Screening and molecular classification of low-temperature protease from Antarctic microorganism and its characterization

107 strains producing protease were screened from 260 strains of Antarctic psychrophilic bacteria, among which proteolytic activity of five strains was more than 45 U ml^-1. The 16S rRNA gcne sequences homology and phylogcnetic analysis of five Antarctic psychrophillc bacteria showed that NJ276, NJS-9, NJ16-70,NJ345 belonged tO the described genus Pseudoalteromonas and NJ341 belonged to the genus Colwellia. The growth and the protease characteristic of four Antarctic psychrophilic bacteria had been studied, and the result showed that the 6ptimal temperature for growth and protease-produeing of four strains was about 10℃. Their growth and protease-produeing were still high during incubatlng 2-5 days. The maximum proteolytic activity occurred at pH 9 for four Antarctic psychrophilic bacteria. The optimal temperature of protease action of both strains NJ276 and NJ5-9 was about 50℃, however, the optimal temperature of protease aetlon of both strains NJ341 and NJ345 was about 40 ℃, and their proteolytic activity under 0℃ exhibited nearly 30% of the maximum activity, but their thermal stabilities were weaker. These results indicated that proteases from NJ341 and NJ345 were low-temperature proteases.     

东海大陆架异养细菌的生态分布

从海洋物质循环的观点来看,异养细菌在分解有机物质和无机化过程中起着极重要的作用。从海洋食物链的角度来看,分解各种类型有机物而得以增殖的异养细菌自身也是海洋原生动物、浮游动物及底栖动物的营养源。异养细菌在海洋生态系中占有极重要的位置。对海洋中某些细菌的代谢活动与其他海洋生物之间的相互关系的研究日益受到普遍的重视。 海洋异养细菌的生态分布资料已有不少报道。但至今尚未有人发表过东海大陆架海域海洋微生物生态调查的报告。东海大陆架海域江河交汇人海,大陆对海洋的影响极为突出,水文和底质情况复杂,水产资源丰富,因而特别富有多样化的有机物来源。调查和了解这一海域中异养细薗生态分布规律,并研究其参与物质转化过程的特异性,不仅有助于深入研究海洋生态学问题,而且对于阐述东海大陆架的特点也极有价值。本文着重讨论异养细菌在东海大陆架的生态分布特征及菌群组成的特点。     

凡纳滨对虾生态养殖新方法的研究

对凡纳滨对虾的养殖采用新的投料和管理方法,不施肥,从放苗后2h开始投料,并由2餐／d改为5餐／d;管理上采用不施抗生素药物的生态养殖模式。定期抽样测量对虾的体长。结果表明,实验对虾生长速度比对照塘提高20％～30％,成活率高达86．33％,比对照塘高约20％。实验证明该方法值得推广。     

海带育苗水系微生物的监测与控制

现场及实验室试验表明,近岸水中细菌丰度因时因地有较大差别,近岸水在潮周期中以高潮前后细菌量较低;海水在沉淀贮存过程中细菌丰度有明显变化,且与海水中营养物含量、贮存温度等有关;海水经沙滤后能减少细菌90％左右;回水中脱落苗进入贮水池,是贮水池中细菌数量增长10余倍的主要原因。     

Antiangiogenic activity of low-temperature lysozyme from a marine bacterium <Emphasis Type="Italic">in vivo</Emphasis> and <Emphasis Type="Italic">in vitro</Emphasis>

We extracted marine low-temperature lysozyme (MLTL), a novel lysozyme, from a marine microorganism through fermentation. Our previous study suggested that a low molecular weight (16 kDa) may exert anti-tumor activity through antiangiogenesis. In this study, we extracted a high weight (39 kDa) and investigated its antiangiogenic activity in vivo and in vitro. Using zebrafish embryos as an in vivo study model, we found that treatment with MLTL significantly inhibited the growth of subintestinal vessels (SIVs) in a dose-dependent manner and that 400 μg/ml MLTL was sufficient to block the growth of SIVs. An in vitro study conducted using human umbilical vein endothelial cells (HUVECs) revealed that MLTL suppressed the proliferation, migration and tube formation of HUVECs in a dose-dependent manner. Interestingly, assays by flow cytometry and DNA electrophoresis indicated that MLTL was able to induce apoptosis of HUVECs. Moreover, further study demonstrated that the disruption of intracellular Ca²⁺ homeostasis may play an important role in MLTL induced apoptosis of HUVECs. Taken together, the results of this study demonstrate for the first time that MLTL inhibits angiogenesis through its pleiotropic effects on vascular endothelial cells and induces apoptosis through regulation of cellular Ca²⁺ levels. The results of this study also revealed a possible mechanism underlying the antiangiogenic effect of MLTL and suggested that MLTL may be a promising new antiangiogenic agent for use in cancer therapy.     

