深海适冷菌SM9913产生的低温蛋白酶

从1855m深的深海泥样中分离纯化得到200多株分泌蛋白酶的适冷菌，其中3株产低温蛋白酶，本文对其中一株Pseudomonas sp.SM9913(P.SM9913)生长的适冷性和它产生的蛋白酶的适低温特性进行了研究。该菌株能够在0℃正常生长，其最适生长温度为15℃，最高生长温度为35℃。为一株适冷菌。该菌株所产蛋白酶的比合成速率在10℃时最高，催化酪蛋白水解的最适温度为35℃，在0℃仍具有3％的酶活力。最适pH为8．0。该蛋白酶的热稳定性很低，在40℃保温10min即丧失85％的活力，40℃时的半衰期为6min，为一典型的低温酶。抑制剂试验表明，该蛋白酶为金属蛋白酶。     

利用毛细管电泳检测适冷蛋白酶自溶的动态过程

蛋白酶由于具有催化蛋白质水解的能力,因此在蛋白酶溶液中 ,蛋白酶分子可以相互催化,造成蛋白酶降解 ,酶活降低,这种现象称为蛋白酶自溶（ａｕｔｏｌｙｓｉｓ）。许多蛋白酶都具有自溶的特点。蛋白酶的自溶对蛋白酶的研究和保存带来很多困难。因此,弄清蛋白酶自溶的特点和机制非常重要 ,这是探讨防止蛋白酶降解方法的基础。Ｋｕｍｕｒａ等［４］对荧光假单胞菌产生的蛋白酶的自溶现象进行了研究,他们发现该蛋白酶在５０℃下 ,保温２ｍｉｎ就丧失９０%的活性。利用ＨＰＬＣ分离自溶片段进行Ｎ端测序表明 ,该酶的自溶酶切位点是不确定的 ,但靠…     

基于FT-IR、GC及核磁共振的仿刺参(Apostichopus japonicus)消化道多糖结构解析研究

仿刺参(Apostichopusjaponicus)因其高营养价值并富含多种生物活性成分,具有重要的经济价值。为了充分利用仿刺参加工下脚料,提高养殖经济效益,本文采用复合蛋白酶解法提取仿刺参消化道多糖,研究了酶解温度、时间、pH及加酶量对仿刺参消化道多糖得率的影响,并通过正交实验对仿刺参消化道多糖提取工艺进行优化,利用DEAE-sepharoseFastFlow层析柱对多糖进行纯化,通过傅里叶红外光谱(FT-IR)、气相色谱(GC)及核磁共振(13C, 1H谱)对多糖的结构及组成进行分析。结果表明,复合蛋白酶在pH 7,加酶量1.2?104 U/g,温度60℃酶解6h;利用Sevage法脱蛋白提取得到的仿刺参消化道多糖纯度均一,效果最佳。通过FT-IR结果判断其为糖类化合物,含有β型葡萄毗喃糖苷键;利用气相色谱确定该多糖主要由鼠李糖、岩藻糖、阿拉伯糖、甘露糖、葡萄糖和半乳糖六种单糖组成,其摩尔比为1︰32.86︰14.56︰1.66︰0.78︰10.43,其中岩藻糖的含量最高;核磁共振综合分析结果进一步证实仿刺参消化道多糖含有β型毗喃已糖,且主要是以l, 4-糖苷键相连接。     

不同条件因子对海月水母胃丝蛋白酶活性的影响

本实验采用福林-酚法测定了海月水母(Aureliaaurita)胃丝酶粗提液中蛋白酶的活性,并系统地研究了不同温度、pH和金属离子对海月水母胃丝蛋白酶活性的影响。结果表明,海月水母胃丝蛋白酶具有很高的活性,且蛋白酶的最适温度为37℃,最适pH值为8.5。Mn~(2+)离子对海月水母蛋白酶活性具有良好的促进作用,而Cu~(2+)和Zn~(2+)对蛋白酶具有很强的抑制作用, Mg~(2+)、K~+、Ca~(2+)对蛋白酶活性没有明显影响。     

