Literature summary for 1.4.1.16 extracted from

Gao, X.; Huang, F.; Feng, J.; Chen, X.; Zhang, H.; Wang, Z.; Wu, Q.; Zhu, D.
Engineering the meso-diaminopimelate dehydrogenase from Symbiobacterium thermophilum by site saturation mutagenesis for D-phenylalanine synthesis (2013), Appl. Environ. Microbiol., 79, 5078-5081.

Search for more literature for 1.4.1.16

Cloned(Commentary)

Cloned (Comment)	Organism
recombinant expression of wild-type and mutant enzymes in Escherichia coli strain BL21(DE3)	Symbiobacterium thermophilum

Protein Variants

Protein Variants	Comment	Organism
H227C	site-directed saturation mutagenesis, the mutant shows 15.1fold increased activity with phenylpyruvate compared to the wild-type enzyme	Symbiobacterium thermophilum
H227V	site-directed saturation mutagenesis, the mutant shows 35.1fold increased activity with phenylpyruvate compared to the wild-type enzyme	Symbiobacterium thermophilum
additional information	in order to enlarge the substrate binding pocket of the meso-diaminopimelate dehydrogenase from Symbiobacterium thermophilum to accommodate larger 2-keto acids, e.g. phenylalanine, four amino acid residues, Phe146, Thr171, Arg181, and His227, are targeted for site saturation mutagenesis	Symbiobacterium thermophilum
R181F	site-directed saturation mutagenesis, the mutant shows 6.4fold increased activity with phenylpyruvate compared to the wild-type enzyme	Symbiobacterium thermophilum
R181F/H227V	site-directed saturation mutagenesis, the mutant shows 19.3fold increased activity with phenylpyruvate compared to the wild-type enzyme	Symbiobacterium thermophilum
T171P	site-directed saturation mutagenesis, the mutant shows 2.2fold increased activity with phenylpyruvate compared to the wild-type enzyme	Symbiobacterium thermophilum
T171S	site-directed saturation mutagenesis, the mutant shows 2.8fold increased activity with phenylpyruvate compared to the wild-type enzyme	Symbiobacterium thermophilum
T171S/H227V	site-directed saturation mutagenesis, the mutant shows 10.6fold increased activity with phenylpyruvate compared to the wild-type enzyme	Symbiobacterium thermophilum
T171S/R181F	site-directed saturation mutagenesis, the mutant shows 4.0fold increased activity with phenylpyruvate compared to the wild-type enzyme	Symbiobacterium thermophilum
T171S/R181F/H227V	site-directed saturation mutagenesis, the mutant shows 14.6fold increased activity with phenylpyruvate compared to the wild-type enzyme	Symbiobacterium thermophilum

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
11	-	phenylpyruvate	pH 8.5, 30°C, recombinant mutant T171S/R181F/H227V	Symbiobacterium thermophilum
11.1	-	phenylpyruvate	pH 8.5, 30°C, recombinant mutant R181F	Symbiobacterium thermophilum
12.5	-	phenylpyruvate	pH 8.5, 30°C, recombinant wild-type enzyme	Symbiobacterium thermophilum
13.9	-	phenylpyruvate	pH 8.5, 30°C, recombinant mutant T171S/H227V	Symbiobacterium thermophilum
13.9	-	phenylpyruvate	pH 8.5, 30°C, recombinant mutant T171S/R181F	Symbiobacterium thermophilum
15.5	-	phenylpyruvate	pH 8.5, 30°C, recombinant mutant H227C	Symbiobacterium thermophilum
15.8	-	phenylpyruvate	pH 8.5, 30°C, recombinant mutantT171P	Symbiobacterium thermophilum
19.6	-	phenylpyruvate	pH 8.5, 30°C, recombinant mutant T171S	Symbiobacterium thermophilum
20.8	-	phenylpyruvate	pH 8.5, 30°C, recombinant mutant R181F/H227V	Symbiobacterium thermophilum
24.3	-	phenylpyruvate	pH 8.5, 30°C, recombinant mutant H227V	Symbiobacterium thermophilum

