Whole-genome sequencing of a psychrophilic bacterium, analysis of genes involved in cold adaptation, and exploitation of cold-active enzymes

1. 我们对希瓦氏菌进行了全基因组分析。 Ac10，一种从南极海洋分离出来的嗜冷菌。基因组大小为4.5 Mbp，ORF数约为4, 800.2。希瓦氏菌属Ac10在4℃下生长，产生大量与大肠杆菌冷休克蛋白CspA同源的蛋白。3．希瓦氏菌 (Shewanella sp.) 的胶原酶。 Ac1O的过量产生和表征。4．我们建立了希瓦氏菌的转化方法。 Ac1O，分离启动子，构建表达载体。 5．一种耐冷细菌，不动杆菌属。菌株号6、从西伯利亚苔原土壤中分离出来。该细菌产生细胞外脂肪分解酶，并在低温下降解培养基中的各种甘油三酯。该菌有望用于低温废水处理。 6．从不动杆菌属中克隆了编码具有脂肪酸乙烯基酯底物偏好的冷活性脂肪分解酶的基因。菌株号6. 该酶被证明可用于使用乙烯基酯作为酰基供体通过酯交换反应合成酯。7．我们从不动杆菌属中克隆了编码冷活性酯酶的基因。菌株号6属于酯酶/脂肪酶家族的EST组。8．我们构建了一个载体，将外源基因引入不动杆菌属。菌株号6、分离出适合外源基因过表达的强大启动子。

1. We carried out whole-genome analysis of Shewanella sp. Ac10, a psychrophile isolated from the Antarctic Ocean. The genome size was 4.5 Mbp, and the number of ORFs was about 4, 800.2. Shewanella sp. Ac10 grown at 4℃ produced a large quantity of a protein homologous to a cold-shock protein, CspA, from Escherichia coli.3. A collagenase of Shewanella sp. Ac1O was overproduced and characterized.4. We established a transformation method for Shewanella sp. Ac1O, isolated a promoter, and constructed an expression vector.5. A psychrotrophic bacterium, Acinetobacter sp. strain no. 6, was isolated from Siberian tundra soil. The bacterium produced extracellular lipolytic enzymes, and degraded various triglycerides in the medium at low temperatures. The bacterium is expected to be useful for the waste water treatment at low temperatures.6. The gene coding for a cold-active lipolytic enzyme with a substrate preference for fatty acid vinyl esters was cloned from Acinetobacter sp. strain no. 6. The enzyme was shown to be useful for the synthesis of esters by transesterification using vinyl esters as an acyl donor.7. We cloned the gene coding for a cold-active esterase from Acinetobacter sp. strain no. 6 that belongs to the EST group of the esterase/lipase family.8. We constructed a vector to introduce exogenous genes into Acinetobacter sp. strain no. 6 and isolated powerful promoters suitable for overexpression of exogenous genes.