食品工业科技2024,Vol.45Issue(4) :101-108.DOI:10.13386/j.issn1002-0306.2023040051

高产抗菌脂肽芽孢杆菌的分离鉴定筛选及发酵工艺优化

Isolation,Identification,Screening and Fermentation Process Optimization of Bacillus Producing High Antimicrobial Lipopeptide

纪帅奇 乌日娜 张淘崴 娄梦雪 贺凯茹 张妍 武俊瑞
食品工业科技2024,Vol.45Issue(4) :101-108.DOI:10.13386/j.issn1002-0306.2023040051

高产抗菌脂肽芽孢杆菌的分离鉴定筛选及发酵工艺优化

Isolation,Identification,Screening and Fermentation Process Optimization of Bacillus Producing High Antimicrobial Lipopeptide

纪帅奇 1乌日娜 2张淘崴 3娄梦雪 4贺凯茹 3张妍 1武俊瑞2
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作者信息

  • 1. 沈阳农业大学食品学院,辽宁沈阳 110866;辽宁省食品发酵技术工程研究中心,辽宁沈阳 110866
  • 2. 沈阳农业大学食品学院,辽宁沈阳 110866;辽宁省食品发酵技术工程研究中心,辽宁沈阳 110866;沈阳市微生物发酵技术创新重点实验室,辽宁沈阳 110866
  • 3. 辽宁省食品发酵技术工程研究中心,辽宁沈阳 110866;沈阳市微生物发酵技术创新重点实验室,辽宁沈阳 110866
  • 4. 沈阳农业大学食品学院,辽宁沈阳 110866;沈阳市微生物发酵技术创新重点实验室,辽宁沈阳 110866
  • 折叠

摘要

为了筛选出高产抗菌脂肽的芽孢杆菌并确定其最佳发酵工艺条件,研究从传统发酵豆酱中分离和鉴定产抗菌脂肽芽孢杆菌菌株,通过正交试验研究了发酵接种量、发酵填液量、发酵时间、发酵温度等发酵条件对芽孢杆菌脂肽产量的影响.结果表明,从 9份东北传统发酵豆酱中分离筛选得到 27株芽孢杆菌,经 16S rDNA鉴定,其中 6株芽孢杆菌具有合成脂肽的基因sfp、fenB和ituA.通过单位菌体脂肽产量和抑菌效果测定发现,贝莱斯芽孢杆菌SN-20和解淀粉芽孢杆菌SN-46表现优异,单位菌体脂肽产量达 106和 72 mg,且对革兰氏阳性和阴性指示菌均有较强的抑菌效果.正交试验发现贝莱斯芽孢杆菌SN-20最佳发酵工艺为:接种量 3%,发酵填液量 20%,发酵时间36 h,发酵温度32℃;枯草芽孢杆菌SN-46最佳发酵工艺为:接种量2%,发酵填液量40%,发酵时间24 h,发酵温度 32℃.在此条件下发酵的两株芽孢杆菌单位菌体脂肽产量优化前为 106.11和 76.23 mg/g,优化后分别提高了21.85%和23.84%.研究结果有效地增加了芽孢杆菌抗菌脂肽的产量.

Abstract

In order to screen out Bacillus sp.with high production of antimicrobial lipopeptide and determine its optimal fermentation conditions,the study isolated and identified antimicrobial lipopeptide-producing Bacillus sp.strains from traditional fermented soybean paste,and adopted orthogonal test to investigate how lipopeptide production by Bacillus sp was affected by fermentation conditions,such as fermentation inoculum amount,fermentation filling volume,fermentation time and fermentation temperature.The results showed that 27 strains of Bacillus sp.were isolated and screened out from 9 portions traditional fermented soybean paste of the Northeast,among which 6 strains were identified by 16S rDNA as having the genes sfp,fenB and ituA for synthesizing lipopeptide.According to the determination of lipopeptide yield and inhibition effect,Bacillus subtilis SN-20 and Bacillus amyloliquefaciens SN-46 showed excellent performance with the yield of 106 and 72 mg of lipopeptide per unit,and strong inhibition effect on both gram-positive and negative indicator bacteria.The optimal fermentation process of Bacillus subtilis SN-20 was found to be 3%inoculum,20%fermentation filler,36 h fermentation time and 32℃.The optimal fermentation process of Bacillus subtilis SN-46 was 2%inoculum,40%fermentation filler,24 h fermentation time and 32℃.Under these conditions,the unit biomass yields of lipopeptides for the two strains of Bacillus before optimization were 106.11 and 76.23 mg/g,respectively.After optimization,they increased by 21.85%and 23.84%,respectively.The study results effectively increased the production of antimicrobial lipopeptides from Bacillus.

关键词

抗菌脂肽/芽孢杆菌/筛选/鉴定/发酵工艺优化

Key words

antimicrobial lipopeptide/Bacillus/screening/characterization/optimization of fermentation process

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基金项目

国家自然科学基金面上项目(32172279)

沈阳市科技创新平台项目(21-103-0-14)

沈阳市科技创新平台项目(21-104-0-28)

出版年

2024
食品工业科技
北京一轻研究院

食品工业科技

CSTPCD北大核心
影响因子:0.842
ISSN:1002-0306
参考文献量16
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