Molecular cloning and characterization of a novel cold-active lipase from Pichia lynferdii NRRL Y-7723

Authors: BAE, JAE-HAN - Kyungpook National University; KIM, IN-HWAN - Korea University; LEE, KI-TEAK - Chungnam National University; Hou, Ching; KIM, HAK-RYUL - Kyungpook National University

Submitted to: Biocatalysis and Agricultural Biotechnology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 5/26/2017
Publication Date: 7/1/2017
Citation: Bae, J.H., Kim, I.H., Lee, K.T., Hou, C.T., Kim, H.R. 2017. Molecular cloning and characterization of a novel cold-active lipase from Pichia lynferdii NRRL Y-7723. Biocatalysis and Agricultural Biotechnology. 11:19-25. doi: 10.1016/j.bcab.2017.05.008.
DOI: https://doi.org/10.1016/j.bcab.2017.05.008

Interpretive Summary: Lipase is an important enzyme used in a broad range of industrial applications for degradation of fats. For application in restricted reaction conditions, lipases often require special characteristics such as activity with low temperatures. In our previous study, we screened several yeasts for the production of extracellular cold-active lipase and reported the identification of a novel cold-active lipase produced from Pichia lynferdii NRRL Y-7723 [J. Agric. Food Chem. 58:1322-1326. 2010]. In addition, we achieved remarkable increase of lipase production from this strain over 10 times compared to that in normal conditions by optimization study [J. Agric. Food Chem. 61:882-886. 2013]. We purified and characterized the cold-active lipase from this strain [Biotechnology and Bioprocess Engineering 19:851-859. 2014]. In this study, we report the identification of lip1 gene that encodes an extracellular cold-active lipase from this strain. The lip1 gene was cloned into a bacterial expression vector, and the recombinant lipase was successfully expressed and purified. Several parameters of the recombinant lipase were analyzed, and Lip1 showed high activity at low temperature. The results of this study may also be used in the preparation of functional/health foods and improve the health and nutrition of humankind.

Technical Abstract: Lipase is one of the widely used biocatalysts, which is well studied for its application in industrial production. Recently, lipases with special characteristics such as thermo-stability, alkaline-, acidic nature, and cold-activity, have gained attention for effective applications in specific purposes. Previously, Pichia lynferdii NRRL Y-7723 was reported to produce high amounts of extracellular cold-active lipase (J Agric Food Chem 2010, 58:1322–1326). In this study, we report the identification of Lip1 gene that encodes an extracellular cold-active lipase from P. lynferdii NRRL Y-7723. The open reading frame of the gene consisted of 1122 bp of nucleotides that encoded a protein with 373 amino acids. The deduced molecular weight and isoelectric point were 41.8 kDa and pH 5.82, respectively. The Lip1 gene was cloned into a bacterial expression vector, and the recombinant lipase was successfully expressed and purified. Several parameters of the recombinant lipase were analyzed, and Lip1 showed high activity at low temperature.