解淀粉芽孢杆菌SSY2颉颃霉菌物质的鉴定及其理化性质研究

doi:10.16431/j.cnki.1671-7236.2018.11.029

摘要/Abstract

摘要：

试验旨在鉴定解淀粉芽孢杆菌SSY2的抗霉菌物质及其理化性质。通过活性组分分析、硫酸铵沉淀、超滤除盐等方法鉴定并提取抗菌物质；通过对抗霉菌物质产生的时间、酶活性、温度、酸碱度（pH）、稳定性及储存温度的检测初步确定其理化性质。结果表明，不同组分对霉菌的降解率不同，菌液上清液对霉菌的降解能力最强，菌液发酵液的降解能力次之，胞内液降解能力最差。因此，初步确定解淀粉芽孢杆菌SSY2的抗菌物质活性成分是一种胞外分泌物，主要存在于菌液上清液；经30%、60%、80%和100%饱和硫酸铵沉淀，确定最佳硫酸铵饱和浓度为60%；超滤纯化后得到分子质量大于3 ku的抗菌活性物质。理化性质检测结果表明，该抗菌物质能在对数期早期（24 h）开始产生；具有产蛋白酶活性，对胃蛋白酶具有一定抵抗力，但对蛋白酶K、碱性和中性蛋白酶的抵抗力较低；在60~121℃抗菌活性稳定，证明其对热不敏感；耐酸性较强，在pH 6.0~8.0环境中对霉菌的抗菌活性稳定；此外，抗菌物质具有良好的传代稳定性，可在4℃条件下长期保存。本试验结果可为后续研发霉菌生物防控制剂提供依据。

关键词: 解淀粉芽孢杆菌; 理化性质; 霉菌

Abstract:

The test was designed to identify the antifungal substances of Bacillus amyloliquefaciens SSY2 and their physicochemical properties.Identification and extraction of antibacterial substances were carried out by active component analysis,ammonium sulfate precipitation,ultrafiltration and salt removal;The physical and chemical properties were initially determined by detecting the time,enzyme activity,temperature,pH,stability and storage temperature of the mold material.The results showed that the degradation rate of different components to mould was different.The supernatant of bacterial liquid had the strongest degradation ability to mould,the degradation ability of bacterial liquid fermentation liquid was the second,and the degradation ability of intracellular liquid was the worst.Therefore,it was preliminarily determined that the active ingredient of the antibacterial substance of Bacillus amyloliquefaciens SSY2 was an extracellular secretion,which was mainly present in the supernatant of the bacterial liquid.After 30%,60%,80% and 100% saturated ammonium sulfate precipitation,the optimum ammonium sulfate saturation concentration was determined to be 60%;After ultrafiltration purification,an antibacterial active substances with molecular mass greater than 3 ku was obtained.The results of the physical and chemical property test showed that,the active substance was produced early (24 h) in the logarithmic phase,and it had good passage stability.It had protease activity and resistance to pepsin,but it had lower resistance to protease K,alkaline and neutral protease;The antibacterial activity at 60 to 121℃ was stable,which proved that it was not sensitive to heat;There was no significant difference in the antibacterial activity of the supernatant against molds in pH 6.0 to 8.0 environment,it was proved that antimicrobial substances had strong acid resistance;Antimicrobial substances had good passage stability,and it could be stored at 4℃ for a long time.The results could provide basis for further research and development of fungal biological control agents.

Key words: Bacillus amyloliquefaciens; physicochemical property; mould

中图分类号:

S816.79

陈楠楠, 尹珺伊, 王岩, 秦平伟, 史同瑞, 刘宇. 解淀粉芽孢杆菌SSY2颉颃霉菌物质的鉴定及其理化性质研究[J]. 《中国畜牧兽医》, 2018, 45(11): 3221-3228.

CHEN Nannan, YIN Junyi, WANG Yan, QIN Pingwei, SHI Tongrui, LIU Yu. Identification and Physicochemical Properties of Antifungal Substances of Bacillus amylolyticus SSY2[J]. , 2018, 45(11): 3221-3228.

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