Literature summary for 4.2.3.104 extracted from

Alemdar, S.; Hartwig, S.; Frister, T.; Koenig, J.C.; Scheper, T.; Beutel, S.
Heterologous expression, purification, and biochemical characterization of alpha-humulene synthase from Zingiber zerumbet Smith (2016), Appl. Biochem. Biotechnol., 178, 474-489 .

Search for more literature for 4.2.3.104

Cloned(Commentary)

Cloned (Comment)	Organism
gene ZSS1 or hum, recombinant expression of His-tagged enzyme in Escherichia coli strain BL21(DE3) under control of the lac promoter	Zingiber zerumbet

Protein Variants

Protein Variants	Comment	Organism
additional information	alpha-humulene production using the engineered Escherichia coli BL21(DE3) strain in a 2-l bioreactor cultivation, method optimization, overview. Coordinating Cu2+ ions to the NTA ligands in affinity chromatography results in a high non-specific binding of endogenous host cell proteins due to the high affinity of Cu2+ to histidine groups. Zn2+-IMAC shows much less impurities in eluted protein fractions which is consistent with the fact that Zn2+ ions are less affine to histidine groups. The recovery rate of 80 % of the protein of interest using Zn2+ ions is the lowest compared to all other metal ions tested. Ni2+- and Co2+-IMAC seem to be similar in product purity (60% and 63%) and recovery (93% and 95%). As Co2+-IMAC shows the best purity to recovery ratio, further purification is done using Co2+-IMAC	Zingiber zerumbet

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
additional information	-	additional information	Michaelis-Menten kinetics, overview	Zingiber zerumbet
0.033	-	(2E,6E)-farnesyl diphosphate	pH 7.0, 38°C, recombinant enzyme	Zingiber zerumbet

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
(2E,6E)-farnesyl diphosphate	Zingiber zerumbet	-	alpha-humulene + diphosphate	-	?

Organism

Organism	UniProt	Comment	Textmining
Zingiber zerumbet	B1B1U3	-	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3)by cobalt affinity chromatography, dialysis, and ultrafiltration, to homogeneity	Zingiber zerumbet

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
(2E,6E)-farnesyl diphosphate	-	Zingiber zerumbet	alpha-humulene + diphosphate	-	?
additional information	the recombinant alpha-humulene synthase produces 93.45% alpha-humulene and 5.5% beta-caryophyllene. Product determination by gas chromatography-flame ionization detection (GC-FID)	Zingiber zerumbet	?	-	?

Subunits

Subunits	Comment	Organism
?	x * 64500, about, sequence calculation	Zingiber zerumbet

Synonyms

Synonyms	Comment	Organism
hum	-	Zingiber zerumbet
ZSS1	-	Zingiber zerumbet

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
38	-	recombinant His-tagged enzyme	Zingiber zerumbet

Temperature Range [°C]

Temperature Minimum [°C]	Temperature Maximum [°C]	Comment	Organism
32	44	activity range, profile overview, recombinant His-tagged enzyme	Zingiber zerumbet

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism	Structure
3.8	-	(2E,6E)-farnesyl diphosphate	pH 7.0, 38°C, recombinant enzyme	Zingiber zerumbet

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
7.5	-	recombinant His-tagged enzyme	Zingiber zerumbet

pH Range

pH Minimum	pH Maximum	Comment	Organism
6	9	activity range, profile overview, recombinant His-tagged enzyme	Zingiber zerumbet

pI Value

Organism	Comment	pI Value Maximum	pI Value
Zingiber zerumbet	recombinant His-tagged enzyme, isoelectric focusing	-	5.61
Zingiber zerumbet	sequence calculation	-	5.65

General Information

General Information	Comment	Organism
metabolism	the enzyme is involved in the zerumbone biosynthesis in Zingiber zerumbet, proposed pathway and mechanism, overview	Zingiber zerumbet
physiological function	alpha-humulene is a common sesquiterpene widely distributed in plants, the synthase from Zingiber zerumbet is the only known sesquiterpene synthase to date catalyzing the formation of alpha-humulene as major product	Zingiber zerumbet

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Release 2023.1 (January 2023)