产蛋白酶海洋细菌Bacillus sp. 4sp5分离鉴定及降解南极磷虾原肌球蛋白的研究

Isolation and characterization of the protease-producing marine bacterium Bacillus sp. 4sp5 and its degradation of Antarctic krill tropomyosin

中文关键词:  原肌球蛋白  南极磷虾  海洋细菌  蛋白酶  脱敏作用

英文关键词:tropomyosin  Antarctic krill  marine bacteria  protease  allergy removal

中文摘要:

      目的 从海洋样品中筛选产蛋白酶海洋细菌及可以降解南极磷虾原肌球蛋白(tropomyosin, TM)的蛋白酶。方法 采用透明圈法筛选产蛋白酶菌株, 并根据聚丙烯酰胺凝胶电泳(polyacrylamide gel electrophoresis, PAGE)和酶联免疫吸附测定法(enzyme-linked immunosorbent assay, ELISA)确定降解效果最佳菌株, 对其产的蛋白酶的酶学特性及降解原肌球蛋白条件进行研究。结果 最佳为菌株4sp5, 经16S rDNA鉴定为Bacillus sp.。蛋白酶的最适作用温度和pH分别为60℃和9。Na+和K+对蛋白酶活有促进作用, 其对有机溶剂敏感, 表面活性剂对其影响较小。3种抑制剂均可抑制酶的活性。当加酶量为24 U/mL时, 7.0 h能够有效地降解南极磷虾原肌球蛋白, 经ELISA试剂盒测定, 其致敏性消除率达98.63%。结论 菌株4sp5及其产生的蛋白酶在南极磷虾原肌球蛋白脱敏作用中具有应用前景。

英文摘要:

      Objective To screen protease-producing marine bacteria produced protaese which could degrade tropomyosin (TM) of krill Antarctic from marine samples Methods The protease-producing strains were screened by the transparent circle method, and the ability of hydrolyzed TM was determined by polyacrylamide gel electrophoresis (PAGE) and enzyme-linked immunosorbent assay (ELISA). The enzymatic characteristics of the protease and the degradation conditions of tropomyosin were studied. Results The best strain 4sp5 was identified as Bacillus sp. by 16S rDNA. The optimal temperature and pH of the protease were 60℃ and 9, respectively. Na+ and K+ promoted the activity of the protease, which was sensitive to organic solvents, and had little effect on surfactants. The 3 kinds of inhibitors inhibited the activity of the protease. When the enzyme dosage was 24 U/mL, it could effectively degrade Antarctic krill myosin for 7.0 h. The sensitivity elimination rate of the enzyme was determined by ELISA kit to reach 98.63%. Conclusion The protease produced by strain 4sp5 has application prospects in the desensitization of tropomyosin in krill.