ε-聚赖氨酸是一种典型的同型L-赖氨酸聚合物,具有广谱抗菌活性和抗噬菌体活性,广泛应用于食品和医药行业。该文克隆了经密码子优化的白色链霉菌Streptomyces albulus来源的ε-聚赖氨酸合酶编码基因pls,首次实现了其在食品安全性菌株枯草芽孢杆菌Bacillus subtilis 168中的异源表达。其次,对B. subtilis 168/pMA5-pls重组菌株生产ε-聚赖氨酸的全细胞催化体系进行了优化,结果表明,在L-赖氨酸质量浓度0.5 g/L, 初始pH和温度分别为3.0和30 ℃条件下,ε-聚赖氨酸合酶的转化效果最好。在最优体系条件下,连续转化4 h,ε-聚赖氨酸的产量达到195.1 mg/L,高于已报道的Bacillus subtilis直接发酵生产的ε-聚赖氨酸产量。该研究构建的全细胞催化体系为食品级ε-聚赖氨酸的生产提供了新策略。
ε-poly-L-lysine is a typical L-lysine homopolypeptide with broad-spectrum antibacterial activity and anti-phage activity, which is widely used in food and pharmaceutical industries. Firstly, a codon optimized pls gene, encoding ε-poly-L-lysine synthetase from Streptomyces albulus was cloned, and for the first time realized its heterologous expression in GRAS strain Bacillus subtilis 168. Secondly, the whole-cell catalytic system for the production of ε-poly-L-lysine by the recombinant strain B. subtilis 168/pMA5-pls were optimized. The results showed that at a concentration of 0.5 g/L L-lysine, initial pH and temperature of 3.0 and 30 ℃ respectively, the conversion effect of ε-poly-L-lysine synthetase was best. Finally, under the optimal condition, continuous conversion of 4 hours, yielded approximately 195.1 mg/L of ε-poly-L-lysine, which was higher than the reported yield of ε-poly-L-lysine produced by Bacillus subtilis via direct fermentation. Therefore, the whole-cell catalytic system constructed in this study provides a new strategy for the safe production of ε-poly-L-lysine.
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