嗜酸乳杆菌表达载体的构建及其甘露聚糖酶的表达
Construction of Lactobacillus acidophilus expression vector and expression of mannanase
顾斌涛 1郭建军 1曾静 1聂俊辉 1王通 1熊大维 1袁林1
作者信息
- 1. 江西省科学院微生物研究所,江西南昌 330096
- 折叠
摘要
为获得产甘露聚糖酶的嗜酸乳杆菌(Lactobacillus acidophilus)菌株,该研究构建甘露聚糖酶表达载体并转化嗜酸乳杆菌,分析重组嗜酸乳杆菌产甘露聚糖酶的酶学性质,并测定重组嗜酸乳杆菌的耐酸性及甘露聚糖酶的抗蛋白酶活性.结果表明,重组嗜酸乳杆菌发酵24 h,其所产甘露聚糖酶活性为353 U/mL.甘露聚糖酶的最适催化pH值为5.5,最适催化温度为55℃.重组嗜酸乳杆菌在pH 2.0～4.5有一定的耐酸性,在pH 2.0条件下2 h的存活率为77.3％.甘露聚糖酶液用胃蛋白酶、胰蛋白酶处理160 min后,甘露聚糖酶的相对酶活分别为65％、28％.表明该甘露聚糖酶具有一定的抗胃蛋白酶和部分抗胰蛋白酶降解能力.
Abstract
In order to obtain mannanase-producing Lactobacillus acidophilus strain,the mannanase expression vector was constructed and transformed into L.acidophilus.The enzymatic properties of mannanase produced by recombinant L.acidophilus were analyzed,and the acid resistance of recom-binant L.acidophilus and anti-protease activity of mannanase were determined.The results showed that the mannanase activity produced by recombi-nant L.acidophilus was 353 U/ml after fermentation for 24 h.The optimum catalytic pH and temperature of mannanase was 5.5 and 55 ℃,respectively.The recombinant L.acidophilus had certain acid resistance at pH 2.0 to 4.5,and the survival rate was 77.3％at pH 2.0 for 2 h.After mannanase solu-tion was treated with pepsin and trypsin for 160 min,the relative activities of mannanase were 65％and 28％,respectively.The results indicated that the mannanase had certain anti-pepsin and partial anti-trypsin degradation ability.
关键词
甘露聚糖酶/嗜酸乳杆菌/质粒构建/酶学性质Key words
mannanase/Lactobacillus acidophilus/plasmid construction/enzymatic property引用本文复制引用
基金项目
江西省重点研发计划(20212BBF63022)
江西省重点研发计划(20192BBFL60020)
江西省科学院省级科技计划包干制试点示范项目(2021YSBG10005)
江西省科学院省级科技计划包干制试点示范项目(2022YSBG22006)
出版年
2024
NETL
NSTL
维普
万方数据