Identification of Protease-Producing Bacteria Isolated from Banyuwedang, Bali, and Characterization of its Protease

Q3 Environmental Science Squalen Bulletin of Marine and Fisheries Postharvest and Biotechnology Pub Date : 2018-12-30 DOI:10.15578/SQUALEN.V13I3.367

D. S. Zilda, Y. N. Fawzya, A. Uria

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引用次数: 4

Abstract

Proteases or peptidases is known as a largest group of hydrolytic enzymes and have been applied in various industries such as food, pharmacy, leather, detergent and waste treatment. Although they are also produced by plants and animals, microbes remain the main source of proteases in the world market which mostly derived from Bacillus sp. Aims of this research were to identify isolate BII-1 and study its protease. Analysis of 16Sr RNA sequencing showed the identity of BII-1 as Bacillus subtilis (99% similarity with the same species in GenBank). It was found that protease from BII-1 exhibited optimal temperature and pH of 50 oC and 8-9, respectively. It was activated by Li2+, Na2+, Mg2+ and K+. The degenerated primer for protease gene was designed, and a partial protease gene was amplified from BII-1. The sequencing result showed that this amplified gene shared 100 and 99% similarity with those from Geobacillus thermophiles and Bacillus subtilis in the GenBank, respectively.Keywords: protease, bacteria, Bacillus subtilis, Geobacillus thermophylus

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巴厘岛Banyuwedang产蛋白酶菌的鉴定及其蛋白酶的性质

蛋白酶或肽酶被称为最大的水解酶群，在食品、制药、皮革、洗涤剂和废物处理等各个行业都有应用。虽然它们也可以由植物和动物产生，但在世界市场上，微生物仍然是蛋白酶的主要来源，主要来源于芽孢杆菌。本研究的目的是鉴定分离物BII-1并对其蛋白酶进行研究。16Sr RNA测序结果显示，BII-1为枯草芽孢杆菌(Bacillus subtilis)，与GenBank同源菌株相似性达99%。结果表明，BII-1蛋白酶的最适温度和pH分别为50℃和8-9℃。它被Li2+、Na2+、Mg2+和K+激活。设计了蛋白酶基因的退化引物，并从BII-1中扩增出部分蛋白酶基因。测序结果表明，该扩增基因与GenBank中嗜热地杆菌和枯草芽孢杆菌基因的相似性分别为100%和99%。关键词:蛋白酶;细菌;枯草芽孢杆菌

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