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논문 기본 정보
- 자료유형
- 학술저널
- 저자정보
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- 한국미생물생명공학회 Journal of Microbiology and Biotechnology Journal of Microbiology and Biotechnology 제28권 제4호
- 발행연도
- 2018.1
- 수록면
- 606 - 612 (7page)
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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-β- galactopyranoside and purified using Ni-NTA affinity chromatography. The optimum temperature of recombinant XI was 80oC and measurement of the heat stability indicated that 55% of residual activity was maintained after 2 h incubation at 60oC. 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 Ca2+ and Zn2+ 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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참고문헌 (34)
참고문헌 신청
Bhosale SH / 1996 / Molecular and industrial aspects of glucose isomerase / Microbiol. Rev. 60:280~300
Tukel SS / 2008 / Catalytic efficiency of immobilized glucose isomerase in isomerization of glucose to fructose / Food Chem. 111:658~662
Ho N / 1983 / Purification and characterization of the $\small{D}$ -xylose isomerase gene from Escherichia coli / Enzyme Microb. Technol. 5:417~420
Dekker K / 1991 / Xylose (glucose) isomerase gene from the thermophile Thermus thermophilus: cloning, sequencing, and comparison with other thermostable xylose isomerases / J. Bacteriol. 173:3078~3083
Wilhelm M / 1984 / Selective cloning of Bacillus subtilis xylose isomerase and xylulokinase in Escherichia coli genes by IS5-mediated expression / EMBO J. 3:2555~2560
Tukel SS / 2008 / Catalytic efficiency of immobilized glucose isomerase in isomerization of glucose to fructose / Food Chem. 111:658~662
Ho N / 1983 / Purification and characterization of the
Dekker K / 1991 / Xylose (glucose) isomerase gene from the thermophile Thermus thermophilus: cloning, sequencing, and comparison with other thermostable xylose isomerases / J. Bacteriol. 173:3078~3083
Wilhelm M / 1984 / Selective cloning of Bacillus subtilis xylose isomerase and xylulokinase in Escherichia coli genes by IS5-mediated expression / EMBO J. 3:2555~2560
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