黑曲霉原生质体诱变选育β-葡萄糖苷酶高产菌株  

Protoplast mutagenesis for improving β-glucosidase production of Aspergillus niger

作　　者：王春丽[1,2] 武改红[2] 陈畅[1] 陈树林[2] 

机构地区：[1]北京化工大学生命科学与技术学院,北京100029 [2]中国科学院微生物研究所,北京100101

出　　处：《生物工程学报》2009年第12期1921-1926,共6页Chinese Journal of Biotechnology

基　　金：国家重点基础研究发展计划(973计划)(No.2007CB714304);国家高技术研究发展计划(863计划)(No.2009AA03Z221)资助~~

摘　　要：本研究报道了以原生质体诱变技术选育高产β-葡萄糖苷酶的黑曲霉菌株,并研究了其发酵特性。以黑曲霉CGMCC3.316为出发菌株,通过紫外诱变得到突变株3-3M。然后以3-3M为供试菌株,研究了其原生质体制备与再生的条件。最后通过原生质体诱变,选育得到一株β-葡萄糖苷酶活力较高的突变株60B-3D。该菌株具有良好的遗传稳定性,酶活力平均达到23IU/mL,与出发菌株CGMCC3.316相比提高39%。此外,该菌株的木聚糖酶活力也有所增加。同时考察了黑曲霉60B-3D的发酵特性,并与3-3M和出发菌株进行比较,结果表明该菌株有较高的蛋白分泌能力。本研究为发酵生产β-葡萄糖苷酶提供了一株良好的供试菌株。The aims of this research were to isolate a Aspergillus niger strain with higher β-glucosidase activity.We utilized the β-glucosidase producing strain Aspergillus niger CGMCC 3.316 as the original strain to first obtain a mutant 3-3M through ultraviolet irradiation.Then we studied the conditions of protoplast release and regeneration for strain 3-3M.We treated the protoplasts of strain 3-3M via ultraviolet irradiation and obtained another isolated mutant 60B-3D.The strain 60B-3D showed much higher β- glucosidase production than the original strain and 3-3M strain. The β-glucosidase activity of strain 60B-3D was 23.4 IU/mL, with an improvement of 39% compared with the original strain, and 23% compared with strain 3-3M. We also studied the fermentation process of strain 60B-3D, and compared it with the original strain and strain 3-3M. We found the strain 60B-3D exhibited an improvement in xylanase production. The comparison results also showed that the strain 60B-3D secreted more protein. These results were beneficial for producing β-glucosidase through this productive mutant.

关 键 词：黑曲霉 原生质体诱变 Β-葡萄糖苷酶 选育 木聚糖酶 

分 类 号：Q933[生物学—微生物学]