Technological Science Center of Earth and Sea, UNIVALI, R Uruguai 458, Itajai, SC, 88302-202, Brazil.
Department of Marine Biodiversity Research, JAMSTEC, Natsushima-cho 2-15, Yokosuka, Kanagawa, 237-0061, Japan.
Mar Biotechnol (NY). 2017 Oct;19(5):480-487. doi: 10.1007/s10126-017-9767-3. Epub 2017 Jul 22.
Deep-sea bacteria can produce various biotechnologically relevant enzymes due to their adaptations to high pressures and low temperatures. To identify such enzymes, we have sequenced the genome of the polycaprolactone-degrading bacterium Moritella sp. JT01, isolated from sediment samples from Japan Trench (6957 m depth), using a Illumina HiSeq2000 sequencer (12.1 million paired-end reads) and CLC Genomics Workbench (version 6.5.1) for the assembly, resulting in a 4.83-Mb genome (42 scaffolds). The genome was annotated using Rapid Annotation using Subsystem Technology (RAST), Protein Homology/analogY Recognition Engine V 2.0 (PHYRE2), and BLAST2Go, revealing 4439 protein coding sequences and 101 RNAs. Gene products with industrial relevance, such as lipases (three) and esterases (four), were identified and are related to bacterium's ability to degrade polycaprolactone. The annotation revealed proteins related to deep-sea survival, such as cold-shock proteins (six) and desaturases (three). The presence of secondary metabolite biosynthetic gene clusters suggests that this bacterium could produce nonribosomal peptides, polyunsaturated fatty acids, and bacteriocins. To demonstrate the potential of this genome, a lipase was cloned an introduced into Escherichia coli. The lipase was purified and characterized, showing activity over a wide temperature range (over 50% at 20-60 °C) and pH range (over 80% at pH 6.3 to 9). This enzyme has tolerance to the surfactant action of sodium dodecyl sulfate and shows 30% increased activity when subjected to a working pressure of 200 MPa. The genomic characterization of Moritella sp. JT01 reveals traits associated with survival in the deep-sea and their potential uses in biotechnology, as exemplified by the characterized lipase.
深海细菌由于适应高压和低温,能够产生各种具有生物技术相关性的酶。为了鉴定这些酶,我们使用 Illumina HiSeq2000 测序仪(1210 万对端读长)对从日本海沟(水深 6957 米)沉积物样本中分离出的聚己内酯降解菌 Moritella sp. JT01 的基因组进行了测序,并使用 CLC Genomics Workbench(版本 6.5.1)进行组装,得到一个 4.83-Mb 的基因组(42 个支架)。使用 Rapid Annotation using Subsystem Technology (RAST)、Protein Homology/analogY Recognition Engine V 2.0 (PHYRE2) 和 BLAST2Go 对基因组进行注释,共预测到 4439 个蛋白编码序列和 101 个 RNA。鉴定出具有工业相关性的基因产物,如脂肪酶(三个)和酯酶(四个),这些基因产物与细菌降解聚己内酯的能力有关。注释结果还揭示了与深海生存相关的蛋白质,如冷休克蛋白(六个)和去饱和酶(三个)。次生代谢产物生物合成基因簇的存在表明,这种细菌可能产生非核糖体肽、多不饱和脂肪酸和细菌素。为了展示该基因组的潜力,我们克隆并引入了一个脂肪酶到大肠杆菌中。该脂肪酶经纯化和表征,在较宽的温度范围(20-60°C 之间超过 50%)和 pH 范围(pH 6.3 到 9 之间超过 80%)内具有活性。该酶对十二烷基硫酸钠的表面活性剂作用具有耐受性,在 200 MPa 的工作压力下,其活性增加了 30%。Moritella sp. JT01 的基因组特征揭示了与深海生存相关的特征及其在生物技术中的潜在用途,所鉴定的脂肪酶就是一个例证。