Dukunde Amélie, Schneider Dominik, Lu Mingji, Brady Silja, Daniel Rolf
Department of Genomic and Applied Microbiology and Göttingen Genomics Laboratory, Institute of Microbiology and Genetics, Georg-August University Göttingen, Grisebachstr. 8, 37077, Göttingen, Germany.
Bayer AGET-TD-UP Biochemistry & Biocatalysis, Building B310, 207, 51368, Leverkusen, Germany.
Biotechnol Lett. 2017 Apr;39(4):577-587. doi: 10.1007/s10529-016-2282-1. Epub 2017 Jan 2.
To investigate the properties of a novel metagenome-derived member of the hormone-sensitive lipase family of lipolytic enzymes.
A forest soil metagenome-derived gene encoding an esterase (Est06) belonging to the hormone-sensitive lipase family of lipolytic enzymes was subcloned, heterologously expressed and characterized. Est06 is a polypeptide of 295 amino acids with a molecular mass of 31 kDa. The deduced protein sequence shares 61% similarity with a hypothetical protein from the marine symbiont Candidatus Entotheonella sp. TSY1. Purified Est06 exhibited high affinity for acyl esters with short-chain fatty acids, and showed optimum activity with p-nitrophenyl valerate (C5). Maximum enzymatic activity was at 50 °C and pH 7. Est06 exhibited high stability at moderate temperatures by retaining all of its catalytic activity below 30 °C over 13 days. Additionally, Est06 displayed high stability between pH 5 and 9. Esterase activity was not inhibited by metal ions or detergents, although organic solvents decreased activity.
The combination of Est06 properties place it among novel biocatalysts that have potential for industrial use including low temperature applications.
研究一种源自宏基因组的新型激素敏感脂肪酶家族脂解酶的特性。
对一个编码属于激素敏感脂肪酶家族脂解酶的酯酶（Est06）的森林土壤宏基因组来源基因进行了亚克隆、异源表达及特性鉴定。Est06是一个由295个氨基酸组成的多肽，分子量为31 kDa。推导的蛋白质序列与海洋共生菌暂定种Entotheonella sp. TSY1的一种假定蛋白质具有61%的相似性。纯化后的Est06对含短链脂肪酸的酰基酯表现出高亲和力，对戊酸对硝基苯酯（C5）表现出最佳活性。最大酶活性出现在50℃和pH 7条件下。Est06在中等温度下表现出高稳定性，在13天内30℃以下能保持其所有催化活性。此外，Est06在pH 5至9之间表现出高稳定性。酯酶活性不受金属离子或去污剂抑制，不过有机溶剂会降低其活性。
Est06的特性使其成为具有工业应用潜力（包括低温应用）的新型生物催化剂之一。
