食品与发酵工业 >

2021 , Vol. 47 >Issue 17: 84 - 90

DOI: https://doi.org/10.13995/j.cnki.11-1802/ts.026262

研究报告

高产中性蛋白酶菌株的诱变选育及益生特性

  • 黄子凌 ,
  • 莫港澳 ,
  • 李文 ,
  • 王陶
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  • 1(徐州工程学院,江苏 徐州,221018)
    2(江苏省食品资源开发与质量安全重点建设实验室,江苏 徐州,221018)
本科生(王陶教授为通讯作者,E-mail:wangtaohf@126.com)

收稿日期: 2020-11-24

  修回日期: 2020-12-25

  网络出版日期: 2021-09-27

基金资助

江苏省政策引导类计划项目(苏北科技专项)(XZ-SZ201821);江苏省“六大人才高峰”高层次人才项目(SWYY-222);江苏省大学生创新创业训练计划项目(XCX2020048);徐州工程学院大学生创新创业基金项目(2019030)

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Probiotic characteristics and mutagenesis breeding of neutral protease high-yield strain

  • HUANG Ziling ,
  • MO Gangao ,
  • LI Wen ,
  • WANG Tao
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  • 1(Xuzhou University of Technology,Xuzhou 221018,China)
    2(Jiangsu Key Construction Laboratory of Food Resource Development and Quality Safe,Xuzhou 221018,China)

Received date: 2020-11-24

  Revised date: 2020-12-25

  Online published: 2021-09-27

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摘要

以产中性蛋白酶的枯草芽胞杆菌WT为出发菌株,通过理化诱变来筛选高产菌株,并检测高产菌株耐热、耐模拟胃肠液、表面特性、降脂能力和对几种致病菌的抑菌能力来评价其益生特性。以紫外、LiCl为诱变剂,通过3轮诱变后,得到了1株能稳定高产中性蛋白酶的突变菌株UL-191,其中性蛋白酶活力较出发菌株WT提高了44.2%,达108.812 U/mL。UL-191具有较好的耐热性,90 ℃处理后芽孢存活率能达到66.87%;对于模拟胃、肠液UL-191的耐受性较好,3 h后存活率能分别达到78.23%、60.00%;降胆固醇和甘油三酯的能力都较好,分别可达35.00%和49.12%;对于菌株表面特性而言,其表面凝聚性能达到69.38%,以二甲苯为有机相时的表面疏水性可达46.02%,以正己烷为有机相时的表面疏水性可达31.41%;对大肠杆菌、沙门氏菌和金黄色葡萄球菌的抑制能力较强,抑菌圈直径分别可达14.90、17.52和23.40 mm。突变菌株UL-191能高产中性蛋白酶,又具有良好的益生特性,在食品、饲料等行业具有较好的应用前景。

关键词： 枯草芽胞杆菌; 中性蛋白酶; 理化诱变; 益生特性

本文引用格式

黄子凌 , 莫港澳 , 李文 , 王陶 . 高产中性蛋白酶菌株的诱变选育及益生特性[J]. 食品与发酵工业, 2021 , 47(17) : 84 -90 . DOI: 10.13995/j.cnki.11-1802/ts.026262

Abstract

Bacillus subtilis WT was mutagenized by ultraviolet irradiation and lithium chloride to obtain high-yield neutral protease strain. The probiotic characteristics were evaluated by heat tolerance, simulated gastric and intestinal fluid tolerance, cell-surface property, degradation activity of cholesterol and triglyceride, and antibacterial activity. Mutant UL-191 with stable and high yield of neutral protease was obtained. The activity of neutral protease was 108.812 U/mL, which increased by 44.2% compared with WT. UL-191 had strong heat and simulated gastrointestinal fluid resistance, the survival rates was 66.87% at 90 ℃ and reached 78.23%, 60.00% after 3 hours. Furthermore, UL-191 could degrade cholesterol and triglyceride with degradation rates of 35.00% and 49.12%, respectively. The cell surface cohesion was 69.38%, the surface hydrophobicity were 46.02% and 31.41% when the organic phase were xylene and hexane, respectively; the strain could inhibited the growth of Escherichia coli, Staphylococcus aureus, Salmonella typhimurium with the inhibition zone reaching to 14.90, 17.52 and 23.40 mm. In summary, mutant UL-191 could produce neutral protease with high-yield, and showed good probiotic characteristics, thus it could have a good application prospect in food, feed and other industries.

Key words： Bacillus subtilis; neutral protease; physical and chemical mutagenesis; probiotic characteristics

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