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Titolo:
Characterization of alanine and malate dehydrogenases from a marine psychrophile strain PA-43
Autore:
Irwin, JA; Gudmundsson, HM; Marteinsson, VT; Hreggvidsson, GO; Lanzetti, AJ; Alfredsson, GA; Engel, PC;
Indirizzi:
Natl Univ Ireland Univ Coll Dublin, Dept Biochem, Dublin 4, Ireland Natl Univ Ireland Univ Coll Dublin Dublin Ireland 4 m, Dublin 4, Ireland Natl Univ Ireland Univ Coll Dublin, Conway Inst Biomol & Biomed Res, Dublin 4, Ireland Natl Univ Ireland Univ Coll Dublin Dublin Ireland 4 s, Dublin 4, Ireland Univ Iceland, Inst Biol, Microbiol Lab, Reykjavik, Iceland Univ Iceland Reykjavik Iceland Biol, Microbiol Lab, Reykjavik, Iceland Prokaria Ltd, Reykjavik, Iceland Prokaria Ltd Reykjavik IcelandProkaria Ltd, Reykjavik, Iceland Pfizer Global Res & Dev, Groton, CT USA Pfizer Global Res & Dev Groton CTUSA r Global Res & Dev, Groton, CT USA
Titolo Testata:
EXTREMOPHILES
fascicolo: 3, volume: 5, anno: 2001,
pagine: 199 - 211
SICI:
1431-0651(200106)5:3<199:COAAMD>2.0.ZU;2-3
Fonte:
ISI
Lingua:
ENG
Soggetto:
DEEP-SEA BACTERIUM; CRYSTAL-STRUCTURE; COLD ADAPTATION; GLUTAMATE-DEHYDROGENASES; ESCHERICHIA-COLI; BETA-LACTAMASE; ALPHA-AMYLASE; GENE CLONING; VIBRIO SP; PURIFICATION;
Keywords:
psychrophilic; cold-adapted; Vibrio; Shewanella; malate dehydrogenase; alanine dehydrogenase; glutamate dehydrogenase;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Agriculture,Biology & Environmental Sciences
Life Sciences
Citazioni:
40
Recensione:
Indirizzi per estratti:
Indirizzo: Engel, PC Natl Univ Ireland Univ Coll Dublin, Dept Biochem, Dublin 4, Ireland Natl Univ Ireland Univ Coll Dublin Dublin Ireland 4 4, Ireland
Citazione:
J.A. Irwin et al., "Characterization of alanine and malate dehydrogenases from a marine psychrophile strain PA-43", EXTREMOPHIL, 5(3), 2001, pp. 199-211

Abstract

Alanine dehydrogenase (AlaDH: EC 1.4.1.1), malate dehydrogenase (MDH: EC 1.1.1.37), and glutamate dehydrogenase (EC 1.4.1.2), all NAD(+) dependent, were detected in extracts from a psychrophilic bacterium, strain PA-43, isolated from a sea urchin off the Icelandic coast. Characterization tests suggested that the strain had a close relationship to Vibrio, but sequencing ofpart of the 16S rDNA gene placed the bacterium among Shewanella species ina constructed phylogenetic tree. The bacterium had an optimum growth temperature of 16.5 degreesC, and maximum dehydrogenase expression was obtained in a rich medium supplemented with NaCl. Both AlaDH and MDH were purified to homogeneity. AlaDH is a hexamer, with an approximate relative molecular mass of 260,000, whereas MDH is dimeric, with an apparent relative molecularmass of approximately 70,000, Both enzymes were thermolabile, and the optimum temperatures for activity were shifted toward lower temperatures than those found in the same enzymes from mesophiles, 37 degreesC for MDH and approximately 47 degreesC for AlaDH. The pH optima for AlaDH in the forward and reverse reactions were 10.5 and 9, respectively, whereas those for MDH were 10-10.2 and 8.8, respectively. Partial amino acid sequences, comprising approximately 30% of the total sequences from each enzyme, were determined for N-terminal, tryptic, and chymotryptic fragments of the enzymes. The AlaDH showed the highest similarity to AlaDHs from the psychrotroph ShewanellaAc10 and the mesophile Vibrio proteolyticus, whereas MDH was most similar to the MDHs from the mesophiles Escherichia coli and Haemophilus influenzae, with lower identity to the psychrophilic malate dehydrogenases from Vibrio 5710 and Photobacterium SS9.

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Documento generato il 02/04/20 alle ore 09:13:28