Literature summary for 5.3.1.8 extracted from

Yeom, S.J.; Seo, E.S.; Kim, B.N.; Kim, Y.S.; Oh, D.K.
Characterization of a mannose-6-phosphate isomerase from Thermus thermophilus and increased L-ribose production by its R142N mutant (2011), Appl. Environ. Microbiol., 77, 762-767.

Cloned(Commentary)

Cloned (Comment)	Organism
expressed in Escherichia coli ER2566 cells	Thermus thermophilus

Protein Variants

Protein Variants	Comment	Organism
D138A	the enzyme shows decreased specific activity for L-ribulose	Thermus thermophilus
E132A	the mutants has no activity for L-ribulose	Thermus thermophilus
E67A	the mutants has no activity for L-ribulose	Thermus thermophilus
H122A	the mutants has no activity for L-ribulose	Thermus thermophilus
H50A	the mutants has no activity for L-ribulose	Thermus thermophilus
K37A	the enzyme shows decreased specific activity for L-ribulose	Thermus thermophilus
K65A	the enzyme shows decreased specific activity for L-ribulose	Thermus thermophilus
L124A	the enzyme shows decreased specific activity for L-ribulose	Thermus thermophilus
L18A	the enzyme shows decreased specific activity for L-ribulose	Thermus thermophilus
L39A	the enzyme shows decreased specific activity for L-ribulose	Thermus thermophilus
Q48A	the enzyme shows decreased specific activity for L-ribulose	Thermus thermophilus
R11A	the enzyme shows about wild type specific activity for L-ribulose	Thermus thermophilus
R142A	the enzyme shows increased specific activity for L-ribulose	Thermus thermophilus
R142E	the enzyme shows decreased specific activity for L-ribulose	Thermus thermophilus
R142K	the enzyme shows decreased specific activity for L-ribulose	Thermus thermophilus
R142N	the specific activity and catalytic efficiency (kcat/Km) for L-ribulose using the R142N mutant are 1.4 and 1.6fold higher than those of the wild type enzyme, respectively	Thermus thermophilus
R142Q	the enzyme shows decreased specific activity for L-ribulose	Thermus thermophilus
R142Y	the enzyme shows decreased specific activity for L-ribulose	Thermus thermophilus
W13A	the enzyme shows decreased specific activity for L-ribulose	Thermus thermophilus
W69A	the enzyme shows decreased specific activity for L-ribulose	Thermus thermophilus

Inhibitors

Inhibitors	Comment	Organism	Structure
EDTA	about 75% residual activity at 0.5 mM	Thermus thermophilus
Fe2+	12% residual activity at 0.5 mM	Thermus thermophilus

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism
136	-	L-ribulose	wild type enzyme, at pH 7.0 and 75°C	Thermus thermophilus
140	-	L-ribulose	mutant enzyme R142N, at pH 7.0 and 75°C	Thermus thermophilus
150	-	L-ribulose	mutant enzyme R142Q, at pH 7.0 and 75°C	Thermus thermophilus
151	-	L-ribulose	mutant enzyme R142E, at pH 7.0 and 75°C	Thermus thermophilus
184	-	L-ribulose	mutant enzyme R142A, at pH 7.0 and 75°C	Thermus thermophilus
228	-	L-ribulose	mutant enzyme R142K, at pH 7.0 and 75°C	Thermus thermophilus
308	-	L-ribulose	mutant enzyme R142Y, at pH 7.0 and 75°C	Thermus thermophilus

Metals/Ions

Metals/Ions	Comment	Organism
Co2+	about 230% activity at 0.5 mM	Thermus thermophilus
Cu2+	maximal activity of the recombinant enzyme for L-ribulose isomerization in the presence of 0.5 mM Cu2+	Thermus thermophilus
Mn2+	about 160% activity at 0.5 mM	Thermus thermophilus
additional information	the enzyme is not affected by Ca2+, Mg2+, Ba2+, and Ni2+	Thermus thermophilus
Zn2+	about 130% activity at 0.5 mM	Thermus thermophilus

Molecular Weight [Da]

Molecular Weight [Da]	Molecular Weight Maximum [Da]	Comment	Organism
29000	-	gel filtration	Thermus thermophilus
29054	-	1 * 29054, calculated from amino acid sequence	Thermus thermophilus

