Literature summary for 3.5.2.B2 extracted from

Gao, S.; Zhu, S.; Huang, R.; Li, H.; Wang, H.; Zheng, G.
Engineering the enantioselectivity and thermostability of a (+)-?-lactamase from Microbacterium hydrocarbonoxydans for kinetic resolution of Vince lactam (2-azabicyclo[2.2.1]hept-5-en-3-one) (2018), Appl. Environ. Microbiol., 84, e01780-17 .

Search for more literature for 3.5.2.B2

Protein Variants

Protein Variants	Comment	Organism
E95K/V54S	optimal temperature is 10°C higher compared to wild-type enzyme. Mutant enzyme may be suitable for preparation of the optically pure intermediate (-)-Vince lactam.	Microbacterium hydrocarbonoxydans
F110A/V112G	mutant enzyme shows no activity with (-)-Vince lactam	Microbacterium hydrocarbonoxydans
H51A	mutant enzyme shows no activity with (-)-Vince lactam	Microbacterium hydrocarbonoxydans
R162T/V54L	optimal temperature is 15°C higher compared to wild-type enzyme	Microbacterium hydrocarbonoxydans
V112A	mutant enzyme shows no activity with (-)-Vince lactam	Microbacterium hydrocarbonoxydans
V54L	mutant enzyme shows no activity with (-)-Vince lactam	Microbacterium hydrocarbonoxydans
V54S	mutant enzyme shows no activity with (-)-Vince lactam	Microbacterium hydrocarbonoxydans
V54T	mutant enzyme shows no activity with (-)-Vince lactam	Microbacterium hydrocarbonoxydans

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
12	-	2-azabicyclo[2.2.1]hept-5-en-3-one	pH 7.5, 30°C, wild-type enzyme	Microbacterium hydrocarbonoxydans
18	-	2-azabicyclo[2.2.1]hept-5-en-3-one	pH 7.5, 30°C, mutant enzyme E95V/V54S	Microbacterium hydrocarbonoxydans
18	-	2-azabicyclo[2.2.1]hept-5-en-3-one	pH 7.5, 30°C, mutant enzyme R162T/V54S	Microbacterium hydrocarbonoxydans
18	-	2-azabicyclo[2.2.1]hept-5-en-3-one	pH 7.5, 30°C, mutant enzyme V54S	Microbacterium hydrocarbonoxydans
20	-	2-azabicyclo[2.2.1]hept-5-en-3-one	pH 7.5, 30°C, mutant enzyme E95K/V54S	Microbacterium hydrocarbonoxydans
20.2	-	2-azabicyclo[2.2.1]hept-5-en-3-one	pH 7.5, 30°C, mutant enzyme E95K/V54L	Microbacterium hydrocarbonoxydans
20.2	-	2-azabicyclo[2.2.1]hept-5-en-3-one	pH 7.5, 30°C, mutant enzyme R162T/V54L	Microbacterium hydrocarbonoxydans
24	-	2-azabicyclo[2.2.1]hept-5-en-3-one	pH 7.5, 30°C, mutant enzyme E95Q/V54S	Microbacterium hydrocarbonoxydans
26.3	-	2-azabicyclo[2.2.1]hept-5-en-3-one	pH 7.5, 30°C, mutant enzyme E95V/V54L	Microbacterium hydrocarbonoxydans
28.1	-	2-azabicyclo[2.2.1]hept-5-en-3-one	pH 7.5, 30°C, mutant enzyme E95Q/V54L	Microbacterium hydrocarbonoxydans
29	-	2-azabicyclo[2.2.1]hept-5-en-3-one	pH 7.5, 30°C, mutant enzyme V54L	Microbacterium hydrocarbonoxydans
29	-	2-azabicyclo[2.2.1]hept-5-en-3-one	pH 7.5, 30°C, mutant enzyme V54T	Microbacterium hydrocarbonoxydans
38	-	2-azabicyclo[2.2.1]hept-5-en-3-one	pH 7.5, 30°C, mutant enzyme V112A	Microbacterium hydrocarbonoxydans
39	-	2-azabicyclo[2.2.1]hept-5-en-3-one	pH 7.5, 30°C, mutant enzyme F110A/V112G	Microbacterium hydrocarbonoxydans
40	-	2-azabicyclo[2.2.1]hept-5-en-3-one	pH 7.5, 30°C, mutant enzyme H51A	Microbacterium hydrocarbonoxydans

Organism

Organism	UniProt	Comment	Textmining
Microbacterium hydrocarbonoxydans	A0A0K0XHU0	-	-

Purification (Commentary)

Purification (Comment)	Organism
-	Microbacterium hydrocarbonoxydans

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
2-azabicyclo[2.2.1]hept-5-en-3-one + H2O	i.e. Vince lactam. Wild-type enzyme shows activity on (+)-Vince lactam and (-)-Vince lactam. The mutant enzymes V54S, V54L, V112A, H51A, F110A/V112G and V54T show no activity with (-)-Vince lactam	Microbacterium hydrocarbonoxydans	(-)-2-azabicyclo[2.2.1]hept-5-en-3-one + (+)-4-amino-cyclopent-2-enecarboxylic acid	-	?

