耐碱性β甘露聚糖酶产生菌的分离鉴定及发酵条件优化

应用潜力。
关键词：耐碱β-甘露聚糖酶；筛选；鉴定；发酵条件优化；酶学性质
doi:10.13302/ki.fi.2019.05.005
中图分类号：S816.32
文献标识码：A
文章编号：1001-991X（2019）05-0023-11
Isolation, identification and optimization of fermentation conditions of an alkali-resistant
potential for application. Key words：alkali-resistant β-Mannanase；screen⁃ ing；identification；fermentation condition optimi⁃ zation；enzymatic property
10.0 g/l、CaCl2 0.6 g/l、K2HPO4 0.8 g/l、MgSO4·7H2O 1.0 g/l，pH 值 7.51；接种量 2%，装液量 40 ml/250 ml 三角瓶，37 ℃、220 r/min 发酵 36 h，酶活力达 355.75 U/ml，是初始酶活力的 3.5 倍。该酶的最适反
lus sp. and optimization its culture conditions. A highly β- Mannanase producing Bacillus subtilis
named WMYB- 2, was screened by transparent circle method. Single- factor experiment and Box-
摘 要：研究旨在高产β-甘露聚糖酶饲用益生芽孢杆菌的筛选及培养条件优化。用透明圈法筛
选得到一株产β-甘露聚糖酶活力较高的枯草芽孢杆菌（Bacillus subtilis）WMYB-2，通过单因素实验、
Box-Behnken 实验及响应面分析对该菌的产酶培养基及培养条件进行了优化，并对其酶学性质进行
了初步研究。结果表明，该菌株产酶的最佳培养基及发酵条件为：魔芋微粉 10.0 g/l，大豆蛋白胨
应温度和 pH 值分别为 55 ℃和 5.5。在 45~55 ℃内酶活力稳定，55 ℃保温 30 min 和 3 h 后其相对
酶活力保留 92%和 57%；在 pH 值 5.0~10.0 内酶活力稳定，在 pH 值 9.0 和 10.0 的环境下处理 2 h 其
相对酶活力保留 95%和 81%。Li+和 K+对该酶具有一定的激活作用。该酶具有良好的耐碱特性及
57% after a treatment of 3 hours. The enzyme activity remained 95% and 81% after a treatment of 2
hours at pH 9.0 and 10.0. The enzyme was acti⁃
vated by Li+ and K+. The enzyme showed excel⁃
perature and pH of the enzyme were 55 ℃ and 5.5. The enzyme was stable between temperature 45~
55 ℃ and pH 5.0~10.0. The enzyme activity remained 92% at 55 ℃ for a treatment of 30 minutes and
2019 年第 40 卷第 5 期 总第 578 期
SILIAO GONGYE
耐碱性β - 甘露聚糖酶产Байду номын сангаас菌的分离鉴定 及发酵条件优化
■ 汪梦昀 1 缪礼鸿 1* 励 飞 2 刘蒲临 1 廖卫芳 1
（1.武汉轻工大学生物与制药工程学院，湖北武汉 430023；2.湖南九鼎科技（集团）有限公司，湖南长沙 410000）
作者简介 ：汪梦昀 ，硕士 ，研究方向为资源微生物。 通讯作者 ：缪礼鸿 ，博士 ，教授。 收稿日期 ：2019-01-08 基 金 项 目 ：国 家“863”高 技 术 研 究 发 展 计 划 项 目 [2013AA102805]
lent alkali resistance characteristics, and had the
sult showed that culture with an inoculation size of 2% for 36 h at 37 ℃ and 220 r/min in a 250 ml
shaking flask filled with 40 ml liquid medium consisting of 10.0 g/l konjac powder, 10.0 g/l soybean
peptone, CaCl2 0.6 g/l, K2HPO4 0.8 g/l, MgSO4·7H2O 1.0 g/l, at initial pH 7.51, the β-Mannanase ac⁃
tivity reached 355.75 U/ml, which was 3.5 times higher than before the optimization. The optimum tem⁃
Behnken experiment of response surface analysis were performed to obtain the optimal medium and fer⁃
mentation conditions for β-Mannanase production, and the enzymatic properties were studied. The re⁃
β-Mannanase producing strain
Wang Mengyun, Miao Lihong, Li Fei, Liu Pulin, Liao Weifang
Abstract：The experiment was conducted to screen the highly β-Mannanase producing probiotic Bacil⁃