北黄海沉积物可培养产蛋白酶细菌分离鉴定

doi:DOI: 10.13343/j.cnki.wsxb.20160452

YIC-IR > 海岸带生物学与生物资源利用重点实验室 > 海岸带生物学与生物资源保护实验室

	北黄海沉积物可培养产蛋白酶细菌分离鉴定
其他题名	Cultivable protease-producing bacteria in the sediments of North Yellow Sea
	伍朝亚 ; 李岩 ; 曲慧敏 ; 张振鹏 ; 解志红; 孟宪刚
发表期刊	微生物学报
ISSN	0001-6209
	2017-04-14
卷号	57 期号:10 页码:1504-1516
关键词	marine sediment protease-producing bacteria diversity serine protease North Yellow Sea 海洋沉积物产蛋白酶细菌多样性丝氨酸蛋白酶北黄海
DOI	DOI: 10.13343/j.cnki.wsxb.20160452
产权排序	(1) 兰州交通大学化学与生物工程学院; (2) 中国科学院烟台海岸带研究所; (3) 烟台大学海洋学院
作者部门	海岸带生物学与生物资源保护实验室
英文摘要	[Objective] To uncover the diversity of extracellular protease-producing bacteria and to expand our knowledge on protease-producing bacteria in the sediments of North Yellow Sea, and to screen efficient protease production strains that might provide flora resources for mining marine protease-producing microorganisms. [Methods] Protease-producing bacteria were isolated by using casein gelatin plate from 5 sediment samples of the North Yellow Sea. The bacteria diversity was evaluated through phylogenetic analyses based on 16S rRNA genes. The protease diversity was evaluated through the inhibition tests for 39 strains with higher enzymatic activities by using 4 inhibitors: phenyl methyl sulfonyl fluoride (PMSF, serine protease inhibitor), 1,10-phenanthroline (O-P, metalloproteinase inhibitor), E-64 (cysteine protease inhibitor) and pepstatin A (aspartic protease inhibitor). Furthermore, degradation abilities for different protein substrates such as casein, gelatin and elastin were evaluated through observing hydrolytic zones. [Results] A total of 66 protease-producing strains were isolated from 5 sediment samples. These isolates were classified into 7 genera of 4 phyla including Bacteroidetes, Proteobacteria, Actinobacteria and Firmicutes, with Pseudoalteromonas (69.9%), Sulfitobacter (12.1%) and Salegentibacter (10.6%) as the dominant. The richness of the cultivable protease-producing bacteria reached 10~4 cells/g in all sediment samples. The inhibition tests indicated that all the tested strains produced serine protease and/or metal protease, only a few strains produced cysteine protease or aspartic protease. [Conclusion] The cultivable protease-producing bacteria in the North Yellow Sea are diverse with Pseudoalteromonas, Sulfitobacter and Salegentibacterbacteria as the dominant groups and the extracellular proteases belong to serine proteases and/or metalloproteinases.
中文摘要	【目的】揭示北黄海沉积物中可培养产胞外蛋白酶细菌及蛋白酶多样性,增加人们对北黄海生态系统中产蛋白酶菌多样性的认识,为海洋产蛋白酶微生物的挖掘提供菌群资源。【方法】分别将5个北黄海沉积物样品梯度稀释涂布至酪蛋白明胶筛选平板,选择性分离产蛋白酶细菌;并通过分析基于16S rRNA基因序列的系统发育关系,揭示这些细菌的分类地位和遗传多样性;分别测定胞外蛋白酶活性并对酶活较高的39株菌进行基于苯甲基磺酰氟(PMSF,丝氨酸蛋白酶抑制剂)、邻菲罗啉(o-phenanthroline,O-P,金属蛋白酶抑制剂)、E-64 (半胱氨酸蛋白酶抑制剂)和pepstatin A (天冬氨酸蛋白酶抑制剂) 4种抑制剂的酶活抑制实验以及所有菌株对3种底物(酪蛋白、明胶、弹性蛋白)的水解能力;分析这些细菌所产胞外蛋白酶的特性及多样性。【结果】从5个北黄海沉积物样品中分离获得66株产蛋白酶细菌,这些菌株隶属于Bacteroidetes、Proteobacteria、Actinobacteria和Firmicutes 4个门的7个属,其中Pseudoalteromonas (69.9%)、Sulfitobacter (12.1%)和Salegentibacter (10.6%)是优势菌群;沉积物中可培养的产蛋白酶细菌的丰度为104 CFU/g;蛋白酶酶活抑制实验表明所有测定菌株产生的胞外蛋白酶属于丝氨酸蛋白酶和/或金属蛋白酶,仅有少数菌株所产蛋白酶具有半胱氨酸蛋白酶或天冬氨酸蛋白酶活性。【结论】北黄海沉积物中可培养产蛋白酶细菌类群较为丰富,Pseudoalteromonas、Sulfitobacter和Salegentibacter菌株是优势菌群,测定菌株所产胞外蛋白酶主要是丝氨酸蛋白酶和/或金属蛋白酶。
文章类型	Article
收录类别	CSCD
语种	中文
关键词[WOS]	Microbiology
研究领域[WOS]	Microbiology ; 微生物学
CSCD记录号	CSCD:6077614
引用统计
文献类型	期刊论文
条目标识符	http://ir.yic.ac.cn/handle/133337/24245
专题	海岸带生物学与生物资源利用重点实验室_海岸带生物学与生物资源保护实验室
作者单位	1.兰州交通大学化学与生物工程学院; 2.中国科学院烟台海岸带研究所; 3.烟台大学海洋学院
推荐引用方式 GB/T 7714	伍朝亚,李岩,曲慧敏,等. 北黄海沉积物可培养产蛋白酶细菌分离鉴定[J]. 微生物学报,2017,57(10):1504-1516.
APA	伍朝亚,李岩,曲慧敏,张振鹏,解志红,&孟宪刚.(2017).北黄海沉积物可培养产蛋白酶细菌分离鉴定.微生物学报,57(10),1504-1516.
MLA	伍朝亚,et al."北黄海沉积物可培养产蛋白酶细菌分离鉴定".微生物学报 57.10(2017):1504-1516.