Purification and characterization of hepatic triacylglycerol lipase isolated from rainbow trout,Oncorhynchus mykiss

Trialcylglycerol (TG) lipase was isolated and partially purified from rainbow trout liver. Triacylglycerol lipase activity was assayed by measuring¹⁴C-oleic acid release from¹⁴C-triolein.¹⁴C-oleic acid release was linear for up to two hours. Optimal activity occurred at pH 7.0 and 15°C. Most of the lipase activity was recovered in the cytosolic fraction. A 27,000-fold purification was achieved after Sepharose (Bio-gel A 0.5 M, 200–400 mesh) chromatography of a resuspended 20% ammonium sulfate fraction. The molecular weight of the trout hepatic lipase as determined by size-exclusion chromatography and by SDS-polyacrylamide gel electrophoresis was 40–43 kD. Lipase-mediated hydrolysis of TG resulted in the production of diacylglycerols, monoacylglycerols, and fatty acids. Kinetic analysis indicated that V_max=0.016 nmol/h/mg protein and that K_m=0.28 mM triolein. Lipolytic activity was enhanced in the presence of cAMP/ATP-Mg²⁺. These results suggest that the liver of trout possesses a neutral TG lipase that is responsible for mobilizing stored TG and is catalytically activated by phosphorylation.A part of this work was presented at the Annual Meeting of the American Society of Zoologists, December 26–30, 1990, San Antonio, TX.     

卵黄和低密度脂蛋白对冷冻保存绵羊精子胆固醇、磷脂含量的影响

为了探索绵羊冷冻精液受精率低的原因,在稀释液中用低密度脂蛋白代替卵黄冷冻绵羊精液,研究LDL对冷冻中精子脂类含量的影响。结果表明,以卵黄稀释液冷冻精液,精子胆固醇含量从鲜精的234．54nmol／10^9降低到163．76nmol／10^9精子,胆固醇与磷脂的比值（c／p）从0．54下降到0．36;以9g／100mL LDL代替卵黄,冷冻后胆固醇含量为243．73nmol／10^9精子,c／p值保持在0．50。研究得出在卵黄稀释液中冷冻绵羊精液,精子胆固醇流失,c／p值下降;以LDL代替卵黄,可有效保护精子胆固醇,避免流失,维持了精子c／p值。     

家蚕对BmNPV抗病性与中肠消化酶活性的关系

对6个现行家蚕品种中肠消化酶活性与家蚕对BmNPV病毒抗性比较与分析,以期寻找现行家蚕品种消化酶的活性与抗性的关系,为辅助家蚕育种及品种推广奠定基础.以DNS法测定不同品种海藻糖酶活性,以Fo-lion-酚法测定不同品种蛋白酶活性.以及以铜皂法测定脂肪酶活性,同时对不同品种家蚕4龄起蚕经口添加BmN-PV病毒进行抗性调查分析.结果表明,蛋白酶、脂肪酶酶活顺序为664×663较低,667×668相对较高,其他品种处于中间层次,对应抗性测定结果的相关分析表明,活性越强抗性越强,呈显著正相关关系;而海藻糖酶的活性与抗性的关系没有较好的相关性.因此,家蚕中肠消化液的蛋白酶、脂肪酶可能与家蚕对BmNPV病毒抗性有关.     

罗非鱼鱼皮提取明胶的酶法脱脂工艺研究

用脂肪酶对罗非鱼鱼皮进行酶法脱脂工艺研究。通过单因素与正交试验确定最适的酶法脱脂工艺为:pH值9,脱脂时间3h,酶添加量2%,料液比1:4,温度40℃,该工艺的脱脂率可达67.5%。通过超声波辅助脱脂研究表明其脱脂率并无明显的提高。     

奥尼罗非鱼淀粉酶、脂肪酶的分布与特性

以奥尼罗非鱼(Oreochromisniloticus×O.aureus)为实验材料,研究其淀粉酶、脂肪酶分布与特性。结果表明,Ⅰ(体重55.14g)、Ⅱ(体重122.82g)、Ⅲ(体重225.68g)组实验鱼肠道淀粉酶、脂肪酶活性均表现出相似规律:活性从大到小依次为前肠、中肠、后肠。实验鱼体重从55g增至122g,肠道淀粉酶活性急剧上升,实验鱼体重从122g增至225g,酶活性增幅减缓。3组实验鱼肠道各肠段淀粉酶活性相对比值(前肠、中肠、后肠)分别为1:0.95:0.52、1:0.44:0.21和1:0.50:0.24;脂肪酶活性相对比值(前肠、中肠、后肠)分别为1:0.17:0.12、1:0.34:0.14和1:0.98:0.64。3组实验鱼淀粉酶活性相对比值(Ⅰ、Ⅱ、Ⅲ组)在全肠、肝胰脏中分别为1:8.25:18.94和1:3.43:9.31;3组实验鱼脂肪酶活性相对比值(Ⅰ、Ⅱ、Ⅲ组)在全肠中为1:3.82:13.09。奥尼罗非鱼肠道、肝胰脏淀粉酶最适pH值分别为6.5～7 0和7.5;最适反应温度均为30℃;最适反应底物浓度均为0.8%。肠道脂肪酶最适pH值为7.0～7.5,最适反应温度为35℃。     

