南大洋深海嗜冷菌2-5-10-1及其低温脂肪酶的研究

从南大洋普里兹湾的水样中筛选到一株产低温脂肪酶的深海嗜冷菌2-5-10-1,对其生长及产酶情况和酶性质做了初步研究.该菌株的最适生长温度为5℃,此时分泌的胞外脂肪酶最多;添加Tween80,橄榄油可显著促进脂肪酶的产生.该脂肪酶的最适作用温度为35℃,在0～20℃均保持较高的酶活性,在0℃可保持37%的相对酶活性;酶的最适作用的pH值为7.5,在pH6～9的范围内均存在较高酶活性;对热较敏感,在60℃保温15min可丧失50%以上的酶活性.该脂肪酶的催化作用不需要金属离子的参与,Cu2+和Zn2+对酶活有着强烈的抑制作用.     

Transposon mutagenesis of Psychrobacter cryohalolentis PAMC 21807 by tri-parental conjugation

Random mutagenesis is commonly used to study gene function. The screening of mutants exhibiting specific pheno- types assists in the identification of phenotype-related genes. In the current study, we isolated Antarctic bacteria, and developed a transposon Tn5 mutagenesis system. A total of 26 strains were isolated from seawater and freshwater near Antarctic King Sejong Research Station, King George Island. Six Psychrobacter strains were identified as psychrophilic, with optimal growth tempera- tures of 10~C or 15~C Psychrobacter cryohalolentis PAMC 21807 with a high growth rate at 4~C was selected for transposon mutagenesis. Tri-parental conjugation with a plasmid containing Tn5 produced 13 putative recombinants containing the selectable marker. Genomic Southern hybridization confirmed Tn5 existed as episomes for seven recombinants, and for a single recombinant, Tn5 was integrated into the genome of Psychrobacter cryohalolentis PAMC 21807. The result indicates that the mutagenesis method, although successful, has a relatively low rate. The psychrophilic bacteria isolated in this study may be a useful resource for studying cold adaptation mechanisms, and the mutagenesis method can be applied to genetic analysis.     

Study on low-temperature lipase of psychrophilic bacterium 2-5-10-1 isolated from deep sea of Southern Ocean

A strain of psychrophilic bacterium, 2-5-10-1, which produces low-temperature lipase, is isolated from the deep sea of Prydz Bay in Southern Ocean. The highest lipase secretion of this strain is observed at 5 ℃ and this temperature is also for optimal growth. Tween 80 and olive oil enhance secretion of lipase. The optimal temperature and pH for lipase activity are 35 ℃ and 7.5 ℃ respectively. At 0℃, the lipase still has 37% relative enzyme activity. The lipase shows high thermolability, more than 50% activity lost after incubation at 60 ℃ for 15 min. EDTA has no effect on lipase activity, indicating the lipase activity is independent of divalent cation. In contrast, the lipase activity is inhibited drastically by Cu2 and Zn2 .     

一株深海嗜低温萘降解细菌Nah-1的分离及降解基因研究

以萘为唯一碳源和能源从南,中绳海槽深海沉积物中分离得到一株能降解萘的海洋嗜低温细菌Nah-1,测定了该茵的最适生长条件及生长曲线。16S rRNA基因（16SrDNA）序列同源性分析表明该菌属于解环菌属（Cyczocznsticus）。PCR扩增萘降解基因得到目标片段,比对结果表明,相似度最高的基因phnAl来自Cycloclasticus sp．A5,为99％,该基因编码的蛋白是萘双加氧酶大亚基。     

南极石油烃降解嗜冷菌的筛选及其降解特性的研究

从385株南极海洋细菌中筛选出2株石油烃降解菌NJ276和NJ341,并对其降解特性进行了初步研究。以柴油为唯一碳源进行降解实验的结果表明,它们在5℃时20 d内对柴油的降解率分别达到23.47%和32.15%,在15℃时20 d内降解率分别达到43.95%和62.47%,其降解能力随着培养温度的升高而显著增强;石油烃降解残油组分的GC-MS分析结果表明,柴油经过嗜冷菌NJ276降解后的残油组分中能检测到C15～C21七种烷烃,经过嗜冷菌NJ341降解后的残油组分只能检测到少量C16,C17和C18三种烷烃。对它们进行16S rDNA基因序列的同源性和系统发育分析结果表明,菌株NJ276属于假交替单胞菌属(Pseudoaltero monas),菌株NJ341属于科尔韦尔氏属(Colwellia)。     

南极产低温几丁质酶菌株的筛选及分子鉴定

从南极普里兹湾深海底泥样品中分离得到一批嗜冷细菌,对这些嗜冷细菌进行了产几丁质酶菌株的筛选,对其中一株具有较高产酶活性的菌株AC167所分泌的几丁质酶进行了性质分析,并通过16S rDNA序列分析进行了菌株的分子鉴定。该菌株最适生长温度为10℃,最高生长温度为25℃,是典型的嗜冷菌,经16S rDNA序列比较及系统发育分析鉴定为假单胞菌属(Pseudomonas)。该菌株只在低于25℃的条件下分泌几丁质酶,酶的最适反应温度为30°C,属于低温酶。