酶法制备黑曲霉原生质体的条件  

The conditions of preparing Aspergillus niger protoplasts by enzymolysis

作　　者：张婷[1] 李佳[1] 侯英敏[1] 孙玉梅[1] 

机构地区：[1]大连工业大学生物工程学院,辽宁大连116034

出　　处：《大连工业大学学报》2015年第4期239-242,共4页Journal of Dalian Polytechnic University

基　　金：辽宁省教育厅创新团队项目(LT2011008)

摘　　要：通过原生质体诱变可提高微生物的诱变率。研究了黑曲霉原生质体的制备条件,包括菌龄、渗透压稳定剂、裂解酶组成、酶解时间及再生培养基中的金属离子,并在相差显微镜下观察了黑曲霉原生质体的形成过程。综合考虑原生质体形成和再生的情况,确定了原生质体制备的最适条件为菌丝培养4d,以0.6mol/L NaCl作为渗透压稳定剂,用0.5%纤维素酶和0.5%蜗牛酶酶解3h。结果表明,最适再生培养基为酵母浸粉0.2%、蛋白胨0.5%、NaCl 0.2%、淀粉1%、CaCl20.1%、MgSO4·5H2O0.1%。与其他金属离子相比,钙离子对原生质体的再生具有明显的促进作用。Mutation efficiency of microbe can be improved by mutation of protoplasts.The preparation conditions of Aspergillus niger protoplast including mycelium incubation time,osmotic stabilizer,enzymes composition,enzymatic reaction time and metal ions were investigated,and the formation procedure of A.niger protoplast was observed under the phase contrast microscope.The optimal conditions were determined that the mycelia incubated for 4dand treated with 0.5% snailase and0.5% cellulase for 3hin 0.6 mol/L NaCl osmotic stabilizer.The result showed that most suitable regeneration media contained yeast 0.2%,peptone 0.5%,NaCl 0.2%,starch 1%,CaCl20.1% and MgSO4·5H2O 0.1%.Calcium ions could improve the regeneration of A.niger protoplast more effectively than the others.

关 键 词：黑曲霉 原生质体制备 原生质体再生 

分 类 号：TS201.3[轻工技术与工程—食品科学]