Organism

Organism	UniProt	Comment	Textmining
Thermus thermophilus	-	-	-
Thermus thermophilus KCCM 40879	-	-	-

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg]	Specific Activity Maximum [µmol/min/mg]	Comment	Organism
3	-	wild type enzyme, using D-psicose as substrate, at pH 7.0 and 75°C	Thermus thermophilus
16	-	wild type enzyme, using L-tagatose as substrate, at pH 7.0 and 75°C	Thermus thermophilus
19	-	wild type enzyme, using L-mannose as substrate, at pH 7.0 and 75°C	Thermus thermophilus
22	-	wild type enzyme, using L-xylulose as substrate, at pH 7.0 and 75°C	Thermus thermophilus
23	-	wild type enzyme, using L-lyxose as substrate, at pH 7.0 and 75°C	Thermus thermophilus
42	-	wild type enzyme, using L-psicose as substrate, at pH 7.0 and 75°C	Thermus thermophilus
57	-	wild type enzyme, using L-fructose as substrate, at pH 7.0 and 75°C	Thermus thermophilus
58	-	wild type enzyme, using D-ribose as substrate, at pH 7.0 and 75°C	Thermus thermophilus
63	-	wild type enzyme, using L-talose as substrate, at pH 7.0 and 75°C	Thermus thermophilus
85	-	wild type enzyme, using D-tagatose as substrate, at pH 7.0 and 75°C	Thermus thermophilus
126	-	wild type enzyme, using D-ribulose as substrate, at pH 7.0 and 75°C	Thermus thermophilus
128	-	wild type enzyme, using D-xylulose as substrate, at pH 7.0 and 75°C	Thermus thermophilus
165	-	wild type enzyme, using D-allose as substrate, at pH 7.0 and 75°C	Thermus thermophilus
167	-	wild type enzyme, using D-fructose as substrate, at pH 7.0 and 75°C	Thermus thermophilus
333	-	wild type enzyme, using D-lyxose as substrate, at pH 7.0 and 75°C	Thermus thermophilus
425	-	wild type enzyme, using L-ribose as substrate, at pH 7.0 and 75°C	Thermus thermophilus
474	-	mutant enzyme K37A, using L-ribulose as substrate, at pH 7.0 and 75°C	Thermus thermophilus
742	-	wild type enzyme, using L-allose as substrate, at pH 7.0 and 75°C	Thermus thermophilus
838	-	mutant enzyme W13A, using L-ribulose as substrate, at pH 7.0 and 75°C	Thermus thermophilus
842	-	wild type enzyme, using D-mannose as substrate, at pH 7.0 and 75°C	Thermus thermophilus
972	-	mutant enzyme D138A, using L-ribulose as substrate, at pH 7.0 and 75°C	Thermus thermophilus
1011	-	mutant enzyme K65A, using L-ribulose as substrate, at pH 7.0 and 75°C	Thermus thermophilus
1016	-	mutant enzyme Q48A, using L-ribulose as substrate, at pH 7.0 and 75°C	Thermus thermophilus
1045	-	mutant enzyme R142E, using L-ribulose as substrate, at pH 7.0 and 75°C	Thermus thermophilus
1046	-	mutant enzyme W69A, using L-ribulose as substrate, at pH 7.0 and 75°C	Thermus thermophilus
1065	-	mutant enzyme L39A, using L-ribulose as substrate, at pH 7.0 and 75°C	Thermus thermophilus
1076	-	mutant enzyme L124A, using L-ribulose as substrate, at pH 7.0 and 75°C	Thermus thermophilus
1092	-	mutant enzyme R142Y, using L-ribulose as substrate, at pH 7.0 and 75°C	Thermus thermophilus
1194	-	mutant enzyme L18A, using L-ribulose as substrate, at pH 7.0 and 75°C	Thermus thermophilus
1214	-	mutant enzyme R142K, using L-ribulose as substrate, at pH 7.0 and 75°C	Thermus thermophilus
1270	-	mutant enzyme R142Q, using L-ribulose as substrate, at pH 7.0 and 75°C	Thermus thermophilus
1482	-	mutant enzyme R11A, using L-ribulose as substrate, at pH 7.0 and 75°C	Thermus thermophilus
1493	-	wild type enzyme, using L-ribulose as substrate, at pH 7.0 and 75°C	Thermus thermophilus
1619	-	wild type enzyme, using D-talose as substrate, at pH 7.0 and 75°C	Thermus thermophilus
1791	-	mutant enzyme R142A, using L-ribulose as substrate, at pH 7.0 and 75°C	Thermus thermophilus
2152	-	mutant enzyme R14N, using L-ribulose as substrate, at pH 7.0 and 75°C	Thermus thermophilus

