EFFECTS OF DIFFERENT PARAMETERS ON THE LIPOLYTIC ACTIVITIES OF LIPASE AND PHOSPHOLIPASE EXTRACTED FROM PRUNUS ARMINICA MEAL OF HUNZA CULTIVAR

A. Hifza
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Abstract

Prunus Arminica.L seeds of Hunza cultivar were defatted with chloroform, methanol mixture (2:1v/v) by ultrasonic assisted extraction technique. The lipase and phospholipase extracted, incubated at 400C and pH 6 in citrate buffer for 1 hour. The lipolytic activities of enzymes were measured at different temperatures i,e 30-700C , pH i,e 3-9 and solvents i,e n-heptane , Di-isopropyl ether and Cyclo hexane. 10 % emulsion of lecithin and olive oil used as the substrate for phospholipase and lipase substrates respectively. The enzymes showed stability at lower temperatures i,e 35-500C and slightly acidic to neutral i.e pH 5 to pH 7. Concentration of liberated fatty acids was the indicator of the enzymes activity which were measured by taking absorbance at 440nm. Stearic acid with different concentrations in hexane and methanol (1:1v/v) were used for the standard curve. Cu-TEA reagent with small quantity of CaCl2 was used for maximum extraction of Cu-FFA and for stable value of blanks in the present study. The enzymes showed maximum activities at 450 C temperature and 6 pH.
不同参数对hunza品种野李粕中脂肪酶和磷脂酶溶脂活性的影响
李属Arminica。以氯仿与甲醇(2:1v/v)的混合物为溶剂,采用超声辅助提取技术对罕萨L种子进行脱脂。提取脂肪酶和磷脂酶,在400C和pH 6柠檬酸缓冲液中孵育1小时。测定了酶在不同温度(30 ~ 700c)、pH(1)、pH(3 ~ 9)和溶剂(1)、正庚烷、二异丙醚和环己烷)下的溶脂活性。10%卵磷脂乳液和橄榄油分别作为磷脂酶底物和脂肪酶底物。酶在较低温度(35 ~ 500℃)和微酸性至中性(pH 5 ~ pH 7)下表现出稳定性。用440nm吸光度法测定酶活性,并以游离脂肪酸浓度为指标。采用硬脂酸在正己烷和甲醇中不同浓度(1:1v/v)作标准曲线。本研究采用少量CaCl2的Cu-TEA试剂最大限度地提取Cu-FFA,并使空白值稳定。酶在450℃温度和6 pH条件下活性最高。
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