Distribution of Heterotrophic Bacteria in Lake Shira

A study of the horizontal and vertical distribution of heterotrophic bacteria in brackish Lake Shira in summer periods showed that mesophilic bacteria dominated in all areas of the lake, whereas psychrotolerant bacteria dominated in the metalimnion and hypolimnion of its central part. Nonhalophilic bacteria were mostly mesophilic and dominated in coastal waters. Most psychrotolerant bacteria were able to grow in the presence of 5–10% NaCl. Heterotrophic bacteria isolated in different regions of the lake were identified to a generic level. The isolates were classified into autochthonous and allochthonous microorganisms on the bases of their distribution pattern in the lake water, halotolerance, and ability to grow at low temperatures.     

基因强化在难降解污染物生物处理和修复中的应用

基因强化通过强化降解基因在土著菌群中的水平迁移和传播, 促进土著降解菌群的进化, 改善基因工程菌生物强化作用的稳定性, 提高难降解污染物的生物去除效果。介绍了基因强化的原理-微生物群落内水平基因迁移, 讨论了基因载体、细胞接触条件和环境条件等影响基因强化的因素, 综述了目前基因强化在土壤生物修复和废水生物处理中的应用现状, 并提出了基因强化中存在的问题。     

Identification of a novel 4-hydroxyphenylpyruvate dioxygenase from the soil metagenome

4-Hydroxyphenylpyruvate dioxygenase (HPPD) is a Fe(II)-dependent, non-heme oxygenase that converts 4-hydroxyphenylpyruvate to homogentisate. Essential cofactors, such as plastoquinone and tocopherol, are produced by HPPD-dependent anabolic pathways in plants. To isolate a novel hppd using culture-independent method, a cosmid metagenomic library was constructed from soil in Korea. Screening of Escherichia coli metagenomic libraries led to the identification of a positive clone, YS103B, producing dark brown pigment in Luria-Bertani medium supplemented with l-tyrosine. In vitro transposon mutagenesis of YS103B showed that the 1.3 kb insert was sufficient to produce the hemolytic brown pigment. Sequence analysis of YS103B disclosed one open reading frame encoding a 41.4 kDa protein with the well-conserved prokaryotic oxygenase motif of the HPPD family of enzymes. The HPPD-specific β-triketone herbicide, sulcotrione, inhibited YS103B pigmentation. The recombinant protein expressed in E. coli generated homogentisic acid. Thus, we present the successful heterologous expression of a previously uncharacterized hppd gene from an uncultured soil bacterium.     

原核微生物的多样性

微生物是一群以分解代谢为主的重要生物类群,其生物学多样性十分丰富。但由于它们的微观性,尤其原核微生物简单的单细胞结构、以无性方式进行快速地繁殖而造成的无准确的基线难以对其进行种群数目和数量的统计,因而对微生物的多样性研究远没有宏观生物那样深入和受到重视。本文根据原核微生物的特性,从其物种、所代表的进化分支、生理代谢类群及遗传背景几个方面简述了它们的多样性及重要意义,意在引起科学界和全社会对这类生物资源的认识和保护的重视。     

Stability of Recombinant Plasmids in Transgenic Microorganisms under Different Environmental Conditions

The copy number of R plasmids weakly depends on the selective pressure of the respective antibiotic but does depend on the physiology of the host species and the type of plasmids and cloned genes, whose expression leads to a further load on the biosynthetic apparatus of cells. The last factor is critical in the maintenance of recombinant plasmids in transgenic microorganisms.     

Distribution of Microorganisms in the Al–Fe–Humus Podzols of Natural and Anthropogenically Impacted Boreal Spruce Forests

Natural and anthropogenically induced seasonal variations in the abundance and biomass of various groups of microorganisms in the Al–Fe–humus podzols of boreal spruce forests were analyzed. The fungal biomass in these soils was found to be considerably higher than the bacterial biomass. The microbial population was mainly concentrated in a thin surface layer (10–15 cm in thickness), which included the forest litter and the upper mineral root-inhabited soil horizon and greatly differed from other soil horizons in morphology and other properties. This layer was found to be optimal with respect to hydrothermal and nutritional conditions and is characterized by profound seasonal variations in the abundance and biomass of microbiota. The high acidity, typical of the Al–Fe–humus podzols, resulted from the metabolism of their microbial communities. In the polluted podzols, the population of prokaryotes increased and that of eukaryotes decreased.     

环境样品中DNA的分离纯化和文库构建

采用研磨 /冻融和SDS/蛋白酶K热处理等理化方法 ,直接从性质不同的环境样品中提取和纯化混合基因组DNA。所获得纯品DNA的产量为每克样品 2～ 1 6μg。对纯品DNA进行限制性内切酶处理后 ,构建了以pUC1 8为载体的DNA文库。建库效率为从每克环境样品获得约 1 0 3～ 1 0 4 个含 3～ 8kb外源随机插入片段的克隆。通过DNA序列测定和基因注释 ,对从文库中随机选取的克隆进行了分析 ,发现外源插入片段均含序列未见报道的新基因。本文所做的尝试对于保存、研究和开发未培养微生物基因资源具有意义     

生物合成谷胱甘肽种间耦合ATP再生系统的构建

利用重组大肠杆菌Ⅱ 1中的谷胱甘肽合成酶系和面包酵母WSH J7中的ATP生物合成酶系 ,构建了一个以葡萄糖为能源的种间耦合ATP再生系统。经过通透性处理的酵母细胞几乎不能消耗葡萄糖。在反应体系中添加 1mmol/LAMP和 0 0 5mmol/LNADH ,即可启动酵母的酵解途径。提高耦合系统中的葡萄糖浓度 ,可促进GSH的合成。当葡萄糖浓度为 40 0mmol/L时 ,系统内GSH浓度达到 1 0 4mmol/L(3 2 g/L)。Mg2 +缺乏时 ,耦合系统和外加ATP的非耦合系统均不能合成谷胱甘肽。耦合系统中Mg2 +与ATP形成螯合物 ,可能是导致耦合系统中GSH产量较低的原因。在耦合系统中补加Mg2 +,反应 6h时GSH浓度达到 1 4 3mmol/L(4 4g/L)。     

Influence of the Degree of Aeration on Halotolerance of Yeasts of the Genera Candida,Rhodotorula, and Malassezia

The biochemical mechanisms were studied that determine different reactions of yeasts of different genera to two simultaneously imposed stressors, hypoxia and osmotic shock. For Candida lipolytica, these two stressors were antagonistic, which resulted in stimulation (and not suppression) of the growth of this yeast by NaCl (in a wide range of concentrations) under microaerobic conditions. The reaction of Malasseziasp. was different: the degree of halotolerance of this microorganism was lower under microaerobic conditions. An intervening reaction pattern was characteristic of Rhodotorula aurantiaca.These differences were found to be determined, above all, by the induction of a salt-resistant respiratory system (oxidase) in C. lipolytica, which could not be induced in Malassezia sp. In addition, the synthesis of catalase was enhanced in C. lipolytica, which provided for neutralization of the active forms of oxygen accumulating as a result of inhibition of other protective enzymes by salt.