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
meso-2,6-diaminoheptanedioate + H2O + NADP+	Symbiobacterium thermophilum	-	L-2-amino-6-oxoheptanedioate + NH3 + NADPH + H+	-	r

Organism

Organism	UniProt	Comment	Textmining
Symbiobacterium thermophilum	Q67PI3	-	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant wild-type and mutant enzymes from Escherichia coli strain Bl21(DE3)	Symbiobacterium thermophilum

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg]	Specific Activity Maximum [µmol/min/mg]	Comment	Organism
0.07	-	recombinant wild-type enzyme, pH 8.5, 30°C	Symbiobacterium thermophilum
0.15	-	recombinant mutant T171P, pH 8.5, 30°C	Symbiobacterium thermophilum
0.19	-	recombinant mutant T171S, pH 8.5, 30°C	Symbiobacterium thermophilum
0.28	-	recombinant mutant T171S/R181F, pH 8.5, 30°C	Symbiobacterium thermophilum
0.44	-	recombinant mutant R181F, pH 8.5, 30°C	Symbiobacterium thermophilum
0.74	-	recombinant mutant T171S/H227V, pH 8.5, 30°C	Symbiobacterium thermophilum
1.02	-	recombinant mutant T171S/R181F/H227V, pH 8.5, 30°C	Symbiobacterium thermophilum
1.03	-	recombinant mutant H227C, pH 8.5, 30°C	Symbiobacterium thermophilum
1.35	-	recombinant mutant R181F/H227V, pH 8.5, 30°C	Symbiobacterium thermophilum
2.39	-	recombinant mutant H227V, pH 8.5, 30°C	Symbiobacterium thermophilum

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
meso-2,6-diaminoheptanedioate + H2O + NADP+	-	Symbiobacterium thermophilum	L-2-amino-6-oxoheptanedioate + NH3 + NADPH + H+	-	r
phenylpyruvate + NH3 + NADPH + H+	-	Symbiobacterium thermophilum	phenylalanine + NADP+	-	?

Synonyms

Synonyms	Comment	Organism
meso-DAPDH	-	Symbiobacterium thermophilum
meso-diaminopimelate dehydrogenase	-	Symbiobacterium thermophilum

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
30	-	assay at	Symbiobacterium thermophilum

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism	Structure
0.11	-	phenylpyruvate	pH 8.5, 30°C, recombinant wild-type enzyme	Symbiobacterium thermophilum
0.53	-	phenylpyruvate	pH 8.5, 30°C, recombinant mutantT171P	Symbiobacterium thermophilum
0.7	-	phenylpyruvate	pH 8.5, 30°C, recombinant mutant T171S/R181F	Symbiobacterium thermophilum
0.88	-	phenylpyruvate	pH 8.5, 30°C, recombinant mutant T171S	Symbiobacterium thermophilum
1.07	-	phenylpyruvate	pH 8.5, 30°C, recombinant mutant R181F	Symbiobacterium thermophilum
2.09	-	phenylpyruvate	pH 8.5, 30°C, recombinant mutant T171S/R181F/H227V	Symbiobacterium thermophilum
2.37	-	phenylpyruvate	pH 8.5, 30°C, recombinant mutant T171S/H227V	Symbiobacterium thermophilum
2.48	-	phenylpyruvate	pH 8.5, 30°C, recombinant mutant H227C	Symbiobacterium thermophilum
3.43	-	phenylpyruvate	pH 8.5, 30°C, recombinant mutant R181F/H227V	Symbiobacterium thermophilum
3.98	-	phenylpyruvate	pH 8.5, 30°C, recombinant mutant H227V	Symbiobacterium thermophilum

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
8.5	-	assay at	Symbiobacterium thermophilum

Cofactor

Cofactor	Comment	Organism	Structure
NADP+	-	Symbiobacterium thermophilum
NADPH	-	Symbiobacterium thermophilum

General Information

General Information	Comment	Organism
additional information	molecular dynamic simulation analysis of wild-type and variant H227V enzyme with the imine intermediate as the substrate, overview	Symbiobacterium thermophilum

Use of this online version of BRENDA is free under the CC BY 4.0 license. See terms of use for full details.

Release 2023.1 (January 2023)