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从马拉巴里山羊体内分离出的新型瘤胃细菌——假单胞菌属BUP6菌株脂肪酶的生产、优化及部分纯化

Production, optimization, and partial purification of lipase from Pseudomonas sp. strain BUP6, a novel rumen bacterium characterized from Malabari goat.

作者信息

Priji Prakasan, Unni Kizhakkepowathial Nair, Sajith Sreedharan, Binod Parameswaran, Benjamin Sailas

机构信息

Enzyme Technology Laboratory, Biotechnology Division, Department of Botany, University of Calicut, Thenhipalam, Kerala, India.

出版信息

Biotechnol Appl Biochem. 2015 Jan-Feb;62(1):71-8. doi: 10.1002/bab.1237. Epub 2014 Jul 23.

Abstract

This study introduces a novel bacterium-Pseudomonas sp. strain BUP6-isolated from the rumen of the Malabari goat with efficiency for producing lipase. It showed significant production of lipase when grown in a newly designed basal medium, supplemented with vegetable oil. Suitability of five vegetable oils such as groundnut oil, coconut oil, olive oil, sunflower oil, and palm oil as inducer for the production of lipase was examined, and groundnut oil supported the highest production of lipase (96.15 U/mL). Various physical parameters required for the maximum production of lipase were statistically optimized. Plackett-Burmann design was employed to study the interactive effects of physical parameters and found that temperature, agitation, and pH effected the production of lipase significantly. The optimum conditions for lipase production (37 °C, 200 rpm, and pH 6.9) were detected by Box-Behnken design and response surface methodology, which resulted in the 0.3-fold increase (i.e., 126 U/mL) of the lipase activity over the unoptimized condition. The apparent molecular mass of partially purified lipase was 35 kDa, as judged by SDS-PAGE; the activity of lipase was also confirmed by native PAGE. Thus, this study focuses on the need for the exploitation of rumen microbes for the production of industrially significant and human-friendly biomolecules to meet the future needs.

摘要

本研究介绍了一种从马拉巴山羊瘤胃中分离出的新型细菌——假单胞菌属菌株BUP6,它具有高效生产脂肪酶的能力。当在新设计的添加植物油的基础培养基中生长时,它表现出显著的脂肪酶产量。研究了花生油、椰子油、橄榄油、向日葵油和棕榈油这五种植物油作为脂肪酶生产诱导剂的适用性,其中花生油支持最高的脂肪酶产量(96.15 U/mL)。对脂肪酶最大产量所需的各种物理参数进行了统计优化。采用Plackett-Burmann设计研究物理参数的交互作用,发现温度、搅拌和pH对脂肪酶的产量有显著影响。通过Box-Behnken设计和响应面方法检测到脂肪酶生产的最佳条件(37°C、200 rpm和pH 6.9),与未优化条件相比,脂肪酶活性提高了0.3倍(即126 U/mL)。通过SDS-PAGE判断,部分纯化的脂肪酶的表观分子量为35 kDa;天然PAGE也证实了脂肪酶的活性。因此,本研究强调了利用瘤胃微生物生产具有工业重要性且对人类友好的生物分子以满足未来需求的必要性。

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