Literature summary for 1.4.1.20 extracted from

Chen, S.; Engel, P.C.
Efficient screening for new amino acid dehydrogenase activity: directed evolution of Bacillus sphaericus phenylalanine dehydrogenase towards activity with an unsaturated non-natural amino acid (2009), J. Biotechnol., 142, 127-134.

Cloned(Commentary)

Cloned (Comment)	Organism
expressed in Escherichia coli via plasmids	Lysinibacillus sphaericus

Protein Variants

Protein Variants	Comment	Organism
D126N	mutant without enzymatic activity	Lysinibacillus sphaericus
D209G	mutant named 1stA7, active with DL-propargylglycine	Lysinibacillus sphaericus
D209G/Q18H/I336F	mutant named 1stA7/1stB6, active with DL-propargylglycine	Lysinibacillus sphaericus
E313G	mutant named H7H10, active with DL-propargylglycine	Lysinibacillus sphaericus
F110Y/G124C/G293A	mutant named H14A12, active with DL-propargylglycine	Lysinibacillus sphaericus
G124A	mutant named 9F5, active with DL-propargylglycine	Lysinibacillus sphaericus
G124A/E313G	mutant named 25B12, active with DL-propargylglycine	Lysinibacillus sphaericus
G124A/E313N	mutant named 20E1, active with DL-propargylglycine	Lysinibacillus sphaericus
K224Q/L283F	mutant named 2ndB10, active with DL-propargylglycine	Lysinibacillus sphaericus
K224Q/L283F/E313G	mutant named 2ndB10/H7H10, active with DL-propargylglycine	Lysinibacillus sphaericus
Q18H/I336F	mutant named 1stB6, active with DL-propargylglycine	Lysinibacillus sphaericus
Q18H/I336F/E313G	mutant named 1stB6/H7H10, active with DL-propargylglycine	Lysinibacillus sphaericus
V135I/I308N/Q363H	mutant named H21D1, active with DL-propargylglycine	Lysinibacillus sphaericus
V33A/A206D/L283F	mutant named 2ndC2., active with DL-propargylglycine	Lysinibacillus sphaericus

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism
0.018	-	L-phenylalanine	mutations Q18H/I336F/E313G	Lysinibacillus sphaericus
0.02	-	L-phenylalanine	mutations K224Q/L283F/E313G	Lysinibacillus sphaericus
0.035	-	L-phenylalanine	mutations V135I/I308N/Q363H	Lysinibacillus sphaericus
0.04	-	L-phenylalanine	mutations K224Q/L283F	Lysinibacillus sphaericus
0.06	-	L-phenylalanine	mutations V33A/A206D/L283F	Lysinibacillus sphaericus
0.07	-	L-phenylalanine	mutations D209G/Q18H/I336F	Lysinibacillus sphaericus
0.07	-	L-phenylalanine	mutations Q18H/I336F	Lysinibacillus sphaericus
0.12	-	L-phenylalanine	mutation E313G	Lysinibacillus sphaericus
0.67	-	L-phenylalanine	mutation D209G	Lysinibacillus sphaericus
1.4	-	L-phenylalanine	wild type enzyme of Bacillus sphaericus	Lysinibacillus sphaericus
1.51	-	L-phenylalanine	mutations F110Y/G124C/G293A	Lysinibacillus sphaericus
2	3	DL-propargylglycine	mutations G124A/E313G	Lysinibacillus sphaericus
4.5	-	L-phenylalanine	mutations G124A/E313N	Lysinibacillus sphaericus
7.9	-	L-phenylalanine	mutation G124A	Lysinibacillus sphaericus
9.6	-	L-phenylalanine	mutations G124A/E313G	Lysinibacillus sphaericus
28	-	DL-propargylglycine	mutation E313G	Lysinibacillus sphaericus
28	-	DL-propargylglycine	mutation G124A	Lysinibacillus sphaericus
32	-	DL-propargylglycine	mutations K224Q/L283F/E313G	Lysinibacillus sphaericus
32	-	DL-propargylglycine	mutations Q18H/I336F	Lysinibacillus sphaericus
34	-	DL-propargylglycine	mutations K224Q/L283F	Lysinibacillus sphaericus
39	-	DL-propargylglycine	mutations Q18H/I336F/E313G	Lysinibacillus sphaericus
42	-	DL-propargylglycine	mutations F110Y/G124C/G293A	Lysinibacillus sphaericus
44	-	DL-propargylglycine	mutations V33A/A206D/L283F	Lysinibacillus sphaericus
48	-	DL-propargylglycine	mutations G124A/E313N	Lysinibacillus sphaericus
58	-	DL-propargylglycine	mutations D209G/Q18H/I336F	Lysinibacillus sphaericus
84	-	DL-propargylglycine	mutations V135I/I308N/Q363H	Lysinibacillus sphaericus
134	-	DL-propargylglycine	wild type enzyme of Bacillus sphaericus	Lysinibacillus sphaericus
298	-	DL-propargylglycine	mutation D209G	Lysinibacillus sphaericus

