School of Life Science and Biotechnology, Dalian University of Technology, No. 2 Linggong Road, Ganjingzi District, Dalian, 116024, China.
School of Food and Environmental Science and Technology, Dalian University of Technology, Panjin, 124221, China.
Extremophiles. 2018 Mar;22(2):287-300. doi: 10.1007/s00792-018-0997-8. Epub 2018 Jan 13.
By screening 25 different psychrophilic strains isolated from the Arctic habitat, we isolated a strain capable of producing lipase. We identified this strain as Psychrobacter sp. ZY124 based on the amplified 16S rDNA sequence. The lipase, named as Lipase ZC12, produced from the supernatant of Psychrobacter sp. ZY124 cultured at 15 °C was purified to homogeneity by ammonium sulfate precipitation followed by Phenyl Sepharose FF gel hydrophobic chromatography. Based on the obtained amino acid sequence, Lipase ZC12 is classified as a member of the Proteus/psychrophilic subfamily of lipase family I.1; it has a molecular weight of 37.9 kDa. We also determined that the apparent optimum temperature for Lipase ZC12 activity is 40 °C. Lipase ZC12 shows remarkable organic solvent tolerance by remaining more 50% after incubated with 10-90% different organic solvents. In addition, acyl chain esters with C12 or longer were confirmed to be preferable substrates for Lipase ZC12. Lipase ZC12 also shows better stereoselectivity for (R, S)-1-phenylethanol chiral resolution in n-hexane solvent with (S)-1-phenylethanol (ee 92%) and conversion rate (39%) by transesterification reactions. These properties may provide potential applications in biocatalysis and biotransformation in non-aqueous media, such as in detergent, transesterification or esterification and chiral resolution.
通过筛选来自北极栖息地的 25 种不同的嗜冷菌株，我们分离出了一株能够产生脂肪酶的菌株。我们根据扩增的 16S rDNA 序列将该菌株鉴定为 Psychrobacter sp. ZY124。脂肪酶命名为 Lipase ZC12，由在 15°C 下培养的 Psychrobacter sp. ZY124 的上清液产生，通过硫酸铵沉淀和苯基 Sepharose FF 凝胶疏水层析纯化至均一性。根据获得的氨基酸序列，Lipase ZC12 被归类为脂肪酶家族 I.1 的 Proteus/嗜冷亚家族的成员；它的分子量为 37.9 kDa。我们还确定了 Lipase ZC12 的表观最适温度为 40°C。Lipase ZC12 具有显著的有机溶剂耐受性，在与 10-90%的不同有机溶剂孵育后，仍保持超过 50%的活性。此外，C12 或更长的酰基链酯被证实是 Lipase ZC12 的优选底物。Lipase ZC12 在正己烷溶剂中通过酯交换反应对 (R, S)-1-苯乙醇手性拆分也表现出更好的立体选择性，得到 (S)-1-苯乙醇（ee 值为 92%）和转化率（39%）。这些特性可能为非水介质中的生物催化和生物转化提供潜在应用，例如在洗涤剂、酯交换或酯化以及手性拆分中。
