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.

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

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	-	-

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	-	in terms of activity, the three constructs (tagless OcDH, His5-tagged OcDH and His6-tagged OcDH) are indistinguishable	Pecten maximus

Synonyms

Synonyms	Comment	Organism
OcDH	-	Pecten maximus
Octopine dehydrogenase	-	Pecten maximus

Cofactor

Cofactor	Comment	Organism	Structure
NADH	-	Pecten maximus