一株产蛋白酶植物内生菌的分离、鉴定及产酶条件研究  

Isolation, identification of a protease-producing endophytic bacteria and its culture conditions

作　　者：褚衍亮[1] 刘宇佳 姚婷 何中琼 卢波 王逸园 CHU Yanliang;LIU Yujia;YAO Ting;HE Zhongqiong;LU Bo;WANG Yiyuan(College of Biotechnology,Jiangsu University of Science and Technology,Zhenjiang 212018,China)

机构地区：[1]江苏科技大学生物技术学院,镇江212018

出　　处：《江苏科技大学学报（自然科学版）》2019年第1期93-99,120,共8页Journal of Jiangsu University of Science and Technology:Natural Science Edition

基　　金：江苏科技大学本科生创新创业训练计划项目(2017120216);江苏科技大学蚕业研究所生物技术学院科研项目(2016SY03)

摘　　要：采用明胶平板筛选法从植物防风的根中分离得到一株高产蛋白酶的内生菌株FFR3.结合生理生化特性和16S rDNA序列分析,初步鉴定此菌株为类芽孢杆菌Paenibacillus sp..以胞外蛋白酶活力为指标,对内生细菌FFR3的产酶培养条件进行初步研究.结果表明:5 g/L葡萄糖和7.5 g/L酵母浸膏是菌株生长产酶的较适碳源和氮源;无机盐MgSO_4·7H_2O、KH_2PO_4和FeSO_4·7H_2O浓度分别为3、5和0.1 g/L时有利于胞外蛋白酶的产生.菌株在发酵培养条件为2%接种量、50 mL/250 mL装液量、pH 7.0、转速180 r/m,温度37℃时,发酵48 h可获得较高的酶产量,蛋白酶活力可达633.19 U/mL,较初筛时的酶活力提高15.94倍.The endophytic strain FFR3 of a high-yield protease was isolated from the root of Radix Saposhnikoviae by gelatin plate screening method. Combined with physiological and biochemical characteristics and 16 S rDNA sequence analysis, the strain was identified as Paenibacillus sp. The enzyme culture conditions of endophytic bacteria FFR-3 were studied by using extracellular protease activity as an index. The results showed that 5 g/L glucose and 7.5 g/L yeast extract were the suitable carbon and nitrogen sources for the growth of the strain. The concentration of MgSO4·7 H2O, KH2PO4 and FeSO4·7 H2O are 3, 5 and 0.1 g/L, which is beneficial to the production of protease. The culture conditions for liquid fermentation were 2% inoculation amount, 50 mL/250 mL, pH 7.0, speed 180 r/m and temperature 37 ℃. After fermented for 48 h with these conditions, the protease activity can reach 633.19 U/mL, which was 15.9 times higher than the primary screening of enzyme activity.

关 键 词：植物内生菌 蛋白酶 发酵条件 类芽孢杆菌 

分 类 号：Q939.9[生物学—微生物学]