煤附生真菌产漆酶菌株的分离鉴定及产酶特性研究kdh.pdf

Mycosystema菌物学报15May201029(3):[e-mail]1672-6472CN11-5180Q©2010InstituteofMicrobiologyCASallrightsreserved.煤附生真菌产漆酶菌株的分离鉴定及产酶特性研究陈今朝1王剑锋2*李江2饶军2秦家顺11长江师范学院生物系涪陵4081002东华理工大学生物系抚州344000摘要：从煤炭样品中筛选到一株产漆酶活性菌株经菌体形态观察和ITS序列分析鉴定为TrichodermaasperellumW03。菌株所产漆酶的最适反应pH为3.5－4.5最适反应温度45℃类似于白腐真菌漆酶。液态发酵条件的均匀设计实验表明适宜的发酵培养基组成为：土豆200.00g/L、葡萄糖9.36g/L、米糠粉37.44g/L、硝酸钾4.00g/L、KH2PO43.20g/L、MgSO4·7H2O2.00g/L、CuSO4·5H2O0.005g/L、初始pH8.0；在33℃、180r/min、50mL/250mL的摇瓶培养条件下棘孢木霉W03在孢子接种培养后48h、84h产酶量较高分别处在菌体的快速生长期和衰亡期；菌体产酶受Cu2+、联苯胺诱导而受1-萘酚、愈创木酚和24-D抑制。关键词：均匀设计半知菌棘孢木霉ITS序列Identificationandproductionoflaccase-producingfungalstrainisolatedfromcoalCHENJin-Zhao1WANGJian-Feng2*LIJiang2RAOJun2QINJia-Shun11DepartmentofBiologyYangtzeNormalUniversityFuling408100China2DepartmentofBiologyEastChinaInstituteofTechnologyFuzhou344000ChinaAbstract:Laccase-producingstrainscreenedfromlow-rankcoalwasidentifiedasTrichodermaasperellumW03accordingtoitsmyceliamorphologyandITSsequenceanalysis.ThelaccasehasanoptimumpHof3.5－4.5andanoptimumtemperatureof45℃whichissimilartothatfromwhiterotfungi.Componentsoflaccase-producingmedium(g/L)optimizedbyuniformdesignwere200.00g/Lpotato9.36g/Lglucose37.44g/Lricebranpowder4.00g/LKNO33.20g/LKH2PO42.00g/LMgSO4·7H2Oand0.005g/LCuSO4·5H2O.Thepeakoflaccaseproductionappearedinthe48hand84hcorrespondingtothelogarithmicphaseanddeclinephaserespectivelywhenTrichodermaasperellumW03wassubmergedinshaking-flaskwithliquidvolume50mL/250mLunderconstanttemperatureof33℃andvibrationof180r/min.Thelaccaseproduction