Organism

Organism	UniProt	Comment	Textmining
Lysinibacillus sphaericus	-	-	-

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
DL-propargylglycine + phenazine ethosulfate + 2-(4-iodophenyl)-3-(4-nitrophenyl)-5-phenyltetrazolium chloridehydrate + NAD+	substrate is poorly fertilized by wild type enzyme, but using muteins with different mutations higher turnover can be achieved	Lysinibacillus sphaericus	? + NADH	-	?
L-phenylalanine + H2O + NAD+	-	Lysinibacillus sphaericus	phenylpyruvate + NH3 + NADH + H+	-	?

Synonyms

Synonyms	Comment	Organism
PheDH	-	Lysinibacillus sphaericus
phenylalanine dehydrogenase	-	Lysinibacillus sphaericus

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism
0.5	-	DL-propargylglycine	mutations K224Q/L283F/E313G	Lysinibacillus sphaericus
0.57	-	DL-propargylglycine	mutations G124A/E313N	Lysinibacillus sphaericus
0.59	-	DL-propargylglycine	mutations Q18H/I336F/E313G	Lysinibacillus sphaericus
0.608	-	L-phenylalanine	mutations V135I/I308N/Q363H	Lysinibacillus sphaericus
0.69	-	DL-propargylglycine	mutation G124A	Lysinibacillus sphaericus
0.722	-	DL-propargylglycine	wild type enzyme of Bacillus sphaericus	Lysinibacillus sphaericus
0.759	-	DL-propargylglycine	mutations F110Y/G124C/G293A	Lysinibacillus sphaericus
0.91	-	L-phenylalanine	mutations Q18H/I336F/E313G	Lysinibacillus sphaericus
0.92	-	DL-propargylglycine	mutations G124A/E313G	Lysinibacillus sphaericus
1.02	-	DL-propargylglycine	mutations Q18H/I336F	Lysinibacillus sphaericus
1.07	-	DL-propargylglycine	mutation E313G	Lysinibacillus sphaericus
1.09	-	L-phenylalanine	mutations K224Q/L283F/E313G	Lysinibacillus sphaericus
1.12	-	L-phenylalanine	mutations F110Y/G124C/G293A	Lysinibacillus sphaericus
1.15	-	DL-propargylglycine	mutations K224Q/L283F	Lysinibacillus sphaericus
1.3	-	DL-propargylglycine	mutations D209G/Q18H/I336F	Lysinibacillus sphaericus
1.42	-	DL-propargylglycine	mutations V33A/A206D/L283F	Lysinibacillus sphaericus
1.49	-	DL-propargylglycine	mutations V135I/I308N/Q363H	Lysinibacillus sphaericus
1.82	-	DL-propargylglycine	mutation D209G	Lysinibacillus sphaericus
2.6	-	L-phenylalanine	mutation G124A	Lysinibacillus sphaericus
3.07	-	L-phenylalanine	mutations G124A/E313N	Lysinibacillus sphaericus
3.16	-	L-phenylalanine	mutations K224Q/L283F	Lysinibacillus sphaericus
4.08	-	L-phenylalanine	mutations D209G/Q18H/I336F	Lysinibacillus sphaericus
4.66	-	L-phenylalanine	mutations G124A/E313G	Lysinibacillus sphaericus
5.8	-	L-phenylalanine	mutations V33A/A206D/L283F	Lysinibacillus sphaericus
6.56	-	L-phenylalanine	mutations Q18H/I336F	Lysinibacillus sphaericus
32.1	-	L-phenylalanine	mutation E313G	Lysinibacillus sphaericus
56.1	-	L-phenylalanine	mutation D209G	Lysinibacillus sphaericus
57.1	-	L-phenylalanine	wild type enzyme of Bacillus sphaericus	Lysinibacillus sphaericus

