摘要
南极地区在研究全球变化中具有突出的地位，南极地区的各种环境因子对全球气候变化有敏感的反应，而且它是地球上唯一未被人类活动大量影响的地区，所以，南极地区的生态学研究有特殊的重要性。南极企鹅在某些地区是生态系统的主要影响因子，企鹅粪便沉积物记录了上千年企鹅数量和分布的变化规律，从而可以反映气候、环境变化和环境污染、人类活动等因子对生态系统的影响。对追溯南极大陆生态、气候与环境演变、探讨南极环境演变与全球变化的关系都具有重要的科学意义。
本文作者于2001年11月至2002年4月参加了中国第18次南极考察队，合作采集了南极乔治王岛上阿德雷岛的企鹅粪便沉积物样品，分析了沉积物中可培养微生物，企鹅粪便沉积物中微生物数量比南极其他地区多，最多达到1×10~7个／g土样，并发现第7层次的可培养微生物数量突然降低；建立了提取南极沉积物的总DNA的方法，利用PCR-RFLP方法研究了沉积物中细菌的16S rDNA多样性，分析了沉积物中各类群细菌的数量和分布，发现沉积物中的细菌多样性较高，以CFB(Cytophaga-Flexibacter-Bacterioides)类群和β-、γ-Proteobacteria亚群的细菌为主，并且在第7层次处发现细菌的16S rDNA RFLP谱型数，基因型数和CFB类群细菌数量的异常，而β-Proteobacteria亚群和CFB类群细菌的数量呈反比关系；在沉积物中发现了大量的与降解有机污染物相关的细菌以及一株与重金属污染有关的细菌，在微生物生态方面找到了南极遭受有机物污染和重金属污染的证据；研究了沉积物各层次中各种氨基酸的种类与含量，发现沉积物中氨基酸种类和含量高于一般腐殖土，其中组氨酸、色氨酸、酪氨酸、甲硫氨酸和半胱氨酸在第2cm和第7cm处含量出现异常增多，其他氨基酸则在第2cm和第7cm处含量减少。基于以上的试验，我们分析了企鹅粪便沉积物中微生物生态与环境变化和环境污染的关系并提出了沉积物中出现的异常在环境和气候方面可能的解释，初步建立了以分析企鹅粪便沉积物中微生物多样性为手段的新的考察南极气候变迁和环境污染的思路。
深海蕴藏着丰富的资源，其中深海生物资源是国内外研究的热点。深海生物经过长期的进化、选择，在各种极端的环境条件下形成了独特的组织结构、酶系统及代谢机制以进行生存和繁衍。对这些独特的生理结构及机能进行研究，探讨深海生物适应特殊的生存环境而发生的基因变异无论在科学上还是实际开发应用上都具有重大的意义。蛋白酶是一个主要酶制剂品种，其产量仅次于淀粉酶。蛋白酶在工业、农业、医药卫生、洗涤、生物制剂、食品等方面都有极其广泛的应用。
我们从深海的样品中筛选到一株可分泌低温蛋白酶的菌株DY-A。此菌株为嗜冷嗜碱细菌，经16SrRNA鉴定，属假单胞菌属。对菌株DY-A进行了初步的细菌学和生理生化方面的研究后，我们研究了其产蛋白酶的最佳条件并利用Sephadex G-100葡聚
摘要
糖分子筛和CL一6B DEAE纤维素离子交换柱纯化了酶蛋白，考察了蛋白酶的性质，发
现此酶具有低温活性，在高碱性环境中有活力，属于丝氨酸蛋白酶;提取了菌株DY一A
的总DNA并构建了它的基因组质粒文库，进行了极端微生物蛋白酶基因克隆的尝试。
通过以上的研究，该菌株有望成为进一步研究低温碱性蛋白酶的出发菌株，在基础研
究和工业生产上有较大的应用潜力。
The Antarctica plays an important role in the study of global changes and it is the only land without large-scale human activities influence which makes it is sensitive to the global climate changes. Penguin is the most impartment factor in the Antarctic environment and its dropping sediment records the history of the population and the distribution change on the Ardley Island. It is possible for us to detect the relation among ecology, environment changes, climate variance and human activities by studying the penguin dropping sediment.
The sediment core was collected from the shore on the Ardley Peninsula during the eighteenth China Antarctic Research Expedition using a 12cm-diameter PVC pipe. The number of cultural bacteria was higher than other areas of the Antarctic and there was an obvious decrease at 7th layer. The total DNA of every layer was extracted and the 16S rDNA was amplified by PCR. The diversity of 16S rDNA was studied by RFLP (Restriction Fragment Length Polymorphism) and phylogenetic analysis. In the sediment, bacteria belonging to the CFB (Cytophaga-Flexibacter-Bacterioides) group and the β-Proteobacteria, γ-Proteobacteria were the main groups. The 16S rDNA diversity of 7th layer was different from that of other layers, including the analysis of RFLP, gene type and the population of CFB group. And the population of β-Proteobacteria showed inverse ratio with that of CFB group. Fifteen percent of the total strains were related with degradation of hydrocarbon and one strain with heavy metal contamination. The contents of ami
no acid were analyzed and the amount of Cys, Met, Tyr, His and Trp increased at 2nd and 7th layers while other kinds of amino acid decreased. The relationship between microbiology and environment changes and human activities influence was assumed based on these results.
More than 50% of the earth's surface is covered by deep-sea area. Deep-sea microorganism has especial histiocyte system, metabolism and enzyme system in long-term evolution in deep-sea environment. And the microbial proteases are among the most important hydrolytic enzymes and have been studied extensively since the advent of enzymology. The bulk uses of alkaline proteases in industrial sectors are including food and feed industry, peptide synthesis, leather industry, management of industrial and household waste, photographic industry.
A strain named DY-A which produces protease was isolated from deep-sea sediment. It was identified as Pseudomonas spp. with 16S rDNA sequence and phylogenetic analysis. It can grow at a wide range of pH and salt concentration and best at 10.C. The strain produced maximum protease activity after growth at 10.C, initial pH 10.0 and inoculation volume 0.5% for 48~72h on a shaker by speed of 200r/min. Protease was purified by Sephadex G-100 and CL-6B DEAE Sepharose. The optimum pH and temperature for the protease activity are pH 10.0 and 40.C. The enzyme is cold-active alkaline serine protease. Some other properties of
the strain and the protease were discussed in detail. Total DNA of strain DY-A was extracted and clone of the protease gene was in progress
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