传统锦州虾酱中产蛋白酶嗜盐菌的分离与鉴定

Isolation and identification of protease- producing halophilic bacteria in traditional Jinzhou shrimp paste

目的：从传统锦州虾酱中分离产蛋白酶嗜盐菌，并对其进行鉴定。方法：应用1．0％脱脂乳Gibbons培养基分离产蛋白酶嗜盐菌，经形态学、生理生化和分子生物学对其鉴定。采用SDS-PAGE分析虾酱中可溶性蛋白的变化。结果：获得16株产蛋白酶嗜盐菌，菌株CW0-1、CW0-2、CW0-3和CW0-4是弧菌属，菌株CW1-1、CW2-1和CW2-2是尼泊尔葡萄球菌，菌株CW2-3、CW5-1、CW5-2、CW5-3是马胃葡萄球菌，菌株CW3-1、CW3-2、CW4-1、CW4-2是枝芽孢杆菌，菌株CW4-3是独岛枝芽孢杆菌。弧菌属主要存在于原料虾中，嗜盐性葡萄球菌出现在发酵2个月和5个月，枝芽孢杆菌出现在发酵3～4个月。葡萄球菌和枝芽孢杆菌呈现交替演变，是虾酱发酵前期的优势产蛋白酶嗜盐菌。虾酱中的总可溶性蛋白随着发酵时间的延长逐渐下降。结论：葡萄球菌和枝芽孢杆菌是发酵前期虾酱中的优势产蛋白酶嗜盐菌，也是降解蛋白的主体微生物类群。

Objective： To isolate and identify protease- producing halophilic bacteria from the fermentation prophase of traditional Jinzhou shrimp paste. Methods： Protease-producing halophilic bacteria were isolated by 1.0% skimmed milk Gibbons test.These strains were identified by morphology, biochemical characteristics and molecular biology.Total soluble protein was analyzed using SDS-PAGE test.Results：16 strains protease-producing halophilic bacteria were obtained.Strain CW0- 1, CW0-2, CW0-3 and CW0-4 were identified as Vibro sp., strain CW1 -1, CW2-1 and CW2-2 were identified as Staphylococcus nepalensis, strain CW2-3, CWS-1, CW5-2 and CW5-3 were identified as Staphylococcus equorum, strain CW3-1 ,CW3-2, CW4-1 and CW4-2 were identified as Virgibacillus sp.,strain CW4-3 was identified as Virgibacillus dokdonensis.The Vibro sp.mainly existed in raw shrimp,halophilic Staphylococcus sp.were observed in fermentation second month and fifth month, Virgibacillus sp.were observed in fermentation third month to fourth month.Staphylococcus sp.and Virgibacillus sp.presented alternate successions, which were the dominant protease-producing halophilic bacteria in the fermentation prophase of shrimp paste. Total soluble protein was gradually decreased with the extension of fermentation time.Conclusion. Staphylococcus sp. And Virgibacillus sp.were the dominant protease-producing halophilic bacteria in the fermentation prophase of shrimp paste,and were considered as the dominant bacteria of degradation protein.