ARTP-UV复合诱变结合含盐平板选育产耐盐高活性蛋白酶的米曲霉

Combined Mutagenesis by Atmospheric and Room Temperature Plasma and Ultraviolet and Screening on Salt-Containing Plates of Aspergillus oryzae for High Salt Tolerance and Activity of Protease

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摘要对从高盐稀态酱醪中筛选得到的米曲霉(Aspergillus oryzae)K9进行常压室温等离子体(atmospheric and room temperature plasma,ARTP)复合紫外诱变,并利用高盐含量平板进行驯化培养和筛选,旨在得到产耐盐高活性蛋白酶的菌株。首先对K9进行ARTP诱变,筛选得到优势菌株S-G27,其中性蛋白酶活性较K9提高35.82%;接着以S-G27为出发菌株进行紫外诱变,筛选得到优势菌株G-U2,其中性蛋白酶活性较K9提高56.81%;G-U2所产蛋白酶的耐盐性进一步提高,在15%NaCl溶液中,G-U2中性蛋白酶活性残留率为K9的1.94倍;G-U2稳定性良好,第7代时菌株的中性蛋白酶活性残留率为92%;和出发菌株K9相比,G-U2的菌丝更短、孢子更多、孢子体积更大且表面纹饰更多;诱变未引起黄曲霉毒素相关基因的表达,符合生产安全要求。在模拟发酵酱油实验中,发酵终点G-U2中性蛋白酶活性、酸性蛋白酶活性、氨基酸态氮质量浓度分别较K9高出120.19%、52.29%、18.37%。 This study aimed to obtain strains producing high levels of salt-tolerant protease from Aspergillus oryzae K9,isolated from high-salt liquid-state moromi,by combined mutagenesis induced by atmospheric and room temperature plasma(ARTP)and ultraviolet(UV)followed by domestication and screening using high-salt plates.The dominant mutant S-G27,whose neutral protease activity was increased by 35.82%compared with K9,was obtained after ARTP treatment of K9.The dominant mutant G-U2,whose neutral protease activity was increased by 56.81%compared with K9,was obtained after subsequent UV treatment of S-G27.In 15%NaCl solution,the residual activity of neutral protease from G-U2 was 1.94 times as high as that of K9.G-U2 exhibited good stability,and 92%of the initial enzyme activity remained after the seventh passage.Compared with the starting strain K9,G-U2 showed shorter mycelia,more spores,larger spore volume,and more surface ornaments.Mutagenesis did not induce the expression of aflatoxin-related genes,meeting production safety requirements.In simulated soy sauce fermentation experiments,the neutral protease activity,acid protease activity,and amino nitrogen content of G-U2 at the end of fermentation were 120.19%,52.29%,and 18.37%higher than those of K9,respectively.

作者孙思城李浩然苏国万吴惠贞林伟锋 SUN Sicheng;LI Haoran;SU Guowan;WU Huizhen;LIN Weifeng(College of Food Science and Engineering,South China University of Technology,Guangzhou 510640,China;Heshan Donggu Flavoring Food Co.Ltd.,Jiangmen 529738,China)

机构地区华南理工大学食品科学与工程学院鹤山市东古调味食品有限公司

出处《食品科学》北大核心 2025年第15期120-127,共8页 Food Science

基金国家自然科学基金面上项目(32172329)。

关键词米曲霉常压室温等离子体诱变紫外线诱变耐盐蛋白酶 Aspergillus oryzae atmospheric and room temperature plasma mutagenesis ultraviolet mutagenesis salt tolerant protease

分类号 TS264.2 [轻工技术与工程—发酵工程]