Synonyms

Synonyms	Comment	Organism
MhIHL	-	Microbacterium hydrocarbonoxydans

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
25	-	wild-type enzyme	Microbacterium hydrocarbonoxydans
30	-	assay at	Microbacterium hydrocarbonoxydans
35	-	mutant enzyme E95K/V54S	Microbacterium hydrocarbonoxydans
40	-	mutant enzyme R162T/V54L	Microbacterium hydrocarbonoxydans

Temperature Range [°C]

Temperature Minimum [°C]	Temperature Maximum [°C]	Comment	Organism
20	55	20°C: about 70% of maximal activity, 55°C: about 55% of maximal activity, mutant enzyme E95K/V54S	Microbacterium hydrocarbonoxydans
20	55	20°C: about 75% of maximal activity, 55°C: about 60% of maximal activity, mutant enzyme R162T/V54L	Microbacterium hydrocarbonoxydans
20	45	20°C: about 80% of maximal activity, 45°C: about 40% of maximal activity, wild-type enzyme	Microbacterium hydrocarbonoxydans

Temperature Stability [°C]

Temperature Stability Minimum [°C]	Temperature Stability Maximum [°C]	Comment	Organism
41	-	15 min, 50% loss of activity, wild-type enzyme	Microbacterium hydrocarbonoxydans
48	-	15 min, 50% loss of activity, mutant enzyme G42T/V56M/G57V	Microbacterium hydrocarbonoxydans
57	-	15 min, 50% loss of activity, mutant enzyme R162T	Microbacterium hydrocarbonoxydans
58	-	15 min, 50% loss of activity, mutant enzyme E95V	Microbacterium hydrocarbonoxydans
60	-	15 min, 50% loss of activity, mutant enzyme E95F	Microbacterium hydrocarbonoxydans
61	-	15 min, 50% loss of activity, mutant enzyme E95V/V54L	Microbacterium hydrocarbonoxydans
62	-	15 min, 50% loss of activity, mutant enzyme E95V/V54S	Microbacterium hydrocarbonoxydans
65	-	15 min, 50% loss of activity, mutant enzyme E95Q/V54L	Microbacterium hydrocarbonoxydans
66	-	15 min, 50% loss of activity, mutant enzyme R162T/V54S	Microbacterium hydrocarbonoxydans
67	-	15 min, 50% loss of activity, mutant enzyme E95Q/V54S	Microbacterium hydrocarbonoxydans
68	-	15 min, 50% loss of activity, mutant enzyme E95K/V54L	Microbacterium hydrocarbonoxydans
69	-	15 min, 50% loss of activity, mutant enzyme R162T/V54L	Microbacterium hydrocarbonoxydans
70	-	15 min, 50% loss of activity, mutant enzyme E95K/V54S	Microbacterium hydrocarbonoxydans
70	-	15 min, 50% loss of activity, mutant enzyme E95Q	Microbacterium hydrocarbonoxydans
72	-	15 min, 50% loss of activity, mutant enzyme E95K	Microbacterium hydrocarbonoxydans

