Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary for 1.1.1.30 extracted from

  • Nakashima, K.; Ito, K.; Nakajima, Y.; Yamazawa, R.; Miyakawa, S.; Yoshimoto, T.
    Closed complex of the D-3-hydroxybutyrate dehydrogenase induced by an enantiomeric competitive inhibitor (2009), J. Biochem., 145, 467-479.
    View publication on PubMed
Search for more literature for 1.1.1.30

Cloned(Commentary)

Cloned (Comment) Organism
wild-type and mutants expressed in Escherichia coli XL1Blue harbouring pHBDH11. Wild-type HBDH and mutants containing an N-terminal His-tag expressed in Escherichia coli XL1Blue using pQE30 as a vector Pseudomonas fragi

Crystallization (Commentary)

Crystallization (Comment) Organism
crystals of ternary complex of HBDH-NAD+-L-3-hydroxybutyrate and the binary complex of HBDH-NAD+. The former structure shows a closed-form conformation, which is considered an active form for catalysis, while the latter stays mostly in a open-form conformation. Crystals of mutants T190S and T190A Pseudomonas fragi

Protein Variants

Protein Variants Comment Organism
H144A catalytic efficiency (kcat/Km) is 0.2% of the activity of wild-type HBDH Pseudomonas fragi
K152A no activity Pseudomonas fragi
K152E very low activity Pseudomonas fragi
K152Q very low activity Pseudomonas fragi
K152R retains a significant level of activity Pseudomonas fragi
L215A both Km and kcat values are largely affected and the catalytic efficiency (kcat/Km) is less than 3% that of the wild-type enzyme Pseudomonas fragi
L215V Km values increase 3.5- and 4.3fold and the kcat values are 73-118% those of the wild-type toward D-3-hydroxybutyrate and acetoacetate, respectively. Mutation does not significantly change Km and kcat toward NAD+ and NADH Pseudomonas fragi
Q196A kcat/Km value is 0.6% that of the wild-type Pseudomonas fragi
Q196E substantially reduced activity Pseudomonas fragi
Q196N substantially reduced activity Pseudomonas fragi
Q94A catalytic efficiency (kcat/Km) is 1.4% of the activity of wild-type HBDH Pseudomonas fragi
T190A activity decreases to 0.1% that of the wild-type enzyme Pseudomonas fragi
T190C decreased activity Pseudomonas fragi
T190S retains 37% of the activity Pseudomonas fragi
W187A very low activity Pseudomonas fragi
W187F shows significant activity levels, 65% that of the wild-type enzyme Pseudomonas fragi
W187T shows faint activity Pseudomonas fragi
W187Y shows significant activity levels, 41% that of the wild-type enzyme Pseudomonas fragi
W257A no activity Pseudomonas fragi
W257F shows low activity levels, 2% that of the wild-type enzyme Pseudomonas fragi
W257Y shows low activity levels, 1% that of the wild-type enzyme Pseudomonas fragi
Y155F no activity Pseudomonas fragi

Inhibitors

Inhibitors Comment Organism Structure
L-3-hydroxybutyrate is a competitive inhibitor Pseudomonas fragi

