一株热噬淀粉芽孢杆菌的分离和鉴定

缪伏荣; 董志岩; 陈鑫珠; 刘景

doi:10.19303/j.issn.1008-0384.2018.04.015

一株热噬淀粉芽孢杆菌的分离和鉴定

doi: 10.19303/j.issn.1008-0384.2018.04.015

福建省农业科学院畜牧兽医研究所, 福建福州 350013

基金项目:

福建省科技计划项目——省属公益类科研院所基本科研专项 2015R1101

详细信息

作者简介:
缪伏荣(1969-), 男, 硕士, 副研究员, 主要从事特种微生物的研究与应用(E-mail:miaofr@sina.com)

通讯作者:
刘景(1967-), 男, 副研究员, 主要从事饲料资源的研究(E-mail:fjliuj@163.com)

中图分类号: Q93
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出版历程
- 收稿日期: 2017-10-22
- 修回日期: 2018-02-12
- 刊出日期: 2018-04-01

Isolation and Identification of Bacillus thermoamylovorans

Institute of Animal Husbandry and Veterinary Medicine, Fujian Academy of Agricultural Sciences, Fuzhou, Fujian 350013, China

摘要

摘要: 为获得在高温下能高效降解畜禽屠宰场废弃物的菌株，利用逐级梯度升温法，从畜禽屠宰场废弃物中分离筛选到菌株HL8103。对菌落形态、菌株形态、生理生化特性等进行探讨，并克隆其16S rDNA序列，进行系统发育学分析。同时进行菌株对屠宰废弃物适应性测试。结果表明，HL8103为热噬淀粉芽孢杆菌Bacillus thermoamylovorans，其形态特征为：革兰氏阳性，菌体呈杆状，直或弯曲，芽胞近圆形，近端生，孢囊膨大。该菌可耐95℃的高温，且最适生长温度为75~85℃，最适生长pH值为6.8~7.2，生长最佳时间为18 h，能适应屠宰废弃物的生长。
- 热噬淀粉芽孢杆菌 /
- 分离 /
- 鉴定
Abstract: This study aimed to identify microbes that could, at high temperatures, efficiently degrade waste materials from livestock and poultry slaughterhouses. A stepwise gradient heating was applied to screen microorganisms and isolated HL8103.Through colony morphology, strain morphology, physiological and biochemical characteristics, 16S rDNA sequence, phylogenetic analysis. At the same time, the strain was tested for its suitability for livestock and poultry slaughterhouse waste. The results identified HL8103 as Bacillus thermoamylovorans, a gram-positive with straight or curved rod-shape bacterium that had near spherical spores, proximal birth, and inflated cysts. Meanwhile, HL8103 was tested for its adaptability on the slaughterhouse waste and found that it could grow well between 75℃ and 85℃ and withstand a temperature as high as 95℃. And, for optimal results, the medium was to be adjusted to pH 6.8-7.2 and fermented for 18 h.
- Bacillus thermoamylovorans /
- separation /
- identification

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图 1 平板菌落特征

Figure 1. Characteristics of microbial colonies on petri dish

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图 2 电镜下菌株形态

Figure 2. Morphology of HL8103 under electron microscope

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图 3 HL8103菌的生长曲线

Figure 3. Growth curve of HL8103

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图 4 初始pH对HL8103菌生长的影响

Figure 4. Effect of initial pH on enzyme production of HL8103

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图 5 HL8103菌株序列系统发育树

Figure 5. Phylogenetic tree based on HL8103 sequence

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表 1 HL8103菌不同培养基中生长比较

Table 1. Growth of HL8103 on varied culture media

培养基	活菌总数/(CFU·mL^-1)
LB培养液	3.08×10⁶
20%屠宰废弃物水溶液	5.26×10⁶
灭菌20%屠宰废弃物水溶液	5.25×10⁶
注：接种后最终菌浓度均为1×10⁶CFU·mL^-1，培养时间均为18 h。

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表 2 HL8103菌生理生化特性

Table 2. Physio-biochemical characteristics of HL8103

项目	生理生化特性	项目	生理生化特性	项目	生理生化特性	项目	生理生化特性
甘油	+	赤藓糖醇	+	D-阿拉伯糖	-	L-阿拉伯糖	+
D-核糖	+	D-木糖	+	L-木糖	-	阿东醇	-
β-甲基-D-木糖苷	-	D-半乳糖	+	D-岩藻糖	-	D-果糖	+
D-甘露糖	+	L-山梨糖	+	L-鼠李糖	+	卫矛醇	-
肌醇	-	甘露醇	-	山梨糖	-	α-甲基-D-甘露糖苷	-
α-甲基-D-葡萄糖苷	+	N-乙酰葡萄糖胺	+	苦杏仁苷	+	熊果苷	-
七叶灵	+	水杨苷	+	D-纤维二糖	+	D-麦芽糖	+
D-乳糖	+	D-蜜二糖	+	D-蔗糖	-	D-海藻糖	+
菊糖	-	D-松三糖	-	D-棉籽糖	-	淀粉	+
糖原	+	木糖醇	-	D-龙胆二糖	+	D-土伦糖	+
D-来苏糖	-	D-塔格糖	+	葡萄糖酸钾	-	L-岩藻糖	-
D-阿拉伯醇	-	L-阿拉伯醇	-	葡萄糖产酸	+	2-酮基-葡萄糖酸盐	-
5-酮基-葡萄糖酸盐	-	吲哚	-	葡萄糖产气	+	酵母粉	+
胰蛋白胨	+	硝酸盐	+	铵盐	+
注：+为阳性，-为阴性。

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