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism	Structure
7.8	-	2-azabicyclo[2.2.1]hept-5-en-3-one	pH 7.5, 30°C, mutant enzyme F110A/V112G	Microbacterium hydrocarbonoxydans
11	-	2-azabicyclo[2.2.1]hept-5-en-3-one	pH 7.5, 30°C, mutant enzyme V112A	Microbacterium hydrocarbonoxydans
49	-	2-azabicyclo[2.2.1]hept-5-en-3-one	pH 7.5, 30°C, mutant enzyme V54T	Microbacterium hydrocarbonoxydans
72	-	2-azabicyclo[2.2.1]hept-5-en-3-one	pH 7.5, 30°C, mutant enzyme H51A	Microbacterium hydrocarbonoxydans
101	-	2-azabicyclo[2.2.1]hept-5-en-3-one	pH 7.5, 30°C, mutant enzyme E95Q/V54L	Microbacterium hydrocarbonoxydans
120	-	2-azabicyclo[2.2.1]hept-5-en-3-one	pH 7.5, 30°C, mutant enzyme E95V/V54S	Microbacterium hydrocarbonoxydans
123	-	2-azabicyclo[2.2.1]hept-5-en-3-one	pH 7.5, 30°C, mutant enzyme E95K/V54L	Microbacterium hydrocarbonoxydans
130	-	2-azabicyclo[2.2.1]hept-5-en-3-one	pH 7.5, 30°C, mutant enzyme R162T/V54S	Microbacterium hydrocarbonoxydans
140	-	2-azabicyclo[2.2.1]hept-5-en-3-one	pH 7.5, 30°C, mutant enzyme V54S	Microbacterium hydrocarbonoxydans
143	-	2-azabicyclo[2.2.1]hept-5-en-3-one	pH 7.5, 30°C, mutant enzyme E95Q/V54S	Microbacterium hydrocarbonoxydans
159	-	2-azabicyclo[2.2.1]hept-5-en-3-one	pH 7.5, 30°C, mutant enzyme R162T/V54L	Microbacterium hydrocarbonoxydans
164	-	2-azabicyclo[2.2.1]hept-5-en-3-one	pH 7.5, 30°C, mutant enzyme E95K/V54S	Microbacterium hydrocarbonoxydans
168	-	2-azabicyclo[2.2.1]hept-5-en-3-one	pH 7.5, 30°C, mutant enzyme E95V/V54L	Microbacterium hydrocarbonoxydans
190	-	2-azabicyclo[2.2.1]hept-5-en-3-one	pH 7.5, 30°C, mutant enzyme V54L	Microbacterium hydrocarbonoxydans
200	-	2-azabicyclo[2.2.1]hept-5-en-3-one	pH 7.5, 30°C, wild-type enzyme	Microbacterium hydrocarbonoxydans

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
7.5	-	assay at	Microbacterium hydrocarbonoxydans

kcat/KM [mM/s]

kcat/KM Value [1/mMs^-1]	kcat/KM Value Maximum [1/mMs^-1]	Substrate	Comment	Organism	Structure
0.2	-	2-azabicyclo[2.2.1]hept-5-en-3-one	pH 7.5, 30°C, mutant enzyme F110A/V112G	Microbacterium hydrocarbonoxydans
0.3	-	2-azabicyclo[2.2.1]hept-5-en-3-one	pH 7.5, 30°C, mutant enzyme V112A	Microbacterium hydrocarbonoxydans
1.7	-	2-azabicyclo[2.2.1]hept-5-en-3-one	pH 7.5, 30°C, mutant enzyme V54T	Microbacterium hydrocarbonoxydans
1.8	-	2-azabicyclo[2.2.1]hept-5-en-3-one	pH 7.5, 30°C, mutant enzyme H51A	Microbacterium hydrocarbonoxydans
3.6	-	2-azabicyclo[2.2.1]hept-5-en-3-one	pH 7.5, 30°C, mutant enzyme E95Q/V54L	Microbacterium hydrocarbonoxydans
5.9	-	2-azabicyclo[2.2.1]hept-5-en-3-one	pH 7.5, 30°C, mutant enzyme E95Q/V54S	Microbacterium hydrocarbonoxydans
6.1	-	2-azabicyclo[2.2.1]hept-5-en-3-one	pH 7.5, 30°C, mutant enzyme E95K/V54L	Microbacterium hydrocarbonoxydans
6.4	-	2-azabicyclo[2.2.1]hept-5-en-3-one	pH 7.5, 30°C, mutant enzyme E95V/V54L	Microbacterium hydrocarbonoxydans
6.5	-	2-azabicyclo[2.2.1]hept-5-en-3-one	pH 7.5, 30°C, mutant enzyme E95V/V54S	Microbacterium hydrocarbonoxydans
6.7	-	2-azabicyclo[2.2.1]hept-5-en-3-one	pH 7.5, 30°C, mutant enzyme V54L	Microbacterium hydrocarbonoxydans
7.2	-	2-azabicyclo[2.2.1]hept-5-en-3-one	pH 7.5, 30°C, mutant enzyme R162T/V54S	Microbacterium hydrocarbonoxydans
7.9	-	2-azabicyclo[2.2.1]hept-5-en-3-one	pH 7.5, 30°C, mutant enzyme R162T/V54L	Microbacterium hydrocarbonoxydans
8.2	-	2-azabicyclo[2.2.1]hept-5-en-3-one	pH 7.5, 30°C, mutant enzyme E95K/V54S	Microbacterium hydrocarbonoxydans
8.2	-	2-azabicyclo[2.2.1]hept-5-en-3-one	pH 7.5, 30°C, mutant enzyme V54S	Microbacterium hydrocarbonoxydans
15.77	-	2-azabicyclo[2.2.1]hept-5-en-3-one	pH 7.5, 30°C, wild-type enzyme	Microbacterium hydrocarbonoxydans

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Release 2023.1 (January 2023)