一株解淀粉芽胞杆菌的筛选及其益生特性研究

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摘要摘要芽胞杆菌(Bacillus)已被广泛用作动物益生菌，其分泌的纤维素酶(cellulose)可以提高饲料利用率，改善动物肠道微生物菌群结构。本研究利用羧甲基纤维素钠(sodium carboxymethyl cellulose, CMC-Na)培养基从甘蔗(Saccharum officinarum)、麻类等纤维作物根际分离具有较高纤维素酶活性的芽胞杆菌，并对其纤维素酶的产酶条件、动物肠胃耐受性、对滤纸条纤维素的降解情况以及菌株的溶血性检验安全性进行鉴定。研究结果表明，获得了8株具有产纤维素酶活性的芽胞杆菌，其中菌株FJAT-29941的活性最高，形态学、生理生化和16S rRNA基因鉴定结果显示，该菌株为解淀粉芽胞杆菌(Bacillus amyloliquefaciens)；菌株FJAT-29941产纤维素酶的培养温度为20~50 ℃(最佳培养温度为30 ℃)，pH为4.0~9.0(最佳pH值为9)，培养时间18~72 h(最佳培养时间为42 h)，最佳培养基为0.5% NaCl的营养琼脂(nutrient agar, NA)培养基；利用1%胰蛋白酶的人工肠液和1%胃蛋白酶的人工胃液处理该菌株，其存活率分别为27.98%和64.91%，表明其具有较好的肠胃耐受性；菌株FJAT-29941在48 h内可以将滤纸条降解为小圆片，表现出较强的纤维素降解能力；该菌株在溶血性检测中未产生透明圈，是安全可应用的。本研究筛选获得的解淀粉芽胞杆菌FJAT-29941具有较好的纤维降解能力和动物肠胃耐受性，为益生菌研发提供了菌种资源和科学依据，具有良好的开发前景。

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	刘国红
	刘波
	车建美
	王阶平
	陈倩倩
	姚运发
	刘琴英

关键词 ：解淀粉芽胞杆菌, 纤维素酶, 肠胃耐受性, 益生菌

Abstract：Abstract Bacillus strains are widely used as animal probiotics. Cellulase produced by Bacillus strains could be used to increase efficiency of feed utilization and improve the intestinal microflora. The aim of this study was to screen the Bacillus strains with high celluloytic activities from the rhizosphere soil of plants (sugarcane and hemp, and so on) to provide strain resources for development of animal probiotics. Firstly, culturable Bacillus strains were isolated from the soil samples through dilution seperation method on LB plate. Eight celluloytic Bacillus strains were obtained by screening with the sodium carboxymethyl cellulose (CMC-Na) medium. Among them, the strain FJAT-29941 exhibited the strongest cellulase activities and was identified as Bacillus amyloliquefaciens according to the morphology, physical and biochemical characteristics and 16S rRNA gene sequences analysis. The cellulase production conditions, gastrointestinal tolerance, filter paper degradation and hemolytic activity of strain FJAT-29941 were studied furtherly. The fermentative conditions for cellulase-producing of the strain FJAT-29941 were as follows: pH 4.0~9.0 (optimum pH 9.0), 20~50 ℃ (optimum 30 ℃ ), 18~72 h (optimum 42 h) and the optical fermentative medium was NA (nutrient agar) containing 0.5% NaCl. Moreover, the cellulases displayed excellent tolerance to pH and temperature. The survival rate of the strain FJAT-29941 in the artificial intestinal fluid containing 1% typsin and the artificial gastric fluid containing 1% pepsin was 27.98% and 64.91%, respectively, suggesting a great tolerance to gastrointestinal environments. It was showed that the cultures could rapidly degrade intact long filter paper scrips into fragments within 48 hours of incubation, showing strong ability of degradation on cellulose substrates. Finally, no hemolysis of the strain FJAT-29941 was observed on the goat blood agar plates, indicating its safety toward use of this isolate as a potential probiotics. Taken together, the strain B. amyloliquefaciens FJAT-29941 exhibited both excellent cellulose degrading ability and probiotic potential, which would provide a strain resource and scientific basis for the probiotics development.

Key words： Bacillus amyloliquefaciens Cellulose Gastrointestinal tolerance Probiotic

收稿日期: 2017-11-13 出版日期: 2018-05-21

基金资助:福建省自然科学基金;福建省公益项目;农业部农业微生物资源收集与保藏重点实验室开放基金;农业部948研究项目

通讯作者: 刘波 E-mail: liubofaas@163.com;fzliubo@163.com

引用本文:

刘国红刘波车建美王阶平陈倩倩姚运发刘琴英. 一株解淀粉芽胞杆菌的筛选及其益生特性研究[J]. , 2018, 26(6): 1043-1055.

链接本文:

http://journal05.magtech.org.cn/Jwk_ny/CN/ 或 http://journal05.magtech.org.cn/Jwk_ny/CN/Y2018/V26/I6/1043

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