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Biotechnol Rep (Amst). 2018 Dec 4;21:e00295. doi: 10.1016/j.btre.2018.e00295. eCollection 2019 Mar.

Cold-active extracellular lipase: Expression in Sf9 insect cells, purification, and catalysis.

Author information

1
Molecular Endocrinology and Nephrology, Axe CHU Research Center and Department of Molecular Medicine, Laval University, 2705 boulevard Laurier, Québec, G1V 4G2, Canada.
2
Laboratory of Sustainable Development of Polar Fishery, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, 106 Nanjing Road, Qingdao 266071, China.

Abstract

Cold-active lipases are gaining special attention nowadays as they are increasingly used in various industries such as fine chemical synthesis, food processing, and washer detergent. In the present study, an extracellular lipase gene from Yarrowia lipolytica (LIPY8) was cloned and expressed by baculovirus expression system. The recombinant lipase (LipY8p) was purified using chromatographic techniques, resulting in a purification factor of 25.7-fold with a specific activity of 1102.9U/mg toward olive oil. The apparent molecular mass of purified LipY8p was 40 kDa. The enzyme was most active at pH 7.5 and 17 °C. It exhibited maximum activity toward medium chain (C10) esters. The presence of transition metals such as Zn2+, Cu2+, and Ni2+ strongly inhibited the enzyme activity, which was enhanced by EDTA. The lipase activity was affected by detergents and was elevated by various organic solvents at 10% (v/v). These enzymatic properties make this lipase of considerable potential for biotechnological applications.

KEYWORDS:

Baculovirus expression system; C12E8, octaethylene glycol monododecyl ether; Cold-active; DMF, Dimethylformamide; Extracellular lipase; PH, polyhedrin; Purification; RhB, rhodamine B; RhB-OOe, RhB-olive oil; Yarrowia lipolytica; pNPA, p-nitro phenyl acetate; pNPB, p-nitro phenyl butyrate; pNPD, p-nitro phenyl decanoate; pNPL, p-nitro phenyl dodecanoate; pNPM, p-nitro phenyl myristate; pNPP, p-nitro phenyl palmitate; β-DDM, n-Dodecyl-β-d-Maltoside; β-ME, β-mercaptoethanol; β-OG, n-octyl-β-d-glucoside

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