Literature summary for 5.3.1.5 extracted from

Harris, J.M.; Epting, K.L.; Kelly, R.M.
N-terminal fusion of a hyperthermophilic chitin-binding domain to xylose isomerase from Thermotoga neapolitana enhances kinetics and thermostability of both free and immobilized enzymes (2010), Biotechnol. Prog., 26, 993-1000.

Search for more literature for 5.3.1.5

Cloned(Commentary)

Cloned (Comment)	Organism
expressed in Escherichia coli BL21(DE3) cells	Thermotoga neapolitana

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
65.2	-	D-glucose	soluble wild type enzyme, at 80°C, pH 7.0	Thermotoga neapolitana
88.5	-	D-glucose	soluble wild type enzyme, at 90°C, pH 7.0	Thermotoga neapolitana
121.5	-	D-glucose	immobilized CBD-TNX fusion protein, at 80°C, pH 7.0	Thermotoga neapolitana
148.9	-	D-glucose	immobilized CBD-TNX fusion protein, at 90°C, pH 7.0	Thermotoga neapolitana
157.8	-	D-glucose	unbound CBD-TNX fusion protein, at 80°C, pH 7.0	Thermotoga neapolitana
536.1	-	D-glucose	unbound CBD-TNX fusion protein, at 90°C, pH 7.0	Thermotoga neapolitana

Metals/Ions

Metals/Ions	Comment	Organism	Structure
Co2+	required for thermostability	Thermotoga neapolitana
Mn2+	required for thermostability	Thermotoga neapolitana

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
D-Xylose	Thermotoga neapolitana	-	D-Xylulose	-	?
D-Xylose	Thermotoga neapolitana 5068	-	D-Xylulose	-	?

Organism

Organism	UniProt	Comment	Textmining
Thermotoga neapolitana	-	-	-
Thermotoga neapolitana 5068	-	-	-

Purification (Commentary)

Purification (Comment)	Organism
heat treatment, DEAE-Sepharose column chromatography and Q-Sepharose column chromatography	Thermotoga neapolitana

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
D-Glucose	-	Thermotoga neapolitana	D-Fructose	-	?
D-Glucose	-	Thermotoga neapolitana 5068	D-Fructose	-	?
D-Xylose	-	Thermotoga neapolitana	D-Xylulose	-	?
D-Xylose	-	Thermotoga neapolitana 5068	D-Xylulose	-	?

Synonyms

Synonyms	Comment	Organism
XYLA	-	Thermotoga neapolitana

Temperature Stability [°C]

Temperature Stability Minimum [°C]	Temperature Stability Maximum [°C]	Comment	Organism
97	112	chitin-binding domain-D-xylose isomerase fusion protein (CBD-TNXI) bound to chitin has a half-life approximately 3times longer than the soluble wild type xylose isomerase (19.9 h vs. 6.8 h, respectively). The unbound soluble CBD-TNXI has a significantly longer half-life (56.5 h) than the immobilized enzyme. TNXI-apo enzyme melts at 97.5°C, while transitions at 100°C and 112°C are observed in the presence of Co2+ (0.5 mM) and Mn2+ (5.0 mM). The apo version of the immobilized enzyme melts at 104°C, with transitions at 87°C and 110°C for the halo enzyme	Thermotoga neapolitana

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism	Structure
16.2	-	D-glucose	soluble wild type enzyme, at 80°C, pH 7.0	Thermotoga neapolitana
19	-	D-glucose	soluble wild type enzyme, at 90°C, pH 7.0	Thermotoga neapolitana
63.3	-	D-glucose	unbound CBD-TNX fusion protein, at 80°C, pH 7.0	Thermotoga neapolitana
96.3	-	D-glucose	immobilized CBD-TNX fusion protein, at 80°C, pH 7.0	Thermotoga neapolitana
116	-	D-glucose	immobilized CBD-TNX fusion protein, at 90°C, pH 7.0	Thermotoga neapolitana
172.6	-	D-glucose	unbound CBD-TNX fusion protein, at 90°C, pH 7.0	Thermotoga neapolitana

kcat/KM [mM/s]

kcat/KM Value [1/mMs^-1]	kcat/KM Value Maximum [1/mMs^-1]	Substrate	Comment	Organism	Structure
0.22	-	D-glucose	soluble wild type enzyme, at 90°C, pH 7.0	Thermotoga neapolitana
0.25	-	D-glucose	soluble wild type enzyme, at 80°C, pH 7.0	Thermotoga neapolitana
0.32	-	D-glucose	unbound CBD-TNX fusion protein, at 90°C, pH 7.0	Thermotoga neapolitana
0.4	-	D-glucose	unbound CBD-TNX fusion protein, at 80°C, pH 7.0	Thermotoga neapolitana
0.78	-	D-glucose	immobilized CBD-TNX fusion protein, at 90°C, pH 7.0	Thermotoga neapolitana
0.79	-	D-glucose	immobilized CBD-TNX fusion protein, at 80°C, pH 7.0	Thermotoga neapolitana

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Release 2023.1 (January 2023)