虾酱中益生芽孢杆菌的筛选及其潜在益生效果的研究  

Screening and Analysis of Potential Probiotic Effects of Bacillus from Shrimp Paste

作　　者：田亮金 朱薇薇 蔡岩[1,2] 张冬冬 姜柳 张倩 关博元 王世锋[1,2] Tian Liangjin;Zhu Weiwei;Cai Yan;Zhang Dongdong;Jiang Liu;Zhang Qian;Guan Boyuan;Wang Shifeng(State Key Laboratory of Marine Resource Utilization in South China Sea,College of Marine Science,Hainan University,Haikou 570228,China;Key Laboratory of Tropical Hydrobiology and Biotechnology,Hainan University,Haikou 570228,China)

机构地区：[1]海南大学南海海洋资源利用国家重点实验室,海南海口570228 [2]海南大学海洋学院,海南省热带水生生物技术重点实验室,海南海口570228

出　　处：《海南大学学报（自然科学版）》2021年第3期269-279,共11页Natural Science Journal of Hainan University

基　　金：海南省重点研发计划(ZDYF2019044)。

摘　　要：为了获得具有潜在促生长及拮抗病原菌能力的水产动物益生菌,本研究在分离和筛选的基础上,从虾酱中获得6株菌,经测序和分子鉴定后,进一步评估了其耐人工胃肠液性能、自凝集率、表面疏水活性和抗生素敏感性.结果表明:这6株菌均产酸和不溶血,其中A20、B3、B47和B89的产酸能力较好,可使发酵液pH降低1.5;A20、A27、A35、B47的蛋白酶和淀粉酶活性显著高于其他菌株,水解圈大小在4 mm以上;A20和B3对5种鱼类致病菌均具有较强的拮抗能力,抗菌圈直径H(mm)和菌落直径C(mm)的比值H/C超过1.5;16S rRNA测序鉴定表明,A20、A27、B3和B89为巨大芽孢杆菌(Bacillus megaterium),A35和B47则分别为枯草芽孢杆菌(B.subtilis)和弯曲芽孢杆菌(B.flexus).进一步的实验表明:A20、A27、B47和B89具有较强的耐人工胃液能力,菌株存活率在60%左右,A27在肠液处理后的存活率高达90%;A20和A27于各个时间段的自凝集率都高于其他菌株,24 h时达到了98%;A35和B89在三种溶剂中都表现出较高的疏水活性;A20对21种抗生素敏感,敏感率最高为80.7%.综合以上结果,巨大芽孢杆菌A20具有优良的益生潜能,可将其用于后续益生菌制剂的开发和利用.In order to obtain the probiotics of aquatic animals with potential growth promoting and antagonistic abilities,in our report,six strains were isolated and identified from shrimp paste by measuring their acid producing,hemolysis,enzyme producing and antagonistic abilities,and the molecular identification,the resistance to artificial gastrointestinal fluid,self-agglutination,hydrophobic activity and antibiotic sensitivity of the six isolates were further evaluated.The results showed all six isolates are acid producing and non-hemolytic,among which A20,B3,B47 and B89 show the best acid producing ability,reducing the pH of fermentation broth by about 1.5;the protease and amylase activities of A20,A27,A35 and B47 are significantly higher than that of the other isolates,with hydrolysis circles of more than 4 mm in diameter;A20 and B3 displaye strong antagonistic ability against five pathogenic bacteria,whose H/C ratios were both above 1.5;molecular identification based on 16 S rRNA sequencing identify A20,A27,B3 and B89 as Bacillus megaterium,A35 and B47 as B.subtilis and B.flexus,respectively.The further analyses indicated that A20,A27,B47,and B89 have strong resistance to artificial gastric fluid,their survival rates are about 60%;the survival rate of A27 is as high as 90%after intestinal fluid treatment;A20 and A27 has higher self-agglutination than the other strains at any given time period,both reaching 98%at 24 hours;A35 and B89 show high hydrophobic activities in all three solvents;A20 has the highest sensitive rate(80.7%,sensitive to 21 antibiotics)among all strains.These data suggested that B.megaterium A20 has great probiotic potential to be further studied in the subsequent development and utilization of the probiotics of aquatic animals.

关 键 词：虾酱 益生菌 芽孢杆菌 产酶能力 病原菌拮抗 

分 类 号：S963.73[农业科学—水产养殖]