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

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CGMCC 3.316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23 IU/mL, 与出发菌株CGMCC 3.316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。

Abstract:

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 beta-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.