본문 바로가기
KERD
논문KCI 등재

Expression and Characterization of Calcium- and Zinc-Tolerant Xylose Isomerase from Anoxybacillus kamchatkensis G10

2

조회

0

다운로드

0

인용

0

참고문헌

키워드

주제분류생물학

초록

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.

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-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.

참고문헌 (34)

  1. Bhosale SH, "Molecular and industrial aspects of glucose isomerase", Microbiol. Rev., 60, pp.280~300 (1996)
  2. Tukel SS, "Catalytic efficiency of immobilized glucose isomerase in isomerization of glucose to fructose", Food Chem., 111, pp.658~662 (2008)[DOI]
  3. Ho N, "Purification and characterization of the <TEX>$\small{D}$<", TEX>-xylose isomerase gene from Escherichia coli, 1983, pp.5, 417~420
  4. Dekker K, "Xylose (glucose) isomerase gene from the thermophile Thermus thermophilus: cloning, sequencing, and comparison with other thermostable xylose isomerases", J. Bacteriol., 173, pp.3078~3083 (1991)[DOI]
  5. Wilhelm M, "Selective cloning of Bacillus subtilis xylose isomerase and xylulokinase in Escherichia coli genes by IS5-mediated expression", EMBO J., 3, pp.2555~2560 (1984)
  6. Liu S-Y, "Purification and cloning of a thermostable xylose (glucose) isomerase with an acidic pH optimum from Thermoanaerobacterium strain JW", SL-YS 489, 1996, pp.178, 5938~5945
  7. Kikuchi T, "Molecular cloning of the xylA gene encoding xylose isomerase from Streptomyces griseofuscus S-41: primary structure of the gene and its product", Agric. Biol. Chem., 54, pp.2469~2472 (1990)
  8. Kevbrin VV, "Anoxybacillus kamchatkensis sp. nov., a novel thermophilic facultative aerobic bacterium with a broad pH optimum from the Geyser valley, Kamchatka", Extremophiles, 9, pp.391~398 (2005)[DOI]
  9. Lee S-J, "Draft genome sequence of the thermophilic bacterium Anoxybacillus kamchatkensis G10", J. Bacteriol., 194, pp.6684~6685 (2012)[DOI]
  10. Cihan AC, "Taxonomic classification of Anoxybacillus isolates from geothermal regions in Turkey by 16S rRNA gene sequences and ARDRA, ITS-PCR, Rep-PCR analyses", Pol. J. Microbiol., 62, pp.149~163 (2013)
  11. Poli A, "Anoxybacillus amylolyticus sp. nov., a thermophilic amylase producing bacterium isolated from Mount Rittmann (Antarctica)", Syst. Appl. Microbiol., 29, pp.300~307 (2006)[DOI]
  12. Ertunga NS, "Cloning, expression, purification and characterization of fructose-1,6-bisphosphate aldolase from Anoxybacillus gonensis G2", J. Biochem., 141, pp.817~825 (2007)[DOI]
  13. Colak A, "Characterization of a thermoalkalophilic esterase from a novel thermophilic bacterium, Anoxybacillus gonensis G2", Bioresour. Technol., 96, pp.625~631 (2005)[DOI]
  14. Lavrenteva E, "The study of two alkaliphilic thermophile bacteria of the Anoxybacillus genus as producers of extracellular proteinase", Appl. Biochem. Microbiol., 45, pp.484~488 (2009)[DOI]
  15. Kacagan M, "Characterization of a xylanase from a thermophilic strain of Anoxybacillus pushchinoensis A8", Biologia, 63, pp.599~606 (2008)
  16. Bradford MM, "A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding", Anal. Biochem., 72, pp.248~254 (1976)[DOI]
  17. Dische Z, "A new spectrophotometric method for the detection and determination of keto sugars and trioses", J. Biol. Chem., 192, pp.583~587 (1951)
  18. Chauthaiwale J, "Production and purification of extracellular D-xylose isomerase from an alkaliphilic, thermophilic Bacillus sp.", Appl. Environ. Microbiol., 60, pp.4495~4499 (1994)
  19. Jari Oksanen, "Vegan: Community Ecology Package. R package version 2.4-4", ~ (2017)
  20. Lajoie CA, "Cloning, expression and characterization of xylose isomerase from the marine bacterium Fulvimarina pelagi in Escherichia coli", Biotechnol. Prog., 32, pp.1230~1237 (2016)[DOI]
  21. Karaoglu H, "Biochemical characterization of a novel glucose isomerase from Anoxybacillus gonensis <TEX>$G2^T$<", TEX> that displays a high level of activity and thermal stability, 2013, pp.97, 215~224
  22. Vieille C, "xylA cloning and sequencing and biochemical characterization of xylose isomerase from Thermotoga neapolitana", Appl. Environ. Microbiol., 61, pp.1867~1875 (1995)
  23. Hartley BS, "Glucose isomerase: insights into protein engineering for increased thermostability", Biochim. Biophys. Acta, 1543, pp.294~335 (2000)[DOI]
  24. Allen KN, "Role of the divalent metal ion in sugar binding, ring opening, and isomerization by <TEX>$\small{D}$<", TEX>-xylose isomerase: replacement of a catalytic metal by an amino acid, 1994, pp.33, 1488~1494
  25. Kovalevsky AY, "Metal ion roles and the movement of hydrogen during reaction catalyzed by <TEX>$\small{D}$<", TEX>-xylose isomerase: a joint X-ray and neutron diffraction study, 2010, pp.18, 688~699
  26. Callens M, "Catalytic properties of <TEX>$\small{D}$<", TEX>-xylose isomerase from Streptomyces violaceoruber, 1986, pp.8, 696~700
  27. Lehmacher A, "Comparative kinetics of <TEX>$\small{D}$<", TEX>-xylose and <TEX>$\small{D}$<, TEX>-xylose isomerase from Thermus aquaticus HB8, pp.Biol. Chem. Hoppe Seyler, 1990[DOI]
  28. Hlima HB, "Engineered glucose isomerase from Streptomyces sp. SK is resistant to <TEX>$Ca^{2+}$<", TEX> inhibition and <TEX>$Co^{2+}$<, J. Ind. Microbiol. Biotechnol., pp.2012, 39[DOI]
  29. Faiz O, "Cloning, expression and characterization of xylose isomerase from thermophilic Geobacillus caldoxylosilyticus TK4 strain", Turk. J. Biochem., 36, pp.6~14 (2011)
  30. Lama L, "Purification and characterization of thermostable xylose (glucose) isomerase from Bacillus thermoantarcticus", J. Ind. Microbiol. Biotechnol., 27, pp.234~240 (2001)[DOI]
  31. Brown SH, "Purification and characterization of a highly thermostable glucose isomerase produced by the extremely thermophilic eubacterium, Thermotoga maritima", Biotechnol. Bioeng., 41, pp.878~886 (1993)[DOI]
  32. Smith CA, "<TEX>$\small{D}$<", TEX>-Xylose (<TEX>$\small{D}$<, Biochem. J., pp.1991, 277[DOI]
  33. Joo G-J, "Purification and characterization of a thermostable xylose (glucose) isomerase from Streptomyces chibaensis J-59", Agric. Chem. Biotechnol., 44, pp.113~118 (2001)
  34. Jia DX, "Properties of a novel thermostable glucose isomerase mined from Thermus oshimai and its application to preparation of high fructose corn syrup", Enzyme Microb. Technol., 99, pp.1~8 (2017)[DOI]

