Literature summary for 1.4.1.B2 extracted from

Baker, J.J.; van der Drift, C.
Purification and properties of L-erythro-3,5-diaminohexanoate dehydrogenase from Clostridium sticklandii (1974), Biochemistry, 13, 292-299.

Inhibitors

Inhibitors	Comment	Organism
2,4-Diaminopentanoate	-	Acetoanaerobium sticklandii
2,5-Diaminohexanoate	-	Acetoanaerobium sticklandii
5-amino-3-oxohexanoate	competitive inhibition	Acetoanaerobium sticklandii
ADP	about half as effective as ATP	Acetoanaerobium sticklandii
AMP	32% as effective as ATP	Acetoanaerobium sticklandii
arsenate	addition of any of the neutral salts causes a parabolic inhibition. A direct comparison of arsenate and chloride ion shows that arsenate is not as inhibitory even though it has a higher ionic strength. Arsenate interacts with the enzyme differently from chloride and bromide. Sulfate, which is a large ion like arsenate, is as inhibitory as chloride (at equivalent ionic strength) with NAD+ as coenzyme, but it is much less effective than chloride with NADP+ as coenzyme	Acetoanaerobium sticklandii
ATP	competitive with respect to L-erythro-3,5-diaminohexanoate and NAD+	Acetoanaerobium sticklandii
Br-	addition of any of the neutral salts causes a parabolic inhibition. A direct comparison of arsenate and chloride ion shows that arsenate is not as inhibitory even though it has a higher ionic strength. Arsenate interacts with the enzyme differently from chloride and bromide. Sulfate, which is a large ion like arsenate, is as inhibitory as chloride (at equivalent ionic strength) with NAD+ as coenzyme, but it is much less effective than chloride with NADP+ as coenzyme	Acetoanaerobium sticklandii
Ca2+	0.5 mM, 22% inhibition at pH 7.8, cofactor NADP+, activation at pH 8.9	Acetoanaerobium sticklandii
Cl-	addition of any of the neutral salts causes a parabolic inhibition. A direct comparison of arsenate and chloride ion shows that arsenate is not as inhibitory even though it has a higher ionic strength. Arsenate interacts with the enzyme differently from chloride and bromide. Sulfate, which is a large ion like arsenate, is as inhibitory as chloride (at equivalent ionic strength) with NAD+ as coenzyme, but it is much less effective than chloride with NADP+ as coenzyme	Acetoanaerobium sticklandii
Co2+	0.5 mM, 71% inhibition at pH 7.8, cofactor NADP+, activation at pH 8.9	Acetoanaerobium sticklandii
CTP	about half as effective as ATP	Acetoanaerobium sticklandii
Cu2+	0.5 mM, 35% inhibition at pH 7.8, 30% inhibition at pH 8.9, cofactor NADP+	Acetoanaerobium sticklandii
D-erythro-3,5-diaminohexanoate	-	Acetoanaerobium sticklandii
D-Lysine	-	Acetoanaerobium sticklandii
DL-beta-aminobutyrate	-	Acetoanaerobium sticklandii
Fe2+	0.5 mM, 36% inhibition at pH 7.8, 9% inhibition at pH 8.9, cofactor NADP+	Acetoanaerobium sticklandii
Fe3+	0.5 mM, 8% inhibition at pH 8.9, no effect on activity at pH 7.8	Acetoanaerobium sticklandii
GTP	inhibits as effectively as ATP	Acetoanaerobium sticklandii
L-beta-Lysine	-	Acetoanaerobium sticklandii
L-lysine	-	Acetoanaerobium sticklandii
L-ornithine	-	Acetoanaerobium sticklandii
Mg2+	0.5 mM, 63% inhibition at pH 7.8, cofactor NADP+, activation at pH 8.9	Acetoanaerobium sticklandii
Mn2+	0.5 mM, 52% inhibition at pH 7.8, cofactor NADP+, activation at pH 8.9	Acetoanaerobium sticklandii
additional information	the lactams of DL-erythro-3,5-diaminohexanoate and DL-threo-3,5-diaminohexanoatae and 2-methylpyrrolidone-5-carboxylic acid have no effect on activity. Acetate (10 mM), butyrate (10 mM), acetyl phosphate (2 mM), or acetyl-CoA (0.62 mM) have no effect on enzyme activity	Acetoanaerobium sticklandii
NADH	competitive inhibition	Acetoanaerobium sticklandii
NADPH	competitive inhibition	Acetoanaerobium sticklandii
NH4+	10 mM NH4Cl and higher, inhibits the reaction, probably due to the effect of chloride on the activity	Acetoanaerobium sticklandii
SO42-	addition of any of the neutral salts causes a parabolic inhibition. A direct comparison of arsenate and chloride ion shows that arsenate is not as inhibitory even though it has a higher ionic strength. Arsenate interacts with the enzyme differently from chloride and bromide. Sulfate, which is a large ion like arsenate, is as inhibitory as chloride (at equivalent ionic strength) with NAD+ as coenzyme, but it is much less effective than chloride with NADP+ as coenzyme	Acetoanaerobium sticklandii
UTP	16% as effective as ATP	Acetoanaerobium sticklandii
Zn2+	0.5 mM, 49% inhibition at pH 7.8, cofactor NADP+, activation at pH 8.9	Acetoanaerobium sticklandii

