Literature summary for 1.14.11.11 extracted from

Cao, Y.D.; He, Y.C.; Li, H.; Kai, G.Y.; Xu, J.H.; Yu, H.L.
Efficient biosynthesis of rare natural product scopolamine using E. coli cells expressing a S14P/K97A mutant of hyoscyamine 6beta-hydroxylase AaH6H (2015), J. Biotechnol., 211, 123-129 .

Search for more literature for 1.14.11.11

Cloned(Commentary)

Cloned (Comment)	Organism
expression of wild-type and mutant enzymes in Escherichia coli	Anisodus acutangulus

Protein Variants

Protein Variants	Comment	Organism
K97A	the shows a 3.3fold improved hydroxylation activity compared with the wild-type enzyme	Anisodus acutangulus
K97E	the shows a 2.7fold improved hydroxylation activity compared with the wild-type enzyme	Anisodus acutangulus
K97G	the shows a 1.7fold improved hydroxylation activity compared with the wild-type enzyme	Anisodus acutangulus
S14H	the shows a 2.4fold improved hydroxylation activity compared with the wild-type enzyme	Anisodus acutangulus
S14P	the shows a 3.7fold improved hydroxylation activity compared with the wild-type enzyme	Anisodus acutangulus
S14P/K97A	double mutant shows a 4.4fold improved hydroxylation activity compared with the wild-type enzyme, and the in vivo epoxidation activity is also improved by 2.3times. With this mutant, in a 5 l bioreactor with a working volume of 3 l, scopolamine is produced via a single-enzyme-mediated two-step transformation from hyoscyamine of 500 mg/l in 97% yield (1.068 g)	Anisodus acutangulus

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
0.0348	-	L-hyoscyamine	pH 8.0, 35°C, mutant enzyme S14P/K97A	Anisodus acutangulus
0.0416	-	L-hyoscyamine	pH 8.0, 35°C	Anisodus acutangulus

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
L-hyoscyamine + 2-oxoglutarate + O2	Anisodus acutangulus	the enzyme catalyzes the hydroxylation of (-)-hyoscyamine and the subsequent epoxidation of 6beta-hydroxyhyoscyamine to form scopolamine, a natural alkaloid	(6S)-hydroxyhyoscyamine + succinate + CO2	-	?

Organism

Organism	UniProt	Comment	Textmining
Anisodus acutangulus	A2IBI0	-	-

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
L-hyoscyamine + 2-oxoglutarate + O2	-	Anisodus acutangulus	(6S)-hydroxyhyoscyamine + succinate + CO2	-	?
L-hyoscyamine + 2-oxoglutarate + O2	the enzyme catalyzes the hydroxylation of (-)-hyoscyamine and the subsequent epoxidation of 6beta-hydroxyhyoscyamine to form scopolamine, a natural alkaloid	Anisodus acutangulus	(6S)-hydroxyhyoscyamine + succinate + CO2	-	?

Synonyms

Synonyms	Comment	Organism
AaH6H	-	Anisodus acutangulus
hyoscyamine 6beta-hydroxylase	-	Anisodus acutangulus

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
35	-	assay at	Anisodus acutangulus

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism	Structure
1	-	L-hyoscyamine	pH 8.0, 35°C, wild-type enzyme	Anisodus acutangulus
3.9	-	L-hyoscyamine	pH 8.0, 35°C, mutant enzyme S14P/K97A	Anisodus acutangulus

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
8	-	assay at	Anisodus acutangulus

General Information

General Information	Comment	Organism
metabolism	the enzyme catalyzes the hydroxylation of (-)-hyoscyamine and the subsequent epoxidation of 6beta-hydroxyhyoscyamine to form scopolamine, a natural alkaloid	Anisodus acutangulus

kcat/KM [mM/s]

kcat/KM Value [1/mMs^-1]	kcat/KM Value Maximum [1/mMs^-1]	Substrate	Comment	Organism	Structure
24.04	-	L-hyoscyamine	pH 8.0, 35°C	Anisodus acutangulus
112.1	-	L-hyoscyamine	pH 8.0, 35°C, mutant enzyme S14P/K97A	Anisodus acutangulus

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Release 2023.1 (January 2023)