kcat/KM [mM/s]

kcat/KM Value [1/mMs^-1]	kcat/KM Value Maximum [1/mMs^-1]	Substrate	Comment	Organism
0.0054	-	DL-propargylglycine	wild type enzyme of Bacillus sphaericus	Lysinibacillus sphaericus
0.0061	-	DL-propargylglycine	mutation D209G	Lysinibacillus sphaericus
0.012	-	DL-propargylglycine	mutations G124A/E313N	Lysinibacillus sphaericus
0.015	-	DL-propargylglycine	mutations K224Q/L283F/E313G	Lysinibacillus sphaericus
0.015	-	DL-propargylglycine	mutations Q18H/I336F/E313G	Lysinibacillus sphaericus
0.0176	-	DL-propargylglycine	mutations V135I/I308N/Q363H	Lysinibacillus sphaericus
0.0183	-	DL-propargylglycine	mutations F110Y/G124C/G293A	Lysinibacillus sphaericus
0.022	-	DL-propargylglycine	mutations D209G/Q18H/I336F	Lysinibacillus sphaericus
0.025	-	DL-propargylglycine	mutation G124A	Lysinibacillus sphaericus
0.032	-	DL-propargylglycine	mutations Q18H/I336F	Lysinibacillus sphaericus
0.032	-	DL-propargylglycine	mutations V33A/A206D/L283F	Lysinibacillus sphaericus
0.034	-	DL-propargylglycine	mutations K224Q/L283F	Lysinibacillus sphaericus
0.0384	-	DL-propargylglycine	mutation E313G	Lysinibacillus sphaericus
0.04	-	DL-propargylglycine	mutations G124A/E313G	Lysinibacillus sphaericus
0.33	-	L-phenylalanine	mutation G124A	Lysinibacillus sphaericus
0.49	-	L-phenylalanine	mutations G124A/E313G	Lysinibacillus sphaericus
0.68	-	L-phenylalanine	mutations G124A/E313N	Lysinibacillus sphaericus
0.74	-	L-phenylalanine	mutations F110Y/G124C/G293A	Lysinibacillus sphaericus
17.4	-	L-phenylalanine	mutations V135I/I308N/Q363H	Lysinibacillus sphaericus
40.5	-	L-phenylalanine	wild type enzyme of Bacillus sphaericus	Lysinibacillus sphaericus
50.6	-	L-phenylalanine	mutations Q18H/I336F/E313G	Lysinibacillus sphaericus
55.6	-	L-phenylalanine	mutations K224Q/L283F/E313G	Lysinibacillus sphaericus
58.3	-	L-phenylalanine	mutations D209G/Q18H/I336F	Lysinibacillus sphaericus
68.4	-	L-phenylalanine	mutations K224Q/L283F	Lysinibacillus sphaericus
84.9	-	L-phenylalanine	mutation D209G	Lysinibacillus sphaericus
91.6	-	L-phenylalanine	mutations Q18H/I336F	Lysinibacillus sphaericus
102.5	-	L-phenylalanine	mutations V33A/A206D/L283F	Lysinibacillus sphaericus
279	-	L-phenylalanine	mutation E313G	Lysinibacillus sphaericus