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism
0.03	-	NADP+	arsenate buffer, pH 7.4, two apparent Km values (0.03 mM and 0.07 mM) are determined for NADP+ because the Lineweaver-Burk plot is biphasic	Acetoanaerobium sticklandii
0.07	-	NADP+	arsenate buffer, pH 7.4, two apparent Km values (0.03 mM and 0.07 mM) are determined for NADP+ because the Lineweaver-Burk plot is biphasic	Acetoanaerobium sticklandii
0.07	-	NADP+	arsenate buffer, pH 9.1, two apparent Km values (0.07 mM and 0.27 mM) are determined for NADP+ because the Lineweaver-Burk plot is biphasic	Acetoanaerobium sticklandii
0.08	-	L-erythro-3,5-diaminohexanoate	pH 7.4, in presence of 0.4 mM NADP+	Acetoanaerobium sticklandii
0.16	-	L-erythro-3,5-diaminohexanoate	pH 7.4, in presence of 0.4 mM NAD+	Acetoanaerobium sticklandii
0.27	-	NADP+	arsenate buffer, pH 9.1, two apparent Km values (0.07 mM and 0.27 mM) are determined for NADP+ because the Lineweaver-Burk plot is biphasic	Acetoanaerobium sticklandii
0.48	-	NAD+	arsenate buffer, pH 7.4	Acetoanaerobium sticklandii
0.67	-	NAD+	arsenate buffer, pH 9.1	Acetoanaerobium sticklandii
3.3	-	acetylpyridine-NAD+	pH 8.8	Acetoanaerobium sticklandii
22	-	deamino-NAD+	pH 8.8	Acetoanaerobium sticklandii

Metals/Ions

Metals/Ions	Comment	Organism
Ca2+	5 mM, 2.3fold activation at pH 8.9, inhibition at pH 7.8	Acetoanaerobium sticklandii
Co2+	5 mM, 1.5fold activation at pH 8.9, inhibition at pH 7.8	Acetoanaerobium sticklandii
Mn2+	at pH 8.9 Mn2+ gives the highest increase in activity (232% at 0.5 mM). Concentrations of 0.01, 0.05, 0.1, 0.75, and 1.0 mM increase the enzyme activity by 110, 156, 188, 258, and 263%, respectively. The activity is maximal at 0.75 mM Mn2+	Acetoanaerobium sticklandii
Zn2+	5 mM, 1.6fold activation at pH 8.9, inhibition at pH 7.8	Acetoanaerobium sticklandii

Molecular Weight [Da]

Molecular Weight [Da]	Molecular Weight Maximum [Da]	Comment	Organism
39300	-	2 * 39300, SDS-PAGE	Acetoanaerobium sticklandii
82000	-	gel filtration	Acetoanaerobium sticklandii

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
L-erythro-3,5-diaminohexanoate + H2O + NAD+	Acetoanaerobium sticklandii	either NAD+ or NADP+ serve as coenzyme. Double-reciprocal plots of activity versus pyridine nucleotide concentration are biphasic. The Km values for NADP+ are lower but a higher Vmax is observed with NAD+	5-amino-3-oxohexanoate + NH3 + NADH + H+	-	?
L-erythro-3,5-diaminohexanoate + H2O + NADP+	Acetoanaerobium sticklandii	either NAD+ or NADP+ serve as coenzyme. Double-reciprocal plots of activity versus pyridine nucleotide concentration are biphasic. The Km values for NADP+ are lower but a higher Vmax is observed with NAD+	5-amino-3-oxohexanoate + NH3 + NADPH + H+	-	?

Organism

Organism	UniProt	Comment	Textmining
Acetoanaerobium sticklandii	-	-	-

Purification (Commentary)

Purification (Comment)	Organism
-	Acetoanaerobium sticklandii

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg]	Specific Activity Maximum [µmol/min/mg]	Comment	Organism
15.9	-	-	Acetoanaerobium sticklandii

Storage Stability

Storage Stability	Organism
-20°C, 1 month, no loss of activity in crude enzyme extract	Acetoanaerobium sticklandii
23°C, 1 day, no loss of activity in crude enzyme extract	Acetoanaerobium sticklandii
4°C, 1 week, no loss of activity in crude enzyme extract	Acetoanaerobium sticklandii

