黑曲霉利用五倍子固体发酵产单宁酶的条件研究.doc

黑曲霉 B0201 利用五倍子固体发酵产单宁酶的条件研究 摘 要: 通过比较常规灭菌发酵和生料发酵, 研究黑曲霉 B0201 利用五倍子固体发酵产单宁酶的条件。结果表明, 在生料发酵过程中, 采用20%五倍子并且以(NH4)2SO4为氮源制备单宁酶的最佳条件为: 液固比=1.6:1、温度 30°C、初始 pH 6.0。在该条件下通过 96 h 的培养, 单宁酶的活力可达 51.2 U/gds, 是常规灭菌发酵的 3.6 倍。以上结果显示, 生料发酵生产单宁酶是一种高效可行的方法。 关键词: 黑曲霉, 单宁酶, 生料发酵, 五倍子, 固体发酵 Research on Conditions of Tannase Production from Aspergillus Niger B0201 Under Solid-state Fermentation Using Gallnut LI Yang-Zhen 1,2 QIU Shu-Yi 1,2* BAO Yu-Xin 1,3 Wu Xin-Ying 1,2 (1. Guizhou Province Key Laboratory of Fermentation Engineering and Biopharmacy, Guiyang, Guizhou 550003, China) (2. Chemical Engineering College of Guizhou University, Guiyang, Guizhou 550003, China) (3. Life Science College of Guizhou University, Guiyang, Guizhou 550000, China) Abstract: According to the comparative research of conventional sterile fermentation and uncooked material fermentation, the conditions of tannase production from Aspergillus Niger B0201 under solid-state fermenta- tion using gallnut was studied. The results showed that the liquid?solid ratio of 1.6: 1, temperature of 30°C, initial pH of 6.0 were the optimal conditions for tannase production under uncooked material fermentation with gallnut content of 20% and (NH 4 ) 2 SO 4 for nitrogen sources. Under this condition, tannase activity reached to 51.2 U/gds after 96 h of cultivation, which was 3.6 times as the conventional sterile fermentation. The results above revealed that under uncooked material fermentation for tannase production was a feasible method with high efficiency. Keywords: Aspergillus niger, Tannase, Uncooked material fermentation, Gallnut, Solid-state fermentation1124 微生物学通报 2009, Vol.36, No.8 /wswxtbcn 单宁酶, 可水解没食子单宁中的酯键和缩酚羧 键, 生成没食子酸和葡萄糖 [1] 。单宁酶可应用于制 革、酿酒、医药、饮料等领域, 具有广阔的应用前 景。但是目前生产单宁酶的效率不高, 市场价格昂 贵, 阻碍了单宁酶的应用。单宁酶是一种诱导酶, 主 要利用微生物发酵生产。目前国内主要采用液体发 酵法生产单宁酶 [2,3] , 而国外近期的研究主要集中在 固体发酵法 [4?6] 。很多资料 [7? 显示固体发酵主要产 胞外单宁酶, 容易提取, 稳定性好, 杂酶少, 且设备 比较简单, 同时固体发酵有利于提高产酶活力。 本实验室筛选出的黑曲霉 B0201 菌株能利用中 国特产五倍子作为诱导物固体发酵产单宁酶。但是 在试验过程中发现原料五倍子在高压蒸汽灭菌时很