Intraspecific diversity of Aureobasidium pullulans strains from different marine environments

Totally more than 500 yeast strains were isolated from seawater, sea sediments, mud of sea salterns, marine fish guts and marine algae. The results of routine and molecular biology identification methods show that nine strains among these marine yeasts belong to Aureobasidium pullulans, although the morphologies of their colonies are very different. The marine yeasts isolated from different marine environments indicate that A. pullulans is widely distributed in different environmental conditions. These Aureobasidium pullulans strains include A. pullulans 4#2, A. pullulans N13d, A. pullulans HN3-11, A. pullulans HN2-3, A. pullulans JHSc, A. pullulans HN4.7, A. pullulans HN5.3, A. pullulans HN6.2 and A. pullulans W13a. A. pullulans 4#2 could produce cellulase and single cell protein. A. pullulans N13d could produce protease, lipase, amylase and cellulase. Both A. pullulans HN3-11 and A. pullulans HN2-3 were able to produce protease, lipase and cellulase. A. pullulans JHSc could secrete cellulase and killer toxin. Both A. pullulans HN4.7 and A. pullulans HN5.3 could yield lipase and cellulase. A. pullulans W13a was able to secrete extracellular amylase and cellulase while A. pullulans HN4.7 and A. pullulans N13d could produce siderophores. This means that different A. pullulans strains from different marine environments have different physiological characteristics, which may be applied in many different biotechnological industries.     

Adsorption of extracellular proteases in seawater onto filters during size fractionation

In aquatic environments extracellular enzymes are bound to microbial cells or exist in a free and adsorbed state. Various filters have been used to fractionate these enzymatic activities, but enzymes may be readily adsorbed onto some materials, and such adsorption can induce errors in the estimation of enzymatic activity. In this study we examined three filters to determine the most suitable filter for fractionation when estimating proteolytic enzyme activity in seawater. We found that the polycarbonate Nuclepore membrane, widely used for size fractionation because of its pore-size accuracy, was the most favorable for this purpose, even though it adsorbed slightly more enzymes than the low-protein-binding polyethersulfone membrane. We also found that trypsin-and chymotrypsin-type enzymes were more easily adsorbed than aminopeptidases.     

对虾蛋白自溶制备ACE抑制肽的工艺条件优化

以凡纳滨对虾为原料，以ACE抑制率为指标，利用响应面法对虾肉蛋白自溶制备ACE抑制肽的工艺条件进行了优化，即在酶解条件（pH值、温度、虾头虾肉质量比）和ACE（Angiotensin I-converting Enzyme，ACE）抑制率之间建立了数学模型Y=23．59—0．21X1＋0．84X2＋0．85X3—0．71X1^2-0．94X2^2-1．06X3^2＋0．088X1X2—0．46X1X3-0．87X2X3。分析表明，在3个因素中，虾头与虾肉比例对ACE抑制率的影响最为显著。优化后的工艺参数为：pH7．35，温度57．2℃，虾头与虾肉比例为1：1。根据回归方程的预测结果，反应时间为4h，其ACE抑制率达41．9％。     

Effects of temperature, pH and NaCl on protease activity in digestive tract of young turbot, Scophthalmus maximus

1 INTRODUCTION Digestive enzyme activity is one of important issues for learning digestive physiology of fish, and widely applied in commercial fish culture. So far, the data obtained in fish showed that the diges- tive enzymes were qualitatively similar …     

Screening of protease-producing marine yeasts for production of the bioactive peptides

Over 400 yeast strains from seawater and sediments were obtained, but only five strains named HN2 -3, N13d, N13C, Mb5 and HN3 - 2 among them could form clear zones around their colonies on the double plates with 2.0% casein. Peptides in the hydrolysate produced by the proteases from strains HN2 -3 and N13d had higher angiotensin I-converting-enzyme （ACE）-inhibitory activity. The two marine yeast strains were identified to be Aureobasidium pullulans according to the results of routine yeast identification and molecular methods. After purification of the proteases from the two marine yeast strains, it was found that the optimal pH for them was both 9.0, both of them were serine alkaline protease. However, the optimal temperature for the protease from the strain HN2 -3 was 52℃ while that from strain N13d was 48℃. ACE-inhibitory activity of the peptides in the hydrolysate of shrimp protein produced by the purified protease from the strain HN2 -3 was the highest while antioxidant activity in the hydrolysate of spirulina protein produced by the purified protease from the strain N13d was the highest.     

