Wackett Lawrence P, Frias Janice A, Seffernick Jennifer L, Sukovich David J, Cameron Stephan M
Department of Biochemistry, Molecular Biology and Biophysics, University of Minnesota, St. Paul, MN 55108, USA.
Appl Environ Microbiol. 2007 Nov;73(22):7192-8. doi: 10.1128/AEM.01785-07. Epub 2007 Oct 5.
Vibrio furnissii M1 was recently reported to biosynthesize n-alkanes when grown on biopolymers, sugars, or organic acids (M. O. Park, J. Bacteriol. 187:1426-1429, 2005). In the present study, V. furnissii M1 was subjected to genomic analysis and studied biochemically. The sequence of the 16S rRNA gene and repetitive PCR showed that V. furnissii M1 was not identical to other V. furnissii strains tested, but the level of relatedness was consistent with its assignment as a V. furnissii strain. Pulsed-field gel electrophoresis showed chromosomal bands at approximately 3.2 and 1.8 Mb, similar to other Vibrio strains. Complete genomic DNA from V. furnissii M1 was sequenced with 21-fold coverage. Alkane biosynthetic and degradation genes could not be identified. Moreover, V. furnissii M1 did not produce demonstrable levels of n-alkanes in vivo or in vitro. In vivo experiments were conducted by growing V. furnissii M1 under different conditions, extracting with solvent, and analyzing extracts by gas chromatography-mass spectrometry. A highly sensitive assay was used for in vitro experiments with cell extracts and [(14)C]hexadecanol. The data are consistent with the present strain being a V. furnissii with properties similar to those previously described but lacking the alkane-producing phenotype. V. furnissii ATCC 35016, also reported to biosynthesize alkanes, was found in the present study not to produce alkanes.
最近有报道称,弗氏弧菌M1在以生物聚合物、糖类或有机酸为碳源生长时能够生物合成正构烷烃(M. O. Park,《细菌学杂志》187:1426 - 1429,2005年)。在本研究中,对弗氏弧菌M1进行了基因组分析并开展了生化研究。16S rRNA基因序列和重复PCR结果表明,弗氏弧菌M1与所测试的其他弗氏弧菌菌株并不相同,但其亲缘关系水平与其作为弗氏弧菌菌株的分类一致。脉冲场凝胶电泳显示其染色体条带大小约为3.2和1.8 Mb,与其他弧菌菌株相似。对弗氏弧菌M1的完整基因组DNA进行了测序,测序深度为21倍覆盖度。未鉴定到烷烃生物合成和降解基因。此外,弗氏弧菌M1在体内和体外均未产生可检测水平的正构烷烃。体内实验是通过在不同条件下培养弗氏弧菌M1,用溶剂提取,然后通过气相色谱 - 质谱联用仪分析提取物来进行的。体外实验使用细胞提取物和[(14)C]十六醇进行了高灵敏度检测。这些数据表明,本菌株是一株弗氏弧菌,其特性与先前描述的相似,但缺乏产烷烃表型。本研究还发现,同样报道过能生物合成烷烃的弗氏弧菌ATCC 35016也不产烷烃。