ARTP-DES连续诱变选育高产ε-聚赖氨酸突变株  

Sequential Mutagenesis Treatment with Atmospheric and Room Temperature Plasma and Diethyl Sulfate for Enhanced Microbial Production of ε-Poly-L-lysine

作　　者：席志文 黄林娜[1] 翟一畅 惠丰立[1] XI Zhiwen;HUANG Linna;ZHAI Yichang;HUI Fengli(College of Life Science and Technology,Nanyang Normal University,Nanyang 473061,China)

机构地区：[1]南阳师范学院生命科学与技术学院,河南南阳473061

出　　处：《食品科学》2020年第14期131-137,共7页Food Science

基　　金：国家自然科学基金面上项目(31570021)。

摘　　要：为提升小白链霉菌(Streptomyces albluus)产ε-聚赖氨酸(ε-poly-L-lysine,ε-PL)能力,在常压室温等离子体(atmospheric and room temperature plasma,ARTP)、硫酸二乙酯(diethyl sulfate,DES)单因子诱变的基础上,采用ARTP-DES连续诱变。连续诱变条件:ARTP工作功率110 W,照射距离2 mm,照射时间为30、60、105 s;DES诱变体积分数0.6%,反应时间20 min。并改进一种快速筛选方法,结合链霉素抗性、亚甲基蓝透明圈及48微孔板培养,酶标仪快速测定ε-PL含量。结果表明:ARTP-DES连续诱变结合链霉素抗性正突变率可达40.8%,同时,获得1株遗传性能稳定的链霉素抗性突变株S. albulus AD-9,其ε-PL摇瓶产量为2.1 g/L,是出发菌株的2.1倍。通过研究高产菌株的生理特性,发现其在营养需求、发酵过程中菌球形态等都发生了变化。ARTP-DES连续诱变选育高产ε-PL菌株是一种高效选育手段。To enhance the production of ε-poly-L-lysine(ε-PL) by Streptomyces albluus, we mutagenized this strain by sequential treatment with atmospheric and room temperature plasma and diethyl sulfate(ARTP-DES). The optimized treatment conditions were determined as follows: ARTP power 110 W, exposure distance 2 mm, mutagenesis time 30, 60, and 105 s, DES concentration 0.6%(V/V), and reaction time 20 min. A rapid screening method was established by streptomycin resistance combined with methylene blue transparent circles. ε-PL content was determined by enzyme-linked immunosorbent assay(ELISA) with a 48-well microplate. The results showed that the positive mutation rate for streptomycin resistance was up to 40.8%. Furthermore, one strain designated AD-9 with good genetic stability was selected. It produced 2.1 g/L ε-PL in shake-flask fermentation, which was 2.1 times as high as that produced by the parent strain. Studies of the physiological characteristics indicated that the mutant strain AD-9 showed changes in nutritional requirements, pellet morphology and nutrient consumption during fermentation. This study indicates that continuous ARTP-DES mutagenesis is an efficient breeding method for increased production of ε-PL.

关 键 词：小白链霉菌 Ε-聚赖氨酸 连续诱变 快速筛选 生理特性 

分 类 号：TQ920.1[轻工技术与工程—发酵工程]