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.01
-
 NADH wild-type Pseudomonas fragi
0.02
-
 NADH mutant T190S Pseudomonas fragi
0.03
-
 NADH mutant L215V Pseudomonas fragi
0.1
-
 NADH mutant T190A Pseudomonas fragi
0.12
-
 NAD+ mutant H6-K152R Pseudomonas fragi
0.12
-
 NAD+ mutant H6-Q196A Pseudomonas fragi
0.12
-
 NAD+ mutant H6-Q196E Pseudomonas fragi
0.17
-
 NAD+ H6-HBDH Pseudomonas fragi
0.19
-
 NAD+ mutant H6-Q94A Pseudomonas fragi
0.2
-
 NAD+ mutant H6-Q196N Pseudomonas fragi
0.21
-
 NAD+ mutant H6-W187Y Pseudomonas fragi
0.24
-
 NAD+ wild-type Pseudomonas fragi
0.24
-
 NAD+ mutant H6-W187F Pseudomonas fragi
0.29
-
 NAD+ mutant L215V Pseudomonas fragi
0.35
-
 NAD+ mutant H6-H144A Pseudomonas fragi
0.37
-
 acetoacetate wild-type Pseudomonas fragi
0.49
-
 NAD+ mutant H6-W257F Pseudomonas fragi
0.5
-
 NADH mutant L215A Pseudomonas fragi
0.69
-
 NAD+ mutant T190C Pseudomonas fragi
0.71
-
 NAD+ mutant T190S Pseudomonas fragi
0.74
-
 NAD+ mutant H6-W257Y Pseudomonas fragi
0.76
-
 NAD+ mutant H6-W187A Pseudomonas fragi
0.8
-
 D-3-hydroxybutyrate wild-type Pseudomonas fragi
0.88
-
 D-3-hydroxybutyrate H6-HBDH Pseudomonas fragi
0.88
-
 NAD+ mutant H6-W187T Pseudomonas fragi
0.95
-
 NAD+ mutant H6-W257A Pseudomonas fragi
1.1
-
 acetoacetate mutant T190S Pseudomonas fragi
1.2
-
 D-3-hydroxybutyrate mutant H6-W187Y Pseudomonas fragi
1.3
-
 D-3-hydroxybutyrate mutant H6-W187F Pseudomonas fragi
1.3
-
 acetoacetate mutant L215V Pseudomonas fragi
1.5
-
 NAD+ mutant L215A Pseudomonas fragi
1.5
-
 NADH mutant T190C Pseudomonas fragi
2.2
-
 NAD+ mutant T190A Pseudomonas fragi
3.4
-
 D-3-hydroxybutyrate mutant L215V Pseudomonas fragi
3.7
-
 D-3-hydroxybutyrate mutant T190S Pseudomonas fragi
7.7
-
 acetoacetate mutant T190C Pseudomonas fragi
8.3
-
 acetoacetate mutant L215A Pseudomonas fragi
9.2
-
 D-3-hydroxybutyrate mutant L215A Pseudomonas fragi
25
-
 D-3-hydroxybutyrate mutant H6-H144A Pseudomonas fragi
25
-
 D-3-hydroxybutyrate mutant T190A Pseudomonas fragi
30
-
 D-3-hydroxybutyrate mutant H6-Q94A Pseudomonas fragi
31
-
 D-3-hydroxybutyrate mutant H6-K152R Pseudomonas fragi
32
-
 acetoacetate mutant T190A Pseudomonas fragi
36
-
 D-3-hydroxybutyrate mutant T190C Pseudomonas fragi
47
-
 D-3-hydroxybutyrate mutant H6-Q196A Pseudomonas fragi
51
-
 D-3-hydroxybutyrate mutant H6-Q196N Pseudomonas fragi
61
-
 D-3-hydroxybutyrate mutant H6-W257A Pseudomonas fragi
65
-
 D-3-hydroxybutyrate mutant H6-W257Y Pseudomonas fragi
83
-
 D-3-hydroxybutyrate mutant H6-Q196E Pseudomonas fragi
83
-
 D-3-hydroxybutyrate mutant H6-W257F Pseudomonas fragi
84
-
 D-3-hydroxybutyrate mutant H6-W187T Pseudomonas fragi
100
-
 D-3-hydroxybutyrate mutant H6-W187A Pseudomonas fragi

Organism

Organism UniProt Comment Textmining
Pseudomonas fragi Q5KST5
-
-

Purification (Commentary)

Purification (Comment) Organism
wild-type and mutants purified by metal chelating affinity chromatography followed by ion-exchange chromatography Pseudomonas fragi

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
D-3-hydroxybutyrate + NAD+
-
 Pseudomonas fragi acetoacetate + NADH + H+
-
 r

Synonyms

Synonyms Comment Organism
HBDH
-
 Pseudomonas fragi

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
4.5
-
 acetoacetate mutant T190A Pseudomonas fragi
5.1
-
 acetoacetate mutant T190C Pseudomonas fragi
6.1
-
 D-3-hydroxybutyrate mutant T190C Pseudomonas fragi
6.7
-
 D-3-hydroxybutyrate mutant H6-W257A Pseudomonas fragi
7.2
-
 D-3-hydroxybutyrate mutant H6-W187T Pseudomonas fragi
12
-
 D-3-hydroxybutyrate mutant H6-W257Y Pseudomonas fragi
21
-
 D-3-hydroxybutyrate mutant H6-H144A Pseudomonas fragi
22
-
 D-3-hydroxybutyrate mutant H6-Q196N Pseudomonas fragi
22
-
 D-3-hydroxybutyrate mutant T190A Pseudomonas fragi
29
-
 D-3-hydroxybutyrate mutant H6-W187A Pseudomonas fragi
30
-
 acetoacetate mutant L215A Pseudomonas fragi
70
-
 D-3-hydroxybutyrate mutant L215A Pseudomonas fragi
83
-
 D-3-hydroxybutyrate mutant H6-Q196E Pseudomonas fragi
87
-
 acetoacetate mutant L215V Pseudomonas fragi
88
-
 D-3-hydroxybutyrate mutant H6-W257F Pseudomonas fragi
99
-
 D-3-hydroxybutyrate mutant H6-Q196A Pseudomonas fragi
110
-
 D-3-hydroxybutyrate mutant H6-K152R Pseudomonas fragi
120
-
 acetoacetate wild-type Pseudomonas fragi
125
-
 acetoacetate mutant T190S Pseudomonas fragi
140
-
 D-3-hydroxybutyrate mutant H6-Q94A Pseudomonas fragi
180
-
 D-3-hydroxybutyrate mutant H6-W187Y Pseudomonas fragi
310
-
 D-3-hydroxybutyrate H6-HBDH Pseudomonas fragi
310
-
 D-3-hydroxybutyrate mutant H6-W187F Pseudomonas fragi
370
-
 D-3-hydroxybutyrate wild-type Pseudomonas fragi
435
-
 D-3-hydroxybutyrate mutant L215V Pseudomonas fragi
705
-
 D-3-hydroxybutyrate mutant T190S Pseudomonas fragi

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8.5
-
-
 Pseudomonas fragi