Expression and Characterization of Calcium- and Zinc-Tolerant Xylose Isomerase from Anoxybacillus kamchatkensis G10.
J Microbiol Biotechnol
; 28(4): 606-612, 2018 Apr 28.
Article
en En
| MEDLINE
| ID: mdl-29429321
ABSTRACT
The enzyme xylose isomerase (E.C. 5.3.1.5, XI) is responsible for the conversion of an aldose to ketose, especially xylose to xylulose. Owing to the ability of XI to isomerize glucose to fructose, this enzyme is used in the food industry to prepare high-fructose corn syrup. Therefore, we studied the characteristics of XI from Anoxybacillus kamchatkensis G10, a thermophilic bacterium. First, the gene coding for XI (xylA) was inserted into the pET-21a(+) expression vector and the construct was transformed into the Escherichia coli competent cell BL21 (DE3). The expression of recombinant XI was induced in the absence of isopropyl-thio-ß-galactopyranoside and purified using Ni-NTA affinity chromatography. The optimum temperature of recombinant XI was 80°C and measurement of the heat stability indicated that 55% of residual activity was maintained after 2 h incubation at 60°C. The optimum pH was found to be 7.5 in sodium phosphate buffer. Magnesium, manganese, and cobalt ions were found to increase the enzyme activity; manganese was the most effective. Additionally, recombinant XI was resistant to the presence of Ca²âº and Zn²âº ions. The kinetic properties, Km and Vmax, were calculated as 81.44 mM and 2.237 µmol/min/mg, respectively. Through redundancy analysis, XI of A. kamchatkensis G10 was classified into a family containing type II XIs produced by the genera Geobacillus, Bacillus, and Thermotoga. These results suggested that the thermostable nature of XI of A. kamchatkensis G10 may be advantageous in industrial applications and food processing.
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Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
Zinc
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Regulación Bacteriana de la Expresión Génica
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Calcio
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Isomerasas Aldosa-Cetosa
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Anoxybacillus
Idioma:
En
Revista:
J Microbiol Biotechnol
Año:
2018
Tipo del documento:
Article
Pais de publicación:
COREA DEL SUR
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CORÉIA DO SUL
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KR
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SOUTH KOREA