Department of Biochemistry, Faculty of Medicine, Srinakharinwirot University, Bangkok 10110, Thailand; E-Mails:
Int J Mol Sci. 2010 Sep 29;11(10):3783-92. doi: 10.3390/ijms11103783.
The aim of this study was to characterize the organic solvent and detergent tolerant properties of recombinant lipase isolated from thermotolerant Bacillus sp. RN2 (Lip-SBRN2). The isolation of the lipase-coding gene was achieved by the use of inverse and direct PCR. The complete DNA sequencing of the gene revealed that the lip-SBRN2 gene contains 576 nucleotides which corresponded to 192 deduced amino acids. The purified enzyme was homogeneous with the estimated molecular mass of 19 kDa as determined by SDS-PAGE and gel filtration. The Lip-SBRN2 was stable in a pH range of 9-11 and temperature range of 45-60 °C. The enzyme was a non metallo-monomeric protein and was active against pNP-caprylate (C8) and pNP-laurate (C12) and coconut oil. The Lip-SBRN2 exhibited a high level of activity in the presence of 108% benzene, 102.4% diethylether and 112% SDS. It is anticipated that the organic solvent and detergent tolerant enzyme secreted by Bacillus sp. RN2 will be applicable as catalysts for reaction in the presence of organic solvents and detergents.
本研究旨在研究从耐热芽孢杆菌 RN2（Lip-SBRN2）中分离得到的重组脂肪酶的有机溶剂和去污剂耐受性特性。采用反向和直接 PCR 法获得脂肪酶编码基因的分离。基因的完整 DNA 测序表明，Lip-SBRN2 基因包含 576 个核苷酸，对应于 192 个推导的氨基酸。通过 SDS-PAGE 和凝胶过滤测定，纯化的酶是均一的，估计分子量为 19 kDa。Lip-SBRN2 在 pH 值为 9-11 和温度范围为 45-60°C 的范围内稳定。该酶是非金属单体蛋白，对 pNP-辛酸酯（C8）和 pNP-月桂酸酯（C12）以及椰子油均具有活性。在存在 108%苯、102.4%二乙醚和 112%SDS 的情况下，Lip-SBRN2 表现出高活性。预计由芽孢杆菌 RN2 分泌的有机溶剂和去污剂耐受酶将适用于在有机溶剂和去污剂存在下的反应作为催化剂。
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