摘要
将实验室保藏的异常威客汉逊酵母(Wickerhamomyces anomalus)JM1-9进行多轮次的紫外-硫酸二乙酯(Ultraviolet-diethyl sulfate,UV-DES)复合诱变,通过比较透明圈大小及酶活高低筛选出一株高产酯酶菌株UVDES3-2,其酶活为(56.78±0.04)U/mL。该突变菌株与原始菌株((33.31±0.02)U/mL)相比,酶活提高了70.45%。再通过遗传稳定性试验,使其酶活稳定在(51.51±0.016)~(51.54±0.024)U/mL,与原始菌株相比提升了54.63%~54.72%。通过单因素试验确定影响因素大小为时间>初始pH值>发酵温度>装液量>接种量>摇床转速。利用正交试验确定最优发酵条件为时间24 h、温度为28℃、初始pH值为7、接种量为5%、转速为140 r/min、装液量为100/250 mL。在最佳发酵条件下,突变菌株UVDES3-2的酯酶酶活为(53.73±0.14)U/mL,比原始菌株JM1-9的酯酶酶活提高了61.30%。
A series of ultraviolet-diethyl sulfate(UV-DES)complex mutagenic mutation rounds were conducted on Wickerhamomyces anomalus JM1-9 samples stored in the laboratory.A high-yielding esterase strain,UVDES3-2,was then screened by comparing the size of the transparent ring and the level of enzyme activity.The enzyme activity was found to be(56.78±0.04)U/mL.In comparison to the original strain,which exhibited an enzyme activity of(33.31±0.02)U/mL,the mutant strain demonstrated a 70.45%increase in enzyme activity.A genetic stability test demonstrated that the enzyme activity of the strain was stabilized at a level of(51.51±0.016)–(51.54±0.024)U/mL,representing a 54.63%–54.72%increase compared with the original strain.The single factor test revealed that the influencing factors were time,initial pH,fermentation temperature,liquid filling amount,inoculation amount,and shaker speed.The optimal fermentation conditions were identified through an orthogonal test,which revealed the following parameters to be the most effective:24 hours,28℃,an initial pH value of 7,an inoculation amount of 5%,a rotation speed of 140 r/min,and a liquid filling volume of 100 to 250 milliliters.Under the optimal fermentation conditions,the esterase activity of the mutant strain UVDES3-2 was found to be(53.73±0.14)U/mL,representing a 61.30%increase compared to the original strain JM1-9.
作者
蔡岭肸
黄涛
但俊林
邹伟
杨海泉
CAI Lingxi;HUANG Tao;DAN Junlin;ZOU Wei;YANG Haiquan(College of Bioengineering,Sichuan University of Science&Engineering,Yibin 644005,China;Liquor Making Biotechnology and Intelligent Manufacturing of Key Laboratory of China National Light Industry,Yibin 644000,China;School of Biotechnology,Jiangnan University,Wuxi 214122,China)
出处
《食品科技》
北大核心
2024年第12期1-9,共9页
Food Science and Technology
基金
四川轻化工大学研究生创新基金项目(Y2023222)
中国轻工业酿酒生物技术及智能制造重点实验室项目(2023-05)。
关键词
酯酶
异常威客汉逊酵母
复合诱变
发酵条件优化
esterase
Wickerhamomyces anomalus
complex mutagenesis
optimization of fermentation conditions
