Improvement of extracellular lipase production by a newly isolated Yarrowia lipolytica mutant and its application in the biosynthesis of L-ascorbyl palmitate

Abstract

Yarrowia lipolytica Wt-11 producing an extracellular lipase was isolated and identified. To improve the lipase production, Y. lipolytica Wt-11 was subjected to low-energy ion implantation mutation breeding, and a best mutant, Y. lipolytica Mut-96, was obtained after screening. Under the optimal cultivation conditions, the scaled-up production of lipases were performed, and the lipase activity of Y. lipolytica Mut-96 was enhanced nearly 5.5-fold compared with that of Y. lipolytica Wt-11. After fermentation, the lipases were purified, and the characteristics of the purified lipases were studied. The optimum temperatures and pHs for lipases from Wt-11and Mut-96 were 30°C and 8.0, respectively. The purified lipases were stable between pH 7.0 and 8.5 and unstable at temperatures above 40°C. The lipase activities were enhanced by Ca²⁺, Ba²⁺, Mn²⁺, Fe²⁺ and SDS. The synthesis of L-ascorbyl palmitate via esterification with L-ascorbic acid and palmitic acid by immobilized lipases from Wt-11 and Mut-96 in organic media was investigated, and the L-ascorbyl palmitate can be respectively produced at levels of 14.8 and 27.5g/L.

Keywords: Breeding; L-ascorbyl palmitate; Lipase; Low-energy ion implantation; Yarrowia lipolytica.