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
D-allose	-	Thermus thermophilus	D-psicose	-	r
D-allose	-	Thermus thermophilus KCCM 40879	D-psicose	-	r
D-Lyxose	-	Thermus thermophilus	D-Xylulose	-	r
D-Lyxose	-	Thermus thermophilus KCCM 40879	D-Xylulose	-	r
D-Mannose	-	Thermus thermophilus	D-Fructose	-	r
D-ribose	-	Thermus thermophilus	D-ribulose	-	r
D-ribose	-	Thermus thermophilus KCCM 40879	D-ribulose	-	r
D-talose	-	Thermus thermophilus	D-tagatose	-	r
L-allose	-	Thermus thermophilus	L-psicose	-	r
L-lyxose	-	Thermus thermophilus	L-xylulose	-	r
L-lyxose	-	Thermus thermophilus KCCM 40879	L-xylulose	-	r
L-Mannose	-	Thermus thermophilus	L-Fructose	-	r
L-Mannose	-	Thermus thermophilus KCCM 40879	L-Fructose	-	r
L-ribose	-	Thermus thermophilus	L-ribulose	-	r
L-ribulose	highest activity	Thermus thermophilus	L-ribose	-	r
L-talose	-	Thermus thermophilus	L-tagatose	-	r

Subunits

Subunits	Comment	Organism
monomer	1 * 29000, SDS-PAGE	Thermus thermophilus
monomer	1 * 29054, calculated from amino acid sequence	Thermus thermophilus

Synonyms

Synonyms	Comment	Organism
MPI	-	Thermus thermophilus

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
75	-	maximal activity of the recombinant enzyme for L-ribulose isomerization	Thermus thermophilus

Temperature Stability [°C]

Temperature Stability Minimum [°C]	Temperature Stability Maximum [°C]	Comment	Organism
65	85	the enzyme shows half-lives of 22, 10, 5.5, 2.1, and 0.3 h at 65, 70, 75, 80, and 85°C, respectively	Thermus thermophilus

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism
32670	-	L-ribulose	mutant enzyme R142E, at pH 7.0 and 75°C	Thermus thermophilus
48300	-	L-ribulose	mutant enzyme R142Q, at pH 7.0 and 75°C	Thermus thermophilus
50640	-	L-ribulose	wild type enzyme, at pH 7.0 and 75°C	Thermus thermophilus
56180	-	L-ribulose	mutant enzyme R142Y, at pH 7.0 and 75°C	Thermus thermophilus
64870	-	L-ribulose	mutant enzyme R142A, at pH 7.0 and 75°C	Thermus thermophilus
68880	-	L-ribulose	mutant enzyme R142K, at pH 7.0 and 75°C	Thermus thermophilus
81060	-	L-ribulose	mutant enzyme R142N, at pH 7.0 and 75°C	Thermus thermophilus

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
7	-	maximal activity of the recombinant enzyme for L-ribulose isomerization	Thermus thermophilus

kcat/KM [mM/s]

kcat/KM Value [1/mMs^-1]	kcat/KM Value Maximum [1/mMs^-1]	Substrate	Comment	Organism
182	-	L-ribulose	mutant enzyme R142Y, at pH 7.0 and 75°C	Thermus thermophilus
216	-	L-ribulose	mutant enzyme R142E, at pH 7.0 and 75°C	Thermus thermophilus
302	-	L-ribulose	mutant enzyme R142K, at pH 7.0 and 75°C	Thermus thermophilus
322	-	L-ribulose	mutant enzyme R142Q, at pH 7.0 and 75°C	Thermus thermophilus
353	-	L-ribulose	mutant enzyme R142A, at pH 7.0 and 75°C	Thermus thermophilus
374	-	L-ribulose	wild type enzyme, at pH 7.0 and 75°C	Thermus thermophilus
579	-	L-ribulose	mutant enzyme R142N, at pH 7.0 and 75°C	Thermus thermophilus