Literature summary for 2.6.1.9 extracted from

Marienhagen, J.; Sandalova, T.; Sahm, H.; Eggeling, L.; Schneider, G.
Insights into the structural basis of substrate recognition by histidinol-phosphate aminotransferase from Corynebacterium glutamicum (2008), Acta Crystallogr. Sect. D, 64, 675-685.

Crystallization (Commentary)

Crystallization (Comment)	Organism
apo histidinol-phosphate aminotransferase the internal PLP aldimine adduct and a pyridoxamine 5'-phosphate enzyme complex at resolutions of 2.2 A, 2.1 A and 1.8 A, respectively. The hydrogen bond between the side chain of residue Y21 and the phosphate group of histidinol phosphate is important for recognition of the natural substrate and discrimination against other potential amino donors. Residue N99 does not contribute to the specific recognition of the amino-acid donor. Residues Y123 and Y257 interact with the substrate through van der Waals-interactions	Corynebacterium glutamicum

Crystallization (Comment)

Organism

apo histidinol-phosphate aminotransferase the internal PLP aldimine adduct and a pyridoxamine 5'-phosphate enzyme complex at resolutions of 2.2 A, 2.1 A and 1.8 A, respectively. The hydrogen bond between the side chain of residue Y21 and the phosphate group of histidinol phosphate is important for recognition of the natural substrate and discrimination against other potential amino donors. Residue N99 does not contribute to the specific recognition of the amino-acid donor. Residues Y123 and Y257 interact with the substrate through van der Waals-interactions

Corynebacterium glutamicum

Protein Variants

Protein Variants	Comment	Organism
N99D	6fold reduction in ratio kcat to Km for substrates L-histidinol phosphate, 2-oxoglutarate	Corynebacterium glutamicum
Y123F	4fold reduction in ratio kcat to Km for substrates L-histidinol phosphate, 2-oxoglutarate	Corynebacterium glutamicum
Y21F	10fold reduction in ratio kcat to Km for substrates L-histidinol phosphate, 2-oxoglutarate	Corynebacterium glutamicum
Y257F	5fold reduction in ratio kcat to Km for substrates L-histidinol phosphate, 2-oxoglutarate	Corynebacterium glutamicum

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism
0.89	-	L-histidinol phosphate	wild-type	Corynebacterium glutamicum
1.3	-	L-histidinol phosphate	mutant Y123F	Corynebacterium glutamicum
2.2	-	L-histidinol phosphate	mutant Y257F	Corynebacterium glutamicum
6.7	-	L-histidinol phosphate	mutant Y21F	Corynebacterium glutamicum
9.1	-	L-histidinol phosphate	mutant N99D	Corynebacterium glutamicum
22.2	-	L-phenylalanine	mutant Y257F	Corynebacterium glutamicum
25.5	-	L-phenylalanine	mutant Y21F	Corynebacterium glutamicum
27.5	-	L-phenylalanine	mutant Y123F	Corynebacterium glutamicum
32.8	-	L-leucine	mutant Y257F	Corynebacterium glutamicum
53.4	-	L-leucine	mutant Y123F	Corynebacterium glutamicum
55.8	-	L-leucine	mutant N99D	Corynebacterium glutamicum
71.4	-	L-phenylalanine	mutant N99D	Corynebacterium glutamicum
80.8	-	L-leucine	mutant Y21F	Corynebacterium glutamicum
94.9	-	L-leucine	wild-type	Corynebacterium glutamicum
106	-	L-phenylalanine	wild-type	Corynebacterium glutamicum

Organism

Organism	UniProt	Comment	Textmining
Corynebacterium glutamicum	Q9KJU4	-	-

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg]	Specific Activity Maximum [µmol/min/mg]	Comment	Organism
0.1	-	substrates 4-methyl-2-oxopentanoate, L-alanine	Corynebacterium glutamicum
0.2	-	substrates L-glutarate, 2-oxobutanoate	Corynebacterium glutamicum
0.3	-	substrates 3-indole-2-oxopropanoate, L-glutamate	Corynebacterium glutamicum
0.4	-	substrates 4-methyl-2-oxopentanoate, L-asparagine	Corynebacterium glutamicum
0.5	-	substrates 3-(4-hydroxyphenyl)-2-oxopropanoate, L-glutamate	Corynebacterium glutamicum
1.4	-	substrates 4-methyl-2-oxopentanoate, L-glutamate	Corynebacterium glutamicum
1.5	-	substrates phenylpyruvate, L-glutamate	Corynebacterium glutamicum
2.8	-	substrates L-histidinol phosphate, 2-oxoglutarate	Corynebacterium glutamicum

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
3-(4-hydroxyphenyl)-2-oxopropanoate + L-glutamate	-	Corynebacterium glutamicum	L-tyrosine + 2-oxoglutarate	-	?
3-indole-2-oxopropanoate + L-glutamate	-	Corynebacterium glutamicum	L-tryptophan + 2-oxoglutarate	-	?
4-methyl-2-oxopentanoate + L-alanine	-	Corynebacterium glutamicum	L-leucine + pyruvate	-	?
4-methyl-2-oxopentanoate + L-asparagine	-	Corynebacterium glutamicum	L-leucine + 2-oxosuccinamic acid	-	?
4-methyl-2-oxopentanoate + L-glutamate	-	Corynebacterium glutamicum	L-leucine + 2-oxoglutarate	-	?
L-glutarate + 2-oxobutanoate	-	Corynebacterium glutamicum	2-oxoglutarate + 2-aminobutanoate	-	?
L-histidinol phosphate + 2-oxoglutarate	-	Corynebacterium glutamicum	3-(imidazol-4-yl)-2-oxopropyl phosphate + L-glutamate	-	?
additional information	no substrate: 3-methyl-2-oxopentanoate, 3-methyl-2-oxobutanoate	Corynebacterium glutamicum	?	-	?
phenylpyruvate + L-glutamate	-	Corynebacterium glutamicum	L-phenylalanine + 2-oxoglutarate	-	?

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism
0.39	-	L-histidinol phosphate	mutant Y123F	Corynebacterium glutamicum
0.51	-	L-histidinol phosphate	mutant Y257F	Corynebacterium glutamicum
0.59	-	L-phenylalanine	mutant Y123F	Corynebacterium glutamicum
0.66	-	L-leucine	mutant Y123F	Corynebacterium glutamicum
0.68	-	L-phenylalanine	mutant Y257F	Corynebacterium glutamicum
0.69	-	L-leucine	mutant Y257F	Corynebacterium glutamicum
0.87	-	L-phenylalanine	mutant Y21F	Corynebacterium glutamicum
1.18	-	L-histidinol phosphate	wild-type	Corynebacterium glutamicum
1.6	-	L-leucine	mutant N99D	Corynebacterium glutamicum
1.72	-	L-leucine	mutant Y21F	Corynebacterium glutamicum
1.86	-	L-histidinol phosphate	mutant N99D	Corynebacterium glutamicum
1.93	-	L-histidinol phosphate	mutant Y21F	Corynebacterium glutamicum
2.22	-	L-phenylalanine	mutant N99D	Corynebacterium glutamicum
3.27	-	L-leucine	wild-type	Corynebacterium glutamicum
4.1	-	L-phenylalanine	wild-type	Corynebacterium glutamicum