Lipase Production From Fusarium Sp. SYBC Lz And The Enzyme Characterization

Lipase, also known as Glycerides hydrolase, which is a kind of effective hydrolase thatcatalyze water-fast lipid for instance glyceride, has been applied abundantly in food, medicine,environment, weave, organic synthesis and so and. However, its utilization is limited by highproduction cost, it is unable quitely to meet a great quantity of the industrial requirement.Therefore, investigating lipase has still significance.The paper that included identification of bacterium, optimization of fermentation medium,influence of additives, purification of lipase and the basic characterization of enzyme hadarrived at several conclusions in follows:（1）Accroding to the D1/D2of28S rRNA gene sequence, the investigative strain of theessay was uniform branch and Max Identities was99%contrasted with Fusariumproliferatum, so it was belong to Fusarium and termed temporarily as Fusarium sp. SYBCLz.（2）The fermentation conditions of Fusarium sp. SYBC Lz was investigated. The optimizedmedium contained: the glucose12.0g/L, the peptone8.0g/L, NaH2PO43g/L, mutton fat8.0g/L and emulsifier OP50mL/L. The lipase activity of optimization attained4.51U/mL and itwas enhanced3.46folds of that before optimization.（3）It had an effect on lipase production of Fusarium sp. SYBC Lz by adding Gly, Lys, Asp,acetic acid, propanoic acid and soapberry extraction to fermentation medium, in which theaffect of the soapberry extraction was marked.（4）Then, electrophoretically pure lipase （LIP Z） was obtained by freeze-drying, theprecipitation of （NH4）2SO4, Aqueous Two-phase System, DEAE-cellulose ion-exchangechromatography and G-75gel filtration chromatography. LPI Z was ultimatily purified13.89folds.（5）At last, the characteristics of LIP Z was also investigated,which pointed out its optimaltemperature and pH was25℃and9.5respectively,which proved that it was a cold-adaptedalkaline lipase;the best substrates of the lipase was p-NPB; the lipase was stimulatedconsiderably by K⁺, Fe³⁺and Cu²⁺while inhibited by EDTA, Zn²⁺and Mg²⁺; moreover, thelipase was resistant to alcohol; Vmax=72.4U/mg, Km=0.67mmol/L; its N-terminal amion acidsequence was GEVSQDLFNQ which revealed it was a novel Fusarium lipase.