Journalartikel

Jahr der Veröffentlichung: 1996

Seiten: 294-300

Zeitschrift: European Journal of Biochemistry

Bandnummer: 236

Heftnummer: 1

ISSN: 0014-2956

Open Access Status: Bronze

DOI Link: https://doi.org/10.1111/j.1432-1033.1996.00294.x

Verlag: Wiley: No OnlineOpen

Abstract: 
The gene mch encoding N-5,N-10-methenyltetrahydromethanopterin cyclohydrolase (Mch) in Methanobacterium thermoautotrophicum (strain Marburg) was cloned and sequenced. The gene, 963 bp, was found to be located at the 3' end of a 3.5-kbp BamHI fragment. Upstream of the mch gene two open reading frames were recognized, one encoding for a 25-kDa protein with sequence similarity to deoxyuridylate hydroxymethylase and the other encoding for a 34.6-kDa protein with sequence similarity to cobalamin-independent methionine synthase (MetE). The N-terminal amino acid sequence deduced for the deoxyuridylate hydroxymethylase was identical to that previously published for thymidylate synthase (TysY) from M. thermoautotrophicum. The 3' end of the tysY gene overlapped by 8 bp with the 5' end of the mch gene. Despite this fact, the mch gene appeared to be transcribed monocistronically as evidenced by Northern blot analysis and primer-extension experiments. The mch gene was overexpressed in Escherichia coli yielding an active enzyme of 37 kDa with a specific activity of 30 U/mg cell extract protein.

Harvard-Zitierstil: Vaupel, M., Dietz, H., Linder, D. and Thauer, R. (1996) Primary structure of cyclohydrolase (Mch) from Methanobacterium thermoautotrophicum (strain Marburg) and functional expression of the mch gene in Escherichia coli, European Journal of Biochemistry, 236(1), pp. 294-300. https://doi.org/10.1111/j.1432-1033.1996.00294.x

APA-Zitierstil: Vaupel, M., Dietz, H., Linder, D., & Thauer, R. (1996). Primary structure of cyclohydrolase (Mch) from Methanobacterium thermoautotrophicum (strain Marburg) and functional expression of the mch gene in Escherichia coli. European Journal of Biochemistry. 236(1), 294-300. https://doi.org/10.1111/j.1432-1033.1996.00294.x