首页> 外文期刊>Phytochemistry >Isolation and characterization of a gene encoding a S-adenosyl-l-methionine-dependent halide/thiol methyltransferase (HTMT) from the marine diatom Phaeodactylum tricornutum: Biogenic mechanism of CH sub(3I emissions in oceans)
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Isolation and characterization of a gene encoding a S-adenosyl-l-methionine-dependent halide/thiol methyltransferase (HTMT) from the marine diatom Phaeodactylum tricornutum: Biogenic mechanism of CH sub(3I emissions in oceans)

机译:海洋硅藻Phaeodactylum tricornutum中编码S-腺苷-1-蛋氨酸依赖性卤化物/硫醇甲基转移酶(HTMT)的基因的分离和表征:CH sub(3I排放在海洋中)的生物机制

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摘要

Several marine algae including diatoms exhibit S-adenosyl-l-methionine (SAM) halide/thiol methyltransferase (HTMT) activity, which is involved in the emission of methyl halides. In this study, the in vivo biogenic emission of methyl iodide from the diatom Phaeodactylum tricornutum was found to be clearly correlated with iodide concentration in the incubation media. The gene encoding HTMT (Pthtmt) was isolated from P. tricornutum CCAP 1055/1, and expressed in Escherichia coli. The molecular weight of the enzyme was 29.7 kDa including a histidine tag, and the optimal pH was around pH 7.0. The kinetic properties of recombinant PtHTMT towards Cl[super]-, Br[super]-, I[super]-, [SH][super]-, [SCN][super]-, and SAM were 637.88 mM, 72.83 mM, 8.60 mM, 9.92 mM, 7.9 mM, and 0.016 mM, respectively, and were similar to those of higher-plant HTMTs, except that the activity towards thiocyanate was lower. The biogenic emission of methyl halides from the cultured cells and the enzymatic properties of HTMT suggest that the HMT/HTMT reaction is key to understanding the biogenesis of methyl halides in oceanic environments as well as terrestrial ones.
机译:包括硅藻在内的几种海洋藻类均表现出S-腺苷-1-蛋氨酸(SAM)卤化物/硫醇甲基转移酶(HTMT)活性,这与甲基卤化物的排放有关。在这项研究中,发现硅藻硅藻(Phaeodactylum tricornutum)体内碘甲烷的生物成因释放与孵化介质中碘化物的浓度明显相关。编码HTMT(Pthtmt)的基因是从玉米角腐病菌CCAP 1055/1中分离出来的,并在大肠杆菌中表达。该酶的分子量为29.7 kDa,包括一个组氨酸标签,最适pH约为7.0。重组PtHTMT对Cl ^-,Brsuper-,Isuper-,[SH] super-,[SCN] super-和SAM的动力学性质为637.88 mM,72.83 mM,分别为8.60 mM,9.92 mM,7.9 mM和0.016 mM,与高植物HTMT相似,除了对硫氰酸盐的活性较低。从培养的细胞中甲基卤的生物释放和HTMT的酶学性质表明,HMT / HTMT反应对于理解甲基溴在海洋以及陆地环境中的生物发生至关重要。

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