관련 논문AI추천

Crystal Structure and Functional Characterization of a Xylose Isomerase (PbXI) from the Psychrophilic Soil Microorganism, Paenibacillus sp.권성학, 김경진, 김창민, 김학준, 박선하, 박현호, 이준혁, 이창우, 정창숙, 홍종욱한국미생물·생명공학회 · Journal of Microbiology and Biotechnology2019 · 제29권 제2호Electrostatic Immobilization of D-Xylose Isomerase to a Cation Exchanger for the Conversion of D-Xylose to D-Xylulose권대혁, 김성건, Hang, Nguyen Thi한국미생물·생명공학회 · 한국미생물·생명공학회지2012 · 제40권 제2호Purification and Characterization of a Thermostable Xylanase from Paenibacillus sp. NF1 and its Application in Xylooligosaccharides ProductionFuping Lu, Hai-sheng Pei, Hong-chen Zheng, Jing-sheng Zhang, Jin-rong Hu, Jun-she Sun, Ling-cai Meng, Ming-zhe Sun, Wen-hui Zeng, Xiu-Qing Zhang, Zheng Yan한국미생물·생명공학회 · Journal of Microbiology and Biotechnology2014 · 제24권 제4호Establishment of High-Throughput Screening Protocol Based on Isomerase Using Geobacillus sp. L-Rhamnose IsomeraseBaek Seung-Ho, Han Sol Min, Kim Seong-Bo, Koo Na Kyeong, Park Hyun June한국미생물·생명공학회 · Journal of Microbiology and Biotechnology2025 · 제35권 Molecular Characterization of a Thermophilic and Salt- and Alkaline-Tolerant Xylanase from Planococcus sp. SL4, a Strain Isolated from the Sediment of a Soda LakeGuozeng Wang, Juan Lin, Xiaoyun Huang, Xiuyun Ye한국미생물·생명공학회 · Journal of Microbiology and Biotechnology2015 · 제25권 제5호Lactulose Production Using Immobilized Cells Including Thermostable Cellobiose 2-epimerase구봉성, 김은정, 김진숙, 박아름, 이현철한국미생물·생명공학회 · 한국미생물·생명공학회지2016 · 제44권 제4호

이 간행물의 다른 논문 (9건)