添加外源脂肪酶对瓦氏黄颡鱼的生长、消化酶及血清生化指标的影响

为考察添加脂肪酶酶制剂对瓦氏黄颡鱼生长、消化酶以及血清生化指标的影响,在基础饲料中分别添加0mg/kg、100mg/kg、300mg/kg、500mg/kg脂肪酶制剂,饲喂平均体重(3.47±0.05)g的瓦氏黄颡鱼90d。试验结果表明:添加脂肪酶100mg/kg、300mg/kg、500mg/kg组分别比对照组增重率提高3.86%(P>0.05)、9.29%(P<0.05)、6.75%(P>0.05),饲料系数降低了3.17%(P>0.05)、6.35%(P<0.05)、5.29%(P<0.05)。同时添加脂肪酶各组的血清甘油三酯、低密度脂蛋白和总胆固醇含量都显著低于对照组(P<0.05),通过消化酶测定表明,脂肪酶添加各组均显著提高了胰蛋白酶和肠淀粉酶活性(P<0.05),100mg/kg脂肪酶组还显著提高了胰脂肪酶和胰淀粉酶(P<0.05),300mg/kg脂肪酶组胰脂肪酶活性最高(P<0.05)。上述研究表明,脂肪酶在瓦氏黄颡鱼饲料中的适当添加量为300mg/kg为宜。     

山楂黄酮提取物降血脂研究

[目的]研究山楂黄酮提取物的降血脂作用,为其相关产品开发应用提供依据。[方法]采用渗漉法提取山楂黄酮,对SD大鼠进行试验,待大鼠形成高脂模型后,采用低、中、高剂量灌胃山楂黄酮提取物,30 d后测定大鼠血脂中总胆固醇、甘油三酯、低密度脂蛋白、高密度脂蛋白的变化。[结果]山楂黄酮提取物使各剂量组大鼠血脂中的总胆固醇、低密度脂蛋白含量明显降低,对甘油三脂、高密度脂蛋白有调节作用。[结论]山楂黄酮提取物具有显著的降血脂功效。     

Use of tween 20 as a substrate for assay of lipase activity in soils

Abstract

A method for assaying the soil lipase activity is described. It involves the titrimetric estimation of the amount of lauric acid released by the lipase activity when the soil is incubated with Tween 20 in the presence of toluene at 30°C for 18 h under agitation. The method is simple and precise and incubation without agitation is also possible. The method has been applied to six different kinds of soils. The lipase activity in the cultivated soils ranged from 22.5 to 75.5 mmol min^?1 g^?1 of dried soil. The K _m value for Tween 20 was 1.8 × 10^?4 m. The optimum pH was approximately 7.5. The hydrolysis of liveen 20 in soil was inhibited by glycerol which was the essential moiety of glyceride. The inhibition by glycerol was found to be competitive. These results indicate that Tween 20 is a potential substrate for the assay of the glyceride hydrolytic activity in soils.     

日粮能量水平对扬州鹅脂肪代谢及HL基因表达的影响

选用35日龄、体重相近的扬州鹅200只,随机分成4组,每组5个重复,每个重复10只(公、母各半),分别饲喂4个代谢能水平的日粮(A组10.11、B组11.19、C组12.37和D组13.45 MJ/kg),研究其对扬州鹅脂肪代谢及肝脂酶(HL)基因表达的影响。结果表明:与A组相比,D组扬州鹅的屠宰率显著提高(P<0.05),腹脂率极显著提高(P<0.01);随着能量水平的提高,血清极低密度脂蛋白和甘油三酯显著降低,低密度脂蛋白、高密度脂蛋白、脂蛋白脂酶、肝脂酶活性及其表达量均显著增加(P<0.05)。本研究结果表明,日粮能量水平显著影响扬州鹅机体脂肪代谢和HL基因mRNA的表达。     

脂肪酶法催化胆固醇合成胆固醇棕榈酸酯的研究

探讨了固定化Candida antarctica脂肪酶催化胆固醇与棕榈酸合成胆固醇棕榈酸酯的反应过程。通过脂肪酶催化酯化反应的工艺路线进行研究,系统考察了转速、反应介质、醇酸比、固定化酶用量、反应温度等对酯化反应的影响。结果表明,以环己烷为溶剂,在30℃,胆固醇与棕榈酸物质的量比1：1,2%（质量分数）脂肪酶,反应时间为48h的条件下,反应的最高酯化率可达83%以上,产品经HPLC和LC-MS分析的纯度可达98%以上;固定化C.antarctica脂肪酶具有较高的催化稳定性,其半衰期至少达120d。