平菇高产漆酶和锰过氧化物酶液体培养条件的优化  

Optimization of Liquid Culture Conditions for High Yield Laccase and Manganese Peroxidase in Pleurotus Ostreatus

作　　者：黄娇 孙冬梅 柯丽霞 HUANG Jiao;SUN Dong-mei;KE Li-xia(College of Life Sciences,Anhui Normal University,Anhui Provincial Key Laboratory of the Conservation and Exploitation of Biological Resources,Wuhu 241000,China)

机构地区：[1]安徽师范大学生命科学学院重要生物资源保护与利用研究安徽省省级重点实验室

出　　处：《安徽师范大学学报（自然科学版）》2019年第6期555-561,共7页Journal of Anhui Normal University(Natural Science)

基　　金：安徽省自然科学基金项目(1808085MC55)

摘　　要：在以油菜秸秆为主要基质的液体培养基中,研究了不同金属离子(Cu^2+、Mn^2+、Al^3+、Zn^2+和Fe^3+,浓度均为0.5mmol/L)、不同浓度有机酸盐及吐温80对平菇分泌木质素降解酶系活性的影响。结果表明:发酵第8天,酶活达到峰值。Cu^2+、Mn^2+、Al^3+都能显著提高平菇分泌的漆酶和锰过氧化物酶的活性;Zn^2+对漆酶活性则有显著抑制作用。1mmol/L草酸钠和1mmol/L、3mmol/L、5mmol/L甘醇酸钠对漆酶和锰过氧化物酶的活性有显著的促进作用。0.05%、0.1%和0.15%的吐温80可显著提高漆酶的活性,0.1%、0.15%和0.20%的吐温80可显著提高锰过氧化物酶的活性。以不同金属离子、不同浓度甘醇酸钠、不同浓度吐温80、不同酵母菌为实验因素进行正交实验。正交实验结果表明,利用平菇液体发酵油菜秸秆获得较高漆酶活性的适宜培养条件为:在平菇与产朊假丝酵母混菌液态发酵体系中,加入0.5mmol/LCu^2+、3mmol/L甘醇酸钠及0.05%吐温80,漆酶的活性最高为851.4U/L。平菇发酵油菜秸秆获得较高锰过氧化物酶活性的适宜培养条件为:平菇与产朊假丝酵母混菌液态发酵体系中,加入0.5mmol/L Mn^2+、5mmol/L甘醇酸钠及0.1%吐温80;而最优培养条件下,锰过氧化物酶活性为478.3U/L。本研究探索出一种平菇高产木质素降解酶系的低成本培养方法。The effects of different metal ions(Cu^2+,Mn^2+,Al^3+,Zn^2+and Fe^3+,their concentration were all 0.5 mmol/L),Organic acid salt and Tween 80 in vitro production of extracellular laccase(Lac)and manganese peroxidase(MnP)activities by Pleurotus ostreatus,which was liquid-cultivated in a medium with rape straw as a primary nutrient were investigated.Lac and MnP activities both reached the peak values in the 8 th day.Lac and MnP activities were both significantly elevated by metal ions such as Cu^2+,Mn^2+,and Al^3+;but Zn^2+significantly decreased Lac activities.1 mmol/L sodium oxalate and 1 mmol/L,3 mmol/L,5 mmol/L sodium glycollate significantly increased the activities of MnP and Lac.0.05%,0.1%and 0.15%Tween 80 could significantly increase Lac activities;0.1%,0.15%and 0.2%Tween 80 could significantly increase MnP activities.The kinds of metal ions,different concentration of sodium glycollate,different concentration of Tween 80 and the kinds of yeasts were used as the design elements of orthogonal experiment.Orthogonal experiment results showed that the optimal culture condition for Lac production was:0.5 mmol/L Cu^2+,3 mmol/L sodium glycollate,0.05%Tween 80 were added into the mixed-strain fermentation system of P.ostreatus and utilis in liquid-state rape straw medium;the activity of Lac was 851.4 U/L under this culture condition.The optimal culture condition for MnP production was:0.5 mmol/L Mn^2+,5 mmol/L sodium glycollate,0.1%Tween 80 were added into the mixed-strain fermentation system of P.ostreatus and Candida utilis in liquid-state rape straw medium;the MnP activities was 478.3 U/L under this culture condition.This study explores a low-cost culture method for high-yield lignin-degrading enzymes of P.ostreatus.

关 键 词：平菇 外源因素 漆酶 锰过氧化物酶 培养条件优化 

分 类 号：Q935[生物学—微生物学]