Marine bacterium strain screening and pyrethroid insecticide-degrading efficiency analysis

A pyrethroid insecticide-degrading bacterium, strain HS-24, was isolated from an offshore seawater environment. The strain, which can degrade cypermethrin(CYP) and deltamethrin(DEL), was identified as Methylophaga sp. The optimal culture and degradation conditions for CYP and DEL by strain HS-24 is pH 7 at 28°C. Under optimum culture conditions, strain HS-24 exhibited a broad degradation concentration range of 100, 200, 400, 600, and 800 mg/L for CYP and DEL. The metabolic intermediates were analyzed by NMR, which provided strong evidence that CYP and DEL removal occurred mainly because of a biological process. The toxicity of the degradation products of strain HS-24 was studied simultaneously by measuring the light output of the luminescence bacterium. This demonstrated that the biodegradation ability of strain HS-24 significantly decreased the toxicity of CYP- and DEL-contaminated aquaculture seawater. Finally, the findings of this paper indicate that strain HS-24 is thus revealed as a biological agent for the remediation of marine aquatic environments.     

邻苯二甲酸二甲酯及其异构体的好氧微生物降解

3种苯二甲酸二甲酯异构体(邻、间和对苯二甲酸二甲酯)主要应用于化学工业,作为增塑剂和生产聚酯的原料。用邻苯二甲酸二丁酯为惟一碳源,从红树林底泥中驯化、富集、培养、分离得到的微生物对邻苯二甲酸二甲酯(Dimethylphthalate,DMP)及其异构体对苯二甲酸二甲酯(Terephthalate,DMT)和间苯二甲酸二甲酯(Isophthalate,DMI)具有较强的降解作用。此菌株16SrDNA分子生物学的鉴定为Rhodococcusruber1k。实验得出该菌能够在苯二甲酸二甲酯作为惟一碳源和能源的培养基中生长。浓度为50mg·L-1的DMP、DMI和DMT分别在6、10、11d内可以完全被降解;DMP能够在好氧条件下被该菌快速降解,生成邻苯二甲酸一甲酯(monomethylphthalate,MMP)和邻苯二甲酸(phthalicacid,PA)2种主要中间产物,最终可以完全矿化成CO2和H2O;该菌对DMI和DMT的降解速度则比DMP慢。两者的降解中间产物间苯二甲酸一甲酯(MMI)和对苯二甲酸一甲酯(MMT)却不能被Rhodococcusruber1k继续降解而在培养基中积累。结果表明苯二甲酸二甲基酯的3种异构体能够被红树林底泥中的土著微生物降解。降解速度及降解途径与底物的化学结构有密切关系。     

墨西哥湾GC238区冷泉碳酸盐岩的微结构与石化微生物特征

采于墨西哥湾GC238海底天然气渗漏区浅表层的冷泉碳酸盐岩呈结核状产出,由方解石微晶和胶结物及少量的黄铁矿构成。胶结物由直径为0.1～0.5m的方解石化的球体、卵形体、棒状体组成,充填于方解石晶体之间。冷泉碳酸盐岩结核下表面发育有由方解石化的球体、卵形体、棒状体组成的薄层,其中的一些球状集合体(约5m)断面显示发育有核和外壳的层圈结构。黄铁矿呈草莓状,也具有相似的层圈结构。这种层状结构与活体古细菌被硫酸盐还原细菌包裹的层圈结构相似。样品中所保存的球体、卵形体、棒状体及其所组成的层圈结构可能是石化的甲烷氧化古细菌和硫酸盐还原细菌。     

墨西哥湾GC238区冷泉碳酸盐岩的微结构与石化微生物特征

采于墨西哥湾GC238海底天然气渗漏区浅表层的冷泉碳酸盐岩呈结核状产出,由方解石微晶和胶结物及少量的黄铁矿构成。胶结物由直径为0.1～0.5 m的方解石化的球体、卵形体、棒状体组成,充填于方解石晶体之间。冷泉碳酸盐岩结核下表面发育有由方解石化的球体、卵形体、棒状体组成的薄层,其中的一些球状集合体（约5 m）断面显示发育有核和外壳的层圈结构。黄铁矿呈草莓状,也具有相似的层圈结构。这种层状结构与活体古细菌被硫酸盐还原细菌包裹的层圈结构相似。样品中所保存的球体、卵形体、棒状体及其所组成的层圈结构可能是石化的甲烷氧化古细菌和硫酸盐还原细菌。     

盘锦芦苇湿地土壤微生物特征分析

基于盘锦湿地生态系统野外观测站芦苇群落生长季6～9月的定位观测资料,分析了芦苇湿地土壤微生物季节动态及其与环境因子的关系。结果表明:盘锦芦苇湿地的土壤细菌、放线菌、真菌以及微生物的总数在生长季节呈现出先减少、后增加的变化趋势。湿地的土壤细菌、放线菌以及微生物的总数在8月达到最小,而真菌数量在7月达到最小。湿地土壤中,细菌数量最大,其次是放线菌,最少的是真菌。对细菌、放线菌、真菌以及微生物总数与环境因子的相关分析表明,细菌、放线菌与微生物总数主要受水分影响,而真菌则受水分与气温的协同作用影响