Literature summary for 1.5.1.11 extracted from

  • Smits, S.H.; Mueller, A.; Grieshaber, M.K.; Schmitt, L.
    Coenzyme- and His-tag-induced crystallization of octopine dehydrogenase (2008), Acta Crystallogr. Sect. F, 64, 836-839.
    View publication on PubMedView publication on EuropePMC

Application

Application Comment Organism
molecular biology His-tag-induced crystallization of OcDH is found to be dependent on the length of the His tag Pecten maximus

Cloned(Commentary)

Cloned (Comment) Organism
OcDH is cloned and expressed tagless (OcDH-tagless), as a His5-tagged OcDH (OcDH-His5) and as a His6-tagged OcDH variant (OcDHLEHis6) in Escherichia coli. The His tag is always placed at the C-terminus Pecten maximus

Crystallization (Commentary)

Crystallization (Comment) Organism
His-tag-induced crystallization of OcDH is found to be dependent on the length of the His tag. OcDH-tagless and His6-tagged OcDH do not yield crystals. The purified His5-tagged OcDH yields small crystals. Together with the coenzyme NADH the crystals obtained are single and diffract to 2.1 A resolution. Two distinct domains can be found in OcDH: an NADH-dependent glycerol-3-phosphate dehydrogenase-like domain (domain I) and an octopine dehydrogenase specific domain (domain II; Interpro database). Each domain comprises approximately half of the protein, with domain I containing the classic Rossman fold of dinucleotide-binding proteins Pecten maximus

Organism

Organism UniProt Comment Textmining
Pecten maximus Q9BHM6
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Purification (Commentary)

Purification (Comment) Organism
His5-tagged OcDH and His6-tagged OcDH are purified by Ni-NTA chromatography, yielding almost 20 mg homogenous enzyme per litre of cell culture (purity greater than 98%). The purification of OcDH-tagless requires several chromatographic steps and yields 3-5 mg per litre of cell culture, with a purity of more than 98% Pecten maximus

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
additional information
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in terms of activity, the three constructs (tagless OcDH, His5-tagged OcDH and His6-tagged OcDH) are indistinguishable Pecten maximus

Synonyms

Synonyms Comment Organism
Octopine dehydrogenase
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Pecten maximus
OcDH
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Pecten maximus

Cofactor

Cofactor Comment Organism Structure
NADH
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Pecten maximus