Literature summary for 5.3.1.4 extracted from

Rhimi, M.; Ilhammami, R.; Bajic, G.; Boudebbouze, S.; Maguin, E.; Haser, R.; Aghajari, N.
The acid tolerant L-arabinose isomerase from the food grade Lactobacillus sakei 23K is an attractive D-tagatose producer (2010), Biores. Technol., 101, 9171-9177.

Search for more literature for 5.3.1.4

Application

Application	Comment	Organism
synthesis	the enzyme is used for the commercial production of D-tagatose	Latilactobacillus sakei

Cloned(Commentary)

Cloned (Comment)	Organism
expressed in Escherichia coli BL21/pMR36 (called MRS36) cells	Latilactobacillus sakei

Inhibitors

Inhibitors	Comment	Organism	Structure
Cu2+	about 30% residual activity at 1 mM	Latilactobacillus sakei
EDTA	54% inhibition at 1 mM EDTA assayed at 35°C	Latilactobacillus sakei

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
31.6	-	L-arabinose	with 0.8 mM MgCl2 and 0.8 mM MnCl2, in 100 mM sodium acetate buffer (pH 5.0), at 35°C	Latilactobacillus sakei
59	-	D-galactose	with 0.8 mM MgCl2 and 0.8 mM MnCl2, in 100 mM sodium acetate buffer (pH 5.0), at 35°C	Latilactobacillus sakei

Metals/Ions

Metals/Ions	Comment	Organism	Structure
Mg2+	the enzyme has low metal requirement of only 0.8 mM Mg2+ for its maximal activity and thermostability. At 35°C, the addition of 1 mM Mg2+ to the EDTA-treated enzyme increases the specific activity from 65 to 201 units/mg	Latilactobacillus sakei
Mn2+	the enzyme has low metal requirement of only 0.8 mM Mn2+ for its maximal activity and thermostability (above 35°C). At 35°C, the addition of 1 mM Mn2+ to the EDTA-treated enzyme increases the specific activity from 65 to 204 units/mg	Latilactobacillus sakei
additional information	the enzyme is not affected by Zn2+, Ca2+, Co2+, Fe2+, and Ba2+	Latilactobacillus sakei

Molecular Weight [Da]

Molecular Weight [Da]	Molecular Weight Maximum [Da]	Comment	Organism
53680	-	4 * 53680, calculated from amino acid sequence	Latilactobacillus sakei
54000	-	4 * 54000, SDS-PAGE	Latilactobacillus sakei
220000	-	gel filtration	Latilactobacillus sakei

Organism

Organism	UniProt	Comment	Textmining
Latilactobacillus sakei	-	-	-
Latilactobacillus sakei 23K	-	-	-

Purification (Commentary)

Purification (Comment)	Organism
ammonium sulfate precipitation, Mono Q column chromatography, and Sephacryl S-200 gel filtration	Latilactobacillus sakei

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
D-galactose	36% conversion after 7 h at 40°C	Latilactobacillus sakei	D-tagatose	-	?
D-galactose	36% conversion after 7 h at 40°C	Latilactobacillus sakei 23K	D-tagatose	-	?
L-arabinose	-	Latilactobacillus sakei	L-ribulose	-	?
L-arabinose	-	Latilactobacillus sakei 23K	L-ribulose	-	?

Subunits

Subunits	Comment	Organism
homotetramer	4 * 54000, SDS-PAGE	Latilactobacillus sakei
homotetramer	4 * 53680, calculated from amino acid sequence	Latilactobacillus sakei

Synonyms

Synonyms	Comment	Organism
L-AI	-	Latilactobacillus sakei

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
30	40	-	Latilactobacillus sakei

Temperature Range [°C]

Temperature Minimum [°C]	Temperature Maximum [°C]	Comment	Organism
4	45	the enzyme is maximally active between 30 and 40°C is also active at low temperatures including 4, 10, 15, 20 and 25°C where the protein has nearly 70%, 72%, 75%, 85% and 90% of its maximal activity, respectively. At 45°C the enzyme retains 80% of its relative activity, but at higher temperatures the relative activity decreased rapidly	Latilactobacillus sakei

Temperature Stability [°C]

Temperature Stability Minimum [°C]	Temperature Stability Maximum [°C]	Comment	Organism
35	50	when treated with EDTA, the enzym is fully stable at temperatures up to 35°C for 24 h, but after 2 h at 40 and 45°C the enzyme retains only 95% and 55% of its activity, respectively, and at 50°C the half-life is evaluated to 80 min. At higher temperatures the enzyme is rapidly inactivated	Latilactobacillus sakei

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
5	7	-	Latilactobacillus sakei

pH Range

pH Minimum	pH Maximum	Comment	Organism
3	8	the enzyme preserves 85%, 89% and 95% at pH 3.5, 4 and 4.5, respectively, and more than 80% of its activity in the pH range from pH 3.0 to 8.0	Latilactobacillus sakei

pH Stability

pH Stability	pH Stability Maximum	Comment	Organism
3	6	at pH 4.5-6.0, the protein is highly stable. The enzyme displays significant half-lives of 33 and 36 h at pH 3.0 and 3.5, respectively	Latilactobacillus sakei

kcat/KM [mM/s]

kcat/KM Value [1/mMs^-1]	kcat/KM Value Maximum [1/mMs^-1]	Substrate	Comment	Organism	Structure
0.17	-	D-galactose	with 0.8 mM MgCl2 and 0.8 mM MnCl2, in 100 mM sodium acetate buffer (pH 5.0), at 35°C	Latilactobacillus sakei
1.08	-	L-arabinose	with 0.8 mM MgCl2 and 0.8 mM MnCl2, in 100 mM sodium acetate buffer (pH 5.0), at 35°C	Latilactobacillus sakei

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Release 2023.1 (January 2023)