Cloning, expression, purification and characterization of recombinant (+)-germacrene D synthase from Zingiber officinale

Picaud, S.; Olsson, M.E.; Brodelius, M.; Brodelius, P.E.; Arch. Biochem. Biophys. 452, 17-28 (2006)

Data extracted from this reference:

Cloned(Commentary)

Commentary	Organism
expression in Escherichia coli	Zingiber officinale

Metals/Ions

Metals/Ions	Commentary	Organism	Structure
Co2+	1 mM, 33% of the activity with Mg2+	Zingiber officinale
Mg2+	maximum activity at 0.1 mM	Zingiber officinale
Mn2+	0.1 mM, 24% of the activity with Mg2+	Zingiber officinale
additional information	divalent cation absolutely required, inactive with Cu2+ or Zn2+	Zingiber officinale	-
Ni2+	0.5 mM, 6% of the activity with Mg2+	Zingiber officinale

Organism

Organism	Primary Accession No. (UniProt)	Commentary	Textmining
Zingiber officinale	Q0VHD6	-	-

Purification (Commentary)

Commentary	Organism
recombinant protein	Zingiber officinale

Substrates and Products (Substrate)

Substrates	Commentary Substrates	Literature (Substrates)	Organism	Products	Commentary (Products)	Literature (Products)	Organism (Products)	Reversibility
(2E,6E)-farnesyl diphosphate	-	677810	Zingiber officinale	(+)-germacrene D + diphosphate	products are 50.2% (+)-germacrene D, 17.1% germacrene B, plus a number of minor products	-	-	?
More	no substrate: geranyl diphosphate	677810	Zingiber officinale	?	-	-	-	-

Subunits

Subunits	Commentary	Organism
?	x * 63800, calculated, x * 60000, SDS-PAGE	Zingiber officinale

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Commentary	Organism
7.5	-	-	Zingiber officinale

pH Range

pH Minimum	pH Maximum	Commentary	Organism
6.5	9	almost completely inactive below and above	Zingiber officinale

pI Value

Organism	Commentary	pI Value Maximum	pI Value
Zingiber officinale	calculated	-	5.6

Cloned(Commentary) (protein specific)

Commentary	Organism
expression in Escherichia coli	Zingiber officinale

Metals/Ions (protein specific)

Metals/Ions	Commentary	Organism	Structure
Co2+	1 mM, 33% of the activity with Mg2+	Zingiber officinale
Mg2+	maximum activity at 0.1 mM	Zingiber officinale
Mn2+	0.1 mM, 24% of the activity with Mg2+	Zingiber officinale
additional information	divalent cation absolutely required, inactive with Cu2+ or Zn2+	Zingiber officinale	-
Ni2+	0.5 mM, 6% of the activity with Mg2+	Zingiber officinale

Purification (Commentary) (protein specific)

Commentary	Organism
recombinant protein	Zingiber officinale

Substrates and Products (Substrate) (protein specific)

Substrates	Commentary Substrates	Literature (Substrates)	Organism	Products	Commentary (Products)	Literature (Products)	Organism (Products)	Reversibility
(2E,6E)-farnesyl diphosphate	-	677810	Zingiber officinale	(+)-germacrene D + diphosphate	products are 50.2% (+)-germacrene D, 17.1% germacrene B, plus a number of minor products	-	-	?
More	no substrate: geranyl diphosphate	677810	Zingiber officinale	?	-	-	-	-

Subunits (protein specific)

Subunits	Commentary	Organism
?	x * 63800, calculated, x * 60000, SDS-PAGE	Zingiber officinale

pH Optimum (protein specific)

pH Optimum Minimum	pH Optimum Maximum	Commentary	Organism
7.5	-	-	Zingiber officinale

pH Range (protein specific)

pH Minimum	pH Maximum	Commentary	Organism
6.5	9	almost completely inactive below and above	Zingiber officinale

pI Value (protein specific)

Organism	Commentary	pI Value Maximum	pI Value
Zingiber officinale	calculated	-	5.6

See also following references to EC number 4.2.3.77 (sorted by year of publication):

