin-resistant Staphylococcus aureus,MRSA;Staphylococcus aureus,S.aureus;Bacillus subtilis,B.subtilis;Mycobacterium phlei,M.phlei)was tested by the paper disk method.Results The medium No.2 was chosen to be the basic medium,and the yield of cephalibol A was the highest.The optimum culture medium for Acremonium tubakii BMC-58 contained peptone,yeast extract,corn starch,soluble starch,MgSO_(4),CaCO_(3),sodium glutamate,and mannitol.The optimal fermentation conditions were 2.0%(V/V)of inoculum amount,initial pH 6.0,28℃of culture temperature,180 r/min of rotation speed,and 7 d of culture time.After optimization,the yield of cephalibol A was increased from 8.7 mg/L to 17.23 mg/L,which was 1.97 times that before optimization.The antibacterial test showed that cephalibol A exhibited the strongest activity against MRSA with a 9.0 mm antibacterial zone.Conclusion After optimization,the yield of cephalibol A is improved.The cephalibol A has a better antibacterial activity against four bacteria,and it can be a potential drug against drug-resistant bacteria.

关 键 词：淡水真菌 cephaibol A 发酵工艺 抗菌肽 培养条件 

分 类 号：R914[医药卫生—药物化学]