饲料中大豆异黄酮和大豆皂甙对牙鲆肝脏和肠道蛋白质消化酶活性和基因表达的影响

以鱼粉为蛋白源,小麦粉为糖源,鱼油为脂肪源配制基础饲料,分别添加不同浓度梯度(0.1%、0.4%和0.8%)的大豆异黄酮和大豆皂甙。以不添加大豆异黄酮和大豆皂甙的基础饲料作为对照组,共制成7种等氮(粗蛋白49.1%)、等能(总能20.1kJ/g)的实验饲料。在循环水系统中养殖初始体质量为(2.58±0.01)g的牙鲆(Paralichthys olivaceus)幼鱼,8周后观测牙鲆肝脏和肠道蛋白质消化酶活性和基因表达的影响。结果表明,牙鲆肠道胰蛋白酶的活性不受饲料中大豆异黄酮的显著影响(P>0.05),却随大豆皂甙添加量的升高而显著下降(P<0.05)。肠道糜蛋白酶的活性在饲料中大豆异黄酮添加量为0.1%时取得最大值(37.3±5.9)U/mg,显著高于其它处理组。饲料中添加不同水平的大豆异黄酮对肝脏中胰蛋白酶-1和糜蛋白酶-1基因表达量有显著(P<0.05)的影响,而添加大豆皂甙却对这些基因表达量的影响不显著。由此可见,大豆异黄酮直接作用于基因表达和酶活两个水平,而大豆皂甙只在酶活水平发挥显著的作用。同时,大豆异黄酮对牙鲆饲料利用率影响的双重性依赖于剂量的大小,其中的作用机制有待于进一步研究。     

对虾蛋白自溶制备ACE抑制肽的工艺条件优化

以凡纳滨对虾为原料,以ACE抑制率为指标,利用响应面法对虾肉蛋白自溶制备ACE抑制肽的工艺条件进行了优化,即在酶解条件(pH值、温度、虾头虾肉质量比)和ACE(Angiotensin I-converting Enzyme,ACE)抑制率之间建立了数学模型Y=23.59-0.21X1+0.84X2+0.85X3-0.71X12-0.94X22-1.06X32+0.088X1X2-0.46X1X3-0.87X2X3。分析表明,在3个因素中,虾头与虾肉比例对ACE抑制率的影响最为显著。优化后的工艺参数为:pH7.35,温度57.2℃,虾头与虾肉比例为1∶1。根据回归方程的预测结果,反应时间为4 h,其ACE抑制率达41.9%。     

温度对不同规格欧鳗消化器官蛋白酶和淀粉酶活性影响的初步研究

本试验设置的8个反应温度组为5℃，15℃，20℃，25℃，30℃，35℃，45℃和55℃，欧鳗分51．5～57．5g，92．5～99．5g，128．5～136g，226～233g4种规格，试验结果表明，蛋白酶活性在4种规格均表现为肠＞胃＞肝，并均随温度升高而增强。在45℃蛋白酶活性最强，其中51．5～57．5g规格组肠蛋白酶活性显著较高．淀粉酶活性，基本上随着温度升高活力升高，温度在25℃时活性最高，以后其活性又开始下降。肝、肠、胃淀粉酶活性在两种较小规格下，胃＞肠＞肝，在较大规格，肝＞肠＞胃。     

海洋弧菌碱性蛋白酶的分离纯化及部分性质研究

采用硫酸铵沉淀、Sephadex- 75 ,Sephadex- 10 0凝胶过滤层析等方法纯化海洋弧菌 (Vibriop acini) X4 B- 7菌株产生的碱性蛋白酶 ,得到电泳纯酶制品 ,并对纯化酶的性质进行了研究。结果显示 :纯酶的分子量为 2 7KD,等电点 p I=8.7,最适反应 p H9.0～ 10 .5 ,最适反应温度 5 0～ 6 0℃。ED-TA对酶活力没有影响 ,高酶浓度可以降低 SDS对酶的抑制作用 ,该酶可用于解聚组蛋白。 DNA琼脂糖凝胶电泳证明 :酶对 DNA酶有降解作用 ,而对 DNA没有降解作用 ,该酶有希望应用于核酸的提取     

人工养殖长鳍篮子鱼消化道指数及3 种消化酶活性分布

以体质量58.51 g±21.15 g、体长13.08 cm±1.856 cm的长鳍篮子鱼(Siganus canaliculatus)为实验材料,对其消化道指数和主要消化酶活性分布进行了研究.采用常规方法测定了长鳍篮子鱼的消化道指数,其比内脏重、比肝重、比胃重,比幽门盲囊重、比肠重、比肠长分别为0.1572±0.0230、0.0166±0.0060、0.0115±0.0070、0.004 3±0.002 1、0.020 2±0.0102、2.595±0.457;体质量与体长的回归方程为Y=0.080 6X2.5457(r=0.9777,P<0.01).蛋白酶在各消化器官中的比活力顺序为幽门盲囊>肠>肝脏>胃;淀粉酶的比活力顺序为肠>幽门盲囊>胃>肝脏;脂肪酶比活力顺序为幽门盲囊>肝脏>肠>胃;肠的总蛋白酶活、总淀粉酶活、总脂肪酶活在各消化器官中都最高.肝脏、胃、肠、幽门盲囊的A/P值分别为2.15、22.65、3.81,1.96.研究表明,肠道是长鳍篮子鱼消化食物的最重要的消化器官;根据消化道指数、消化酶分布、A/P值,表明长鳍篮子鱼是偏植食性为主的杂食性鱼类.