一株北极海洋细菌及其蛋白酶的生理生化特征研究

从北极海洋环境中筛选分离出一株产胞外碱性蛋白酶的革兰氏阴性杆菌 Ar/ W/ b/ 75°2 5′ N/ 1。该菌能在 7～ 30℃范围内生长 ,并且在 30℃条件下生长良好 ,但不能在 40℃条件下生长。该菌在 0 .5%～ 1 0 % Na Cl的盐浓度范围内能良好生长 ,表现出一定的嗜盐性。葡萄糖、蔗糖及可溶性淀粉等碳源物质可被该菌株利用以供生长之需 ,其中以葡萄糖最佳。氮源物质中 ,蛋白胨是最有利于菌株生长、产酶的有机氮源 ,而 NH4 NO3则是效果最好的无机氮源。对该菌蛋白酶的酶学特性研究表明 :所产胞外蛋白酶占其蛋白酶总量的 75.7%左右 ;蛋白酶反应的最适温度为40℃ ,酶活力在温度不高于 40℃条件下保持稳定 ;该菌蛋白酶具有较宽的 p H活性范围 ,在 p H7～ 1 1均表现出较高酶活力 ,其最适 p H在 8左右     

南冲绳海槽深海沉积物中度嗜盐菌系统进化及多样性分析

从南冲绳海槽MD05-2907站位的深海沉积物中分离到40株属于盐单胞菌科(Halomonadaceae)的中度嗜盐菌.耐盐度实验表明有12株最适生长盐度为30,19株为60,9株为100,多数菌在盐度高于150时生长速度迅速下降.有8株菌能产至少一种胞外蛋白酶.     

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.     

Immunotoxicity Effect of Benzo[α]Pyrene on Scallop Chlamys f arreri

The toxic effects of benzo[α]pyrene （B[α]P） at different concentrations （0.1, 0.5, 1, 2.5 and 7.5 μgL^-1） on scallop （Chlamys farreri） immune system were studied. The results showed that B[α]P had significant toxic effects on the haemocyte counts, neutral red uptake, phagocytosis, bacteriolytic and antibacterial activity （P〈0.05）, while the seawater control and acetone control had no significant differences. The haemocyte counts, neutral red uptake, phagocytosis and bacteriolytic activity in all B[α]P treatment groups as well as antibacterial activity in groups of 0.5, 1, 2.5 and 7.5 μgL^-1·B[α]P decreased significantly （P〈0.05）. Some of these indices tended to be stable on the sixth day and others on the ninth day, and the indices showed clear time- and concentration-response to B[α]E Bactenolytic activity in 0.1 μgL^-1 B[α]P treatment group and antibacterial activity in 0.1 ggLl and 0.5 μgL^-1 B[α]P treatment groups increased at the beginning of exposure and reached their peaks on day 1 and day 6, respectively. Following that, both activities decreased gradually and became stable after day 9. When all the indices reached stability, they were significantly lower than those in control group （P〈0.05）, except for antibacterial activity in 0.1 μgLl B[α]P treatment group （P〉0.05）. Thus, B[α]P has evident toxic effects on scallop immune system, which supports the view that a relationship exists between pollution and lmmunomodulation in aquatic organisms.     

南极普里兹湾沉积物中的糖类分布及意义

利用中国第18和21次南极考察获取的南极普里兹湾沉积物样品,分析了其中的有机碳和糖类物质的含量及组成,结果表明糖类和有机碳的分布受上层水体的初级生产、地形条件和水体垂直稳定度等多种因素的控制。表层沉积物中糖类物质的平均含量为3.03 mg/g,最高值为5.60mg/g,出现在湾内的毗邻陆架区。沉积物有机碳含量与表层海水叶绿素a具有良好的相关性,能够反映上层水体初级生产的变化。单糖组分的研究可以判定其生源母质,沉积物中有机质的来源主要是海洋上层生物。糖类是易被降解利用的有机质,通过糖类物质中六碳糖的比重及其垂直分布的变化可以判断出不同站位沉积速率的相对快慢。     

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.