Department of Genetics, Faculty of Biology, University of Bucharest, Bucharest, Romania.
Department of Chemistry, Faculty of Science and Technology, UiT The Arctic University of Norway, Tromsø, Norway.
Microbiologyopen. 2021 Mar;10(2):e1182. doi: 10.1002/mbo3.1182.
Polyhydroxyalkanoates (PHAs) are biodegradable bioplastics that can be manufactured sustainably and represent a promising green alternative to petrochemical-based plastics. Here, we describe the complete genome of a new marine PHA-producing bacterium-Photobacterium ganghwense (strain C2.2), which we have isolated from the Black Sea seashore. This new isolate is psychrotolerant and accumulates PHA when glycerol is provided as the main carbon source. Transmission electron microscopy, specific staining with Nile Red visualized via epifluorescence microscopy and gas chromatography analysis confirmed the accumulation of PHA. This is the only PHA-producing Photobacterium for which we now have a complete genome sequence, allowing us to investigate the pathways for PHA production and other secondary metabolite synthesis pathways. The de novo assembly genome, obtained using open-source tools, comprises two chromosomes (3.5, 2 Mbp) and a megaplasmid (202 kbp). We identify the entire PHA synthesis gene cluster that encodes a class I PHA synthase, a phasin, a 3-ketothiolase, and an acetoacetyl-CoA reductase. No conventional PHA depolymerase was identified in strain C2.2, but a putative lipase with extracellular amorphous PHA depolymerase activity was annotated, suggesting that C2.2 is unable to degrade intracellular PHA. A complete pathway for the conversion of glycerol to acetyl-CoA was annotated, in accordance with its ability to convert glycerol to PHA. Several secondary metabolite biosynthetic gene clusters and a low number of genes involved in antibiotic resistance and virulence were also identified, indicating the strain's suitability for biotechnological applications.
聚羟基脂肪酸酯(PHAs)是可生物降解的生物塑料,可以可持续地制造,是一种有前途的绿色替代石化塑料。在这里,我们描述了一种新的海洋 PHA 生产细菌 Photobacterium ganghwense(菌株 C2.2)的完整基因组,该细菌是从黑海海滨分离出来的。这种新的分离株是耐寒的,当甘油作为主要碳源提供时,会积累 PHA。透射电子显微镜、尼罗红特异性染色通过荧光显微镜观察和气相色谱分析证实了 PHA 的积累。这是我们现在拥有完整基因组序列的唯一生产 PHA 的 Photobacterium,使我们能够研究 PHA 生产和其他次生代谢物合成途径的途径。使用开源工具获得的从头组装基因组由两个染色体(3.5、2 Mbp)和一个大型质粒(202 kbp)组成。我们确定了整个 PHA 合成基因簇,该基因簇编码了一种 I 类 PHA 合酶、一种pha 蛋白、一种 3-酮硫解酶和一种乙酰乙酰辅酶 A 还原酶。在菌株 C2.2 中未鉴定出常规的 PHA 解聚酶,但注释了一种具有细胞外无定形 PHA 解聚酶活性的假定脂肪酶,表明 C2.2 无法降解细胞内的 PHA。根据其将甘油转化为 PHA 的能力,注释了一条完整的将甘油转化为乙酰辅酶 A 的途径。还鉴定了几个次生代谢物生物合成基因簇和少数参与抗生素抗性和毒力的基因,表明该菌株适合生物技术应用。