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
L-erythro-3,5-diaminohexanoate + H2O + acetylpyridine-NAD+	-	Acetoanaerobium sticklandii	5-amino-3-oxohexanoate + NH3 + acetylpyridine-NADH + H+	-	?
L-erythro-3,5-diaminohexanoate + H2O + deamino-NAD+	-	Acetoanaerobium sticklandii	5-amino-3-oxohexanoate + NH3 + deamino-NADH + H+	-	?
L-erythro-3,5-diaminohexanoate + H2O + NAD+	either NAD+ or NADP+ serve as coenzyme. Double-reciprocal plots of activity versus pyridine nucleotide concentration are biphasic. The Km values for NADP+ are lower but a higher Vmax is observed with NAD+	Acetoanaerobium sticklandii	5-amino-3-oxohexanoate + NH3 + NADH + H+	-	?
L-erythro-3,5-diaminohexanoate + H2O + NAD+	Vmax/Km for NADP+ is 10-16fold higher, depending on KM-value	Acetoanaerobium sticklandii	5-amino-3-oxohexanoate + NH3 + NADH + H+	-	?
L-erythro-3,5-diaminohexanoate + H2O + NADP+	either NAD+ or NADP+ serve as coenzyme. Double-reciprocal plots of activity versus pyridine nucleotide concentration are biphasic. The Km values for NADP+ are lower but a higher Vmax is observed with NAD+	Acetoanaerobium sticklandii	5-amino-3-oxohexanoate + NH3 + NADPH + H+	-	?
L-erythro-3,5-diaminohexanoate + H2O + NADP+	Vmax/Km for NADP+ is 10-16fold higher, depending on KM-value	Acetoanaerobium sticklandii	5-amino-3-oxohexanoate + NH3 + NADPH + H+	-	?

Subunits

Subunits	Comment	Organism
dimer	2 * 39300, SDS-PAGE	Acetoanaerobium sticklandii

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
24	40	activity with NAD+, the activity of the enzyme increases up to 24°C, is relatively constant until 40°C and then decreases	Acetoanaerobium sticklandii
52	-	activity with NADP+ increases up to 52°C and then rapidly falls off, probably due to heat inactivation of the protein	Acetoanaerobium sticklandii

Temperature Range [°C]

Temperature Minimum [°C]	Temperature Maximum [°C]	Comment	Organism
25	59	25°C: about 40% of maximal activity, 50°C: about 70% of maximal activity, cofactor: NAD+	Acetoanaerobium sticklandii

Temperature Stability [°C]

Temperature Stability Minimum [°C]	Temperature Stability Maximum [°C]	Comment	Organism
40	-	30 min, no loss of activity	Acetoanaerobium sticklandii
50	-	t1/2: 56 min, inactivation follows first-order kinetics	Acetoanaerobium sticklandii
55	-	t1/2: 32 min, inactivation follows first-order kinetics. In the presence of both NAD+ and 3,5-diaminohexanoate, the enzyme is effectively stabilized against heat inactivation at 55°C. NAD+ alone accelerates heat inactivation	Acetoanaerobium sticklandii
59	-	t1/2: 4 min, inactivation follows first-order kinetics	Acetoanaerobium sticklandii

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
8.7	9	activity with NAD+ or NADP+	Acetoanaerobium sticklandii

pH Range

pH Minimum	pH Maximum	Comment	Organism
7.3	9.5	pH 7.3: about 45% of maximal activity, pH 9.5: about 80% of maximal activity, cofactor: NAD+	Acetoanaerobium sticklandii
7.8	10.2	pH 7.8: about 55% of maximal activity, pH 10.2: about 55% of maximal activity, cofactor: NADP+	Acetoanaerobium sticklandii

pH Stability

pH Stability	pH Stability Maximum	Comment	Organism
5	10.5	25°C, 30 min, no loss of activity	Acetoanaerobium sticklandii

Cofactor

Cofactor	Comment	Organism
acetylpyridine-NAD+	-	Acetoanaerobium sticklandii
deamino-NAD+	-	Acetoanaerobium sticklandii
NAD+	Vmax/Km for NADP+ is 10-16fold higher, depending on KM-value	Acetoanaerobium sticklandii
NADP+	Vmax/Km for NADP+ is 10-16fold higher, depending on KM-value	Acetoanaerobium sticklandii

Ki Value [mM]

Ki Value [mM]	Ki Value maximum [mM]	Inhibitor	Comment	Organism
0.0005	-	NADPH	pH 7.4	Acetoanaerobium sticklandii
0.0195	-	NADH	pH 7.4	Acetoanaerobium sticklandii
0.1	-	D-erythro-3,5-diaminohexanoate	pH 7.4	Acetoanaerobium sticklandii
0.19	-	2,5-Diaminohexanoate	pH 7.4	Acetoanaerobium sticklandii
0.52	-	2,4-Diaminopentanoate	pH 7.4	Acetoanaerobium sticklandii
1	-	ATP	pH 7.4, with DL-erythro-3,5-diaminohexanoate as variable substrate	Acetoanaerobium sticklandii
2.5	-	L-lysine	pH 7.4	Acetoanaerobium sticklandii
2.5	-	L-beta-Lysine	pH 7.4	Acetoanaerobium sticklandii
4.5	-	ATP	pH 7.4, with NAD+ as variable substrate	Acetoanaerobium sticklandii
5.4	-	L-ornithine	pH 7.4	Acetoanaerobium sticklandii
11.3	-	D-Lysine	pH 7.4	Acetoanaerobium sticklandii
73.4	-	DL-beta-aminobutyrate	pH 7.4	Acetoanaerobium sticklandii