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地中海东部深层缺氧高盐湖微生物培养物中的芳烃降解模式及儿茶酚2,3-双加氧酶基因

Aromatic hydrocarbon degradation patterns and catechol 2,3-dioxygenase genes in microbial cultures from deep anoxic hypersaline lakes in the eastern Mediterranean sea.

作者信息

Brusa T, Borin S, Ferrari F, Sorlini C, Corselli C, Daffonchio D

机构信息

Dipartimento di Scienze e Tecnologie Alimentari e Microbiologiche, Università degli Studi di Milano, Italy.

出版信息

Microbiol Res. 2001;156(1):49-58. doi: 10.1078/0944-5013-00075.

Abstract

Several mixed cultures able to grow on different aromatic hydrocarbons were obtained from different depths (between 3500 and 3660 m under the sea surface) of water/brine interfaces (1 to 5 m over the estimated brine surface) of three deep hypersaline anoxic basins (Urania, Discovery and Atalante) in the eastern Mediterranean sea. Eight strains which completely removed toluene from the medium in six to 10 days were isolated from one of the mixed cultures obtained from the Urania basin. The strains grew on toluene and yeast extract in the presence of NaCl concentrations of up to 50 and 100 g l(-1), respectively, indicating that they are halotolerant rather than halophilic. Even though DNA fingerprinting methods showed that the strains were strictly related, two groups could be found on the basis of the plasmid profile. Metabolic profiling and partial sequencing (350 bp) of the 16S rDNA showed that the strains were related to Pseudomonas mendocina. A 320 bp fragment of the catechol 2,3-dioxygenase gene from all the strains was aimplified by PCR. The sequence of the fragment showed 100% identity with xylE from pWW53 of Pseudomonas putida MT53 isolated from soil. Southern hybridisation experiments showed that catechol 2,3-dioxygenase is plasmid encoded.

摘要

从地中海东部三个深度超咸缺氧盆地(乌拉尼亚、发现和阿塔兰特)不同深度(海面以下3500至3660米)的水/盐水界面(估计盐水表面上方1至5米)获得了几种能够在不同芳烃上生长的混合培养物。从乌拉尼亚盆地获得的一种混合培养物中分离出八株在6至10天内将甲苯从培养基中完全去除的菌株。这些菌株分别在NaCl浓度高达50和100 g l(-1)的情况下,以甲苯和酵母提取物为培养基生长,这表明它们是耐盐的而非嗜盐的。尽管DNA指纹图谱方法显示这些菌株密切相关,但根据质粒图谱可分为两组。16S rDNA的代谢谱分析和部分测序(350 bp)表明这些菌株与门多萨假单胞菌有关。通过PCR扩增了所有菌株儿茶酚2,3-双加氧酶基因的320 bp片段。该片段的序列与从土壤中分离的恶臭假单胞菌MT53的pWW53中的xylE显示出100%的同一性。Southern杂交实验表明儿茶酚2,3-双加氧酶是由质粒编码的。

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