Screening and characterization of a thermostable lipase from marine Streptomyces sp. strain W007
Screening and characterization of a thermostable lipase from marine Streptomyces sp. strain W007
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DOI：
10.1002/bab.1338
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发表时间：
2016-02
影响因子：
2.8
通讯作者：
Dongjuan Yuan;Dongming Lan;Ruipu Xin;Bo Yang;Yonghua Wang
中科院分区：
文献类型：
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作者：
Dongjuan Yuan;Dongming Lan;Ruipu Xin;Bo Yang;Yonghua Wang
采用沿着的方法，结合微生物基因组序列、两两比对和脂肪酶分类，寻找耐热脂肪酶。然后，利用毕赤酵母X-33表达了海洋链霉菌W 007的一种潜在的热稳定脂肪酶（命名为MAS 1），并对其生化性质进行了表征。脂肪酶MAS 1属于亚家族I.7，并且它与亚家族I.4中充分表征的枯草芽孢杆菌热稳定脂肪酶具有38%的同一性。纯化的酶估计为29 kDa。该酶的最适温度为40 °C，在60 °C下孵育1小时后仍保持80%以上的初始活性，表明MAS 1是一种热稳定的脂肪酶。MAS 1为碱性酶，最适pH为7.0，在pH 6.0 ~ 9.0范围内，酶活力稳定，在12 H内保持活力。在Cu ~（2+）、Ca ~（2+）、Ni ~（2+）和Mg ~（2+）存在下，该酶稳定性较好，保持了90%的初始酶活，而在乙二胺四乙酸存在下，该酶保持了89.05%的初始酶活。MAS 1对有机溶剂有一定的耐受性，但被多种表面活性剂抑制。MAS 1是一种甘油三酯脂肪酶，能水解甘油三酯和甘油二酯。这一结果为研究人员寻找具有工业应用价值的耐热脂肪酶提供了一个很好的范例。
A screening method along with the combination of genome sequence of microorganism, pairwise alignment, and lipase classification was used to search the thermostable lipase. Then, a potential thermostable lipase (named MAS1) from marine Streptomyces sp. strain W007 was expressed in Pichia pastoris X‐33, and the biochemical properties were characterized. Lipase MAS1 belongs to the subfamily I.7, and it has 38% identity to the well‐characterized Bacillus subtilis thermostable lipases in the subfamily I.4. The purified enzyme was estimated to be 29 kDa. The enzyme showed optimal temperature at 40 °C, and retained more than 80% of initial activity after 1 H incubation at 60 °C, suggesting that MAS1 was a thermostable lipase. MAS1 was an alkaline enzyme with optimal pH value at 7.0 and had stable activity for 12 H of incubation at pH 6.0–9.0. It was stable and retained about 90% of initial activity in the presence of Cu2+, Ca2+, Ni2+, and Mg2+, whereas 89.05% of the initial activity was retained when ethylene diamine tetraacetic acid was added. MAS1 showed the tolerance to organic solvents, but was inhibited by various surfactants. MAS1 was verified to be a triglyceride lipase and could hydrolyze triacylglycerol and diacylglycerol. The result represents a good example for researchers to discover thermostable lipase for industrial application.