富产琥珀酸米曲霉菌株的诱变选育及初步应用-生物工程专业论文.docxVIP

I I 摘 要 琥珀酸作为酱油的重要呈味物质，可以增加酱油的鲜味，对提高酱油的品质有 着重要意义。米曲霉 AS3.042 作为我国酱油生产普遍使用的菌株，发酵过程中产生 微量的琥珀酸，成品中的琥珀酸浓度很低，提高琥珀酸在酱油中的含量对酱油品质 的提升非常重要。 因此本文以米曲霉 AS3.042 为出发菌株，经紫外线和亚硝基胍诱变后，筛选出 一支富产琥珀酸菌株 ASC-5，相同条件下，摇瓶培养发酵液中琥珀酸含量为 3.60 g/L， 比未诱变原始出发菌株 AS3.042 提高了 370 倍，琥珀酸的遗传性状稳定。 通过对 ASC-5 菌落特征和菌株形态特征的观察，初步鉴定该菌株为半知菌亚门 （Deuteromycotina）、丝孢纲（Hyphomycetes）、丝孢目(Hyphomycetes)、从梗孢科 （Moniliaceae）、曲霉属(Aspergillus)。 然后通过对菌株 ASC-5 固体制曲，得出其固体曲中琥珀酸质量为 19.20 g/Kg， 相同条件下琥珀酸含量比诱变前菌株 AS3.042 有明显提高；酸性蛋白酶、淀粉酶、 纤维素酶活分别为 93.10 u/g、220.73 u/g、8.93 u/g，比诱变前菌株 AS3.042 固体曲中 酶活提高了 21.7 %、20.4 %、4.1%；中性蛋白酶、半纤维素酶活分别为 73.40 u/g、 7.34 u/g，比诱变前菌株 AS3.042 下降了 7.0 %、6.9 %。 最后按照完全相同的制曲工艺，分别制备米曲霉AS3.042和ASC-5固体曲。然后 将上述两种曲分别在容积为5 L的酱油酿造池中，相同的工艺条件下发酵，酿造结束 后制备得到不同样品酱油，分析评价样品酱油感官特征和理化指标。结果表明：以 ASC-5为菌株酿制的样品酱油感官特征明显优于米曲霉AS3.042，琥珀酸含量为0.97 g/L，达到其口味阈值，且其他理化指标均符合GB18186-2000规定且优于米AS3.042。 为ASC-5进一步放大研究奠定基础。 关键词：米曲霉；诱变；琥珀酸产生菌；酱油酿造 II II Abstract As an important flavoring material of soy sauce, succinic acid plays a significant role in the increase of flavor and the improvement of quality of soy sauce. Aspergillus oryzae AS3.042 is a widely used strain in the production process of soy sauce in our country, however, only traces of Succinic acid is produced in the fermentation process of soy sauce. As a result, the concentration of succinic acid in the finished product is very low. Thus, it appears so important to increase the content of succinic acid in order to improve the quality of soy sauce. Considering the needs of above results, Aspergillus oryzae AS3.042 was used as a starting strain in this paper, and a strain named ASC-5 which has a relatively high yield of succinic acid was bred by the method of UV mutagenic and nitrosoguanidine mutagenic. The succinic acid yield fermented by the strain ASC-5 reached 3.60 g/L in the same fermentation conditions, which was 370-folds higher compared with the yield of the starting strain AS3.042, meanwhile, the inheritable character of succinic acid is stable. Through the observati