The first archaeal ATP-dependent glucokinase, from the hyperthermophilic crenarchaeon Aeropyrum pernix, represents a monomeric, extremely thermophilic ROK glucokinase with broad hexose specificity

DC FieldValueLanguage
dc.contributor.authorHansen, T
dc.contributor.authorReichstein, B
dc.contributor.authorSchmid, R
dc.contributor.authorSchonheit, P
dc.date.accessioned2021-12-23T16:06:51Z-
dc.date.available2021-12-23T16:06:51Z-
dc.date.issued2002
dc.identifier.issn00219193
dc.identifier.urihttps://osnascholar.ub.uni-osnabrueck.de/handle/unios/7580-
dc.description.abstractAn ATP-dependent glucokinase of the hyperthermophillic aerobic crenarchaeon Aeropyrum pernix was purified 230-fold to homogeneity. The enzyme is a monomeric protein with an apparent molecular mass of about 36 kDa. The apparent K-m values for ATP and glucose (at 90degreesC and pH 6.2) were 0.42 and 0.044 mM, respectively; the apparent V-max was about 35 U/mg. The enzyme was specific for ATP as a phosphoryl donor, but showed a broad spectrum for phosphoryl acceptors: in addition to glucose, which showed the highest catalytic efficiency (k(cat)/K-m), the enzyme also phosphorylates glucosamin, fructose, mannose, and 2-deoxyglucose. Divalent cations were required for maximal activity: Mg2+, which was most effective, could partially be replaced with Co2+, Mn2+, and Ni2+. The enzyme had a temperature optimum of at least 100degreesC and showed significant thermostabillity up to 100degreesC. The coding function of open reading frame (ORF) APE2091 (Y. Kawarabayasi, Y. Hino, H. Horikawa, S. Yamazaki, Y. Haikawa, K. Jin-no, M. Takahashi, M. Sekine, S. Baba, A. Ankai, H. Kosugi, A. Hosoyama, S. Fukui, Y. Nagai, K. Nishijima, H. Nakazawa, M. Takamiya, S. Masuda, T. Funahashi, T. Tanaka, Y. Kudoh, J. Yamazaki, N. Kushida, A. Oguchi, and H. Kikuchi, DNA Res. 6:83-101, 145-152, 1999), previously annotated as gene glk, coding for ATP-glucokinase of A. pernix, was proved by functional expression in Escherichia coli. The purified recombinant ATP-dependent glucokinase showed a 5-kDa higher molecular mass on sodium dodecyl sulfate-polyacrylamide gel electrophoresis, but almost identical kinetic and thermostability properties in comparison to the native enzyme purified from A. pernix. N-terminal amino acid sequence of the native enzyme revealed that the translation start codon is a GTG 171 bp downstream of the annotated start codon of ORF APE2091. The amino acid sequence deduced from the truncated ORF APE2091 revealed sequence similarity to members of the ROK family, which comprise bacterial sugar kinases' and transcriptional repressors. This is the first report of the characterization of an ATP-dependent glucokinase from the domain of Archaea, which differs from its bacterial counterparts by its monomeric structure and its broad specificity for hexoses.
dc.language.isoen
dc.publisherAMER SOC MICROBIOLOGY
dc.relation.ispartofJOURNAL OF BACTERIOLOGY
dc.subjectADP
dc.subjectBIOCHEMICAL-CHARACTERIZATION
dc.subjectEMBDEN-MEYERHOF
dc.subjectFAMILIES
dc.subjectGENOME SEQUENCE
dc.subjectGLUCOSE KINASE
dc.subjectHEXOKINASE
dc.subjectMicrobiology
dc.subjectPURIFICATION
dc.subjectSTRUCTURAL BASIS
dc.subjectSUGAR UTILIZATION
dc.titleThe first archaeal ATP-dependent glucokinase, from the hyperthermophilic crenarchaeon Aeropyrum pernix, represents a monomeric, extremely thermophilic ROK glucokinase with broad hexose specificity
dc.typejournal article
dc.identifier.doi10.1128/JB.184.21.5955-5965.2002
dc.identifier.isiISI:000178586800017
dc.description.volume184
dc.description.issue21
dc.description.startpage5955
dc.description.endpage5965
dc.identifier.eissn10985530
dc.publisher.place1752 N ST NW, WASHINGTON, DC 20036-2904 USA
dcterms.isPartOf.abbreviationJ. Bacteriol.
dcterms.oaStatusBronze, Green Published
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