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N 5,N 10-Methenyltetrahydromethanopterin cyclohydrolase from the extremely thermophilic sulfate reducing Archaeoglobus fulgidus: comparison of its properties with those of the cyclohydrolase from the extremely thermophilic Methanopyrus kandleri

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Abstract

Archaeoglobus fulgidus and Methanopyrus kandleri are both extremely thermophilic Archaea with a growth temperature optimum at 83°C and 98°C, respectively. Both Archaea contain an active N 5,N 10-methenyltetrahydromethanopterin cyclohydrolase. The enzyme from M. kandleri has recently been characterized. We describe here the purification and properties of the enzyme from A. fulgidus.

The cyclohydrolase from A. fulgidus was purified 180-fold to apparent homogeneity and its properties were compared with those recently published for the cyclohydrolase from M. kandleri. The two cytoplasmic enzymes were found to have very similar molecular and catalytic properties. They differed, however, significantly with respect of the effect of K2HPO4 and of other salts on the activity and the stability. The cyclohydrolase from A. fulgidus required relatively high concentrations of K2HPO4 (1 M) for optimal thermostability at 90°C but did not require salts for activity. Vice versa, the enzyme from M. kandleri was dependent on high K2HPO4 concentrations (1.5 M) for optimal activity but not for thermostability. Thus the activity and structural stability of the two thermophilic enzymes depend in a completely different way on the concentration of inorganic salts. The molecular basis for these differences are discussed.

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Abbreviations

H4MPT:

tetrahydromethanopterin

MFR:

methanofuran

CH3−H4MPT:

N 5-methyl-H4MPT

CH2=H4MPT:

N 5,N 10-methylene-H4MPT

CH2≡H4MPT:

N 5,N 10-methenyl-H4MPT

CHO−H4MPT:

N 5 formyl-H4MPT

CHO-MFR:

formyl-MFR

cyclohydrolase:

N 5,N 10-methenyltetrahydromethanopterin cyclohydrolase

MOPS:

3-(N-morpholino) propane sulfonic acid

TRICINE:

N-tris (hydroxymethyl) methyl glycine

1 U=1:

μmol/min

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Authors and Affiliations

  1. Laboratorium für Mikrobiologie des Fachbereichs Biologie der Philipps-Universität, W-3550, Marburg/Lahn, Germany

    A. R. Klein, J. Breitung & R. K. Thauer

  2. Max-Planck-Institut für Terrestrische Mikrobiologie, W-3550, Marburg/Lahn, Germany

    A. R. Klein, J. Breitung & R. K. Thauer

  3. Fachbereich Humanmedizin, Biochemisches Institut, Justus-Liebig-Universität Giessen, W-6300, Giessen, Germany

    D. Linder

  4. Lehrstuhl für Mikrobiologie, Universität Regensburg, Universitätsstrasse 31, W-8400, Regensburg, Germany

    K. O. Stetter

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  1. A. R. Klein
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  2. J. Breitung
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  4. K. O. Stetter
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Klein, A.R., Breitung, J., Linder, D. et al. N 5,N 10-Methenyltetrahydromethanopterin cyclohydrolase from the extremely thermophilic sulfate reducing Archaeoglobus fulgidus: comparison of its properties with those of the cyclohydrolase from the extremely thermophilic Methanopyrus kandleri . Arch. Microbiol. 159, 213–219 (1993). https://doi.org/10.1007/BF00248474

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