No.	1st author	Pub Med	title	organims	journal	volume	pages	year	Activating Compound	Application	Cloned(Commentary)	Crystallization (Commentary)	Engineering	General Stability	Inhibitors	KM Value [mM]	Localization	Metals/Ions	Molecular Weight [Da]	Natural Substrates/ Products (Substrates)	Organic Solvent Stability	Organism	Oxidation Stability	Posttranslational Modification	Purification (Commentary)	Reaction	Renatured (Commentary)	Source Tissue	Specific Activity [micromol/min/mg]	Storage Stability	Substrates and Products (Substrate)	Subunits	Temperature Optimum [°C]	Temperature Range [°C]	Temperature Stability [°C]	Turnover Number [1/s]	pH Optimum	pH Range	pH Stability	Cofactor	Ki Value [mM]	pI Value	IC50 Value	Activating Compound (protein specific)	Application (protein specific)	Cloned(Commentary) (protein specific)	Cofactor (protein specific)	Crystallization (Commentary) (protein specific)	Engineering (protein specific)	General Stability (protein specific)	IC50 Value (protein specific)	Inhibitors (protein specific)	Ki Value [mM] (protein specific)	KM Value [mM] (protein specific)	Localization (protein specific)	Metals/Ions (protein specific)	Molecular Weight [Da] (protein specific)	Natural Substrates/ Products (Substrates) (protein specific)	Organic Solvent Stability (protein specific)	Oxidation Stability (protein specific)	Posttranslational Modification (protein specific)	Purification (Commentary) (protein specific)	Renatured (Commentary) (protein specific)	Source Tissue (protein specific)	Specific Activity [micromol/min/mg] (protein specific)	Storage Stability (protein specific)	Substrates and Products (Substrate) (protein specific)	Subunits (protein specific)	Temperature Optimum [°C] (protein specific)	Temperature Range [°C] (protein specific)	Temperature Stability [°C] (protein specific)	Turnover Number [1/s] (protein specific)	pH Optimum (protein specific)	pH Range (protein specific)	pH Stability (protein specific)	pI Value (protein specific)	Expression	General Information	General Information (protein specific)	Expression (protein specific)	KCat/KM [mM/s]	KCat/KM [mM/s] (protein specific)
699267	Nieuwenhuizen		Two terpene synthases are resp ...	Actinidia deliciosa	J. Exp. Bot.	60	3203-3219	2009	-	-	1	-	-	-	-	1	1	3	-	-	-	1	-	-	-	-	-	2	-	-	1	1	-	-	-	1	-	-	-	-	-	1	-	-	-	1	-	-	-	-	-	-	-	1	1	3	-	-	-	-	-	-	-	2	-	-	1	1	-	-	-	1	-	-	-	1	-	-	-	-	1	1
682659	Picaud		Improved conditions for produc ...	Zingiber officinale	Protein Expr. Purif.	51	71-79	2007	-	1	1	-	-	-	-	-	-	-	-	-	-	1	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	1	1	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
677810	Picaud		Cloning, expression, purificat ...	Zingiber officinale	Arch. Biochem. Biophys.	452	17-28	2006	-	-	1	-	-	-	-	-	-	5	-	-	-	4	-	-	1	-	-	-	-	-	2	1	-	-	-	-	1	1	-	-	-	1	-	-	-	1	-	-	-	-	-	-	-	-	-	5	-	-	-	-	-	1	-	-	-	-	2	1	-	-	-	-	1	1	-	1	-	-	-	-	-	-
660802	Prosser		Enantiospecific (+)- and (-)-g ...	Solidago canadensis	Arch. Biochem. Biophys.	432	136-144	2004	-	-	1	-	2	-	-	1	-	1	-	-	-	4	-	-	-	-	-	-	-	-	1	1	-	-	-	1	-	-	-	-	-	-	-	-	-	1	-	-	2	-	-	-	-	1	-	1	-	-	-	-	-	-	-	-	-	-	1	1	-	-	-	1	-	-	-	-	-	-	-	-	1	1
661493	Schmidt	-	Mechanisms of the biosynthesis ...	Solidago canadensis	Chirality	11	353-362	1999	-	-	-	-	-	-	-	-	-	-	-	-	-	1	-	-	1	1	-	-	-	-	1	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	1	-	-	-	-	1	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-