Kinetic and thermodynamic study of cloned thermostable endo-1,4-β-xylanase from Thermotoga petrophila in mesophilic host

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• 作者：Ikram ul Haq (1) ikmhaq@yahoo.com
  Zahid Hussain (1) zahid_biotech@yahoo.com
  Mahmood Ali Khan (1)
  Bushra Muneer (1)
  Sumra Afzal (1)
  Sana Majeed (1)
  Fatima Akram (1)
• 关键词：Cloning – Hyperthermophilic – Endo ; 1 ; 4 ; β ; xylanase – Thermodynamics – Thermotoga petrophila
• 刊名：Molecular Biology Reports
• 出版年：2012
• 出版时间：July 2012
• 年：2012
• 卷：39
• 期：7
• 页码：7251-7261
• 全文大小：1.0 MB
• 参考文献：1. Owens B, Tucker L, Collins MA, McCracken KJ (2008) Effects of different feed additives alone or in combination on broiler performance, gut microflora and ileal histology. Br Poult Sci 49(2):202–212
  2. Zheng H, Guo B, Chen XL, Fan SJ, Zhang YZ (2011) Improvement of the quality of wheat bread by addition of glycoside hydrolase family 10 xylanases. Appl Microbiol Biotechnol 90(2):509–515
  3. Dwivedi P, Vivekanand V, Pareek N, Sharma A, Singh RP (2010) Bleach enhancement of mixed wood pulp by xylanase–laccase concoction derived through co-culture strategy. Appl Biochem Biotechnol 160(1):255–268
  4. Gibbs MD, Reeves RA, Bergquist PL (1995) Cloning, sequencing and expression of a xylanase gene from the extreme thermophile Dictyoglomus thermophilum Rt46B.1 and activity of the enzyme on fiber-bound substrate. Appl Environ Microbiol 61:4403–4408
  5. Sunna A, Antranikian G (1997) Growth and production of xylanolytic enzymes by the extreme thermophilic anaerobic bacterium Thermotoga thermarum. Appl Microbiol Biotechnol 45:671–676
  6. Simpson HD, Haufler UR, Daniel RM (1991) An extremely thermostable xylanase from the thermophilic eubacterium Thermotoga. Biochem J 277:177–185
  7. Winterhalter C, Liebl W (1995) Two extremely themostable xylanases of the hyperthermophilic bacterium Thermotoga maritima MSB8. Appl Environ Microbiol 61:1810–1815
  8. Bok JD, Goers SK, Eveleigh DE (1994) Cellulase and xylanase systems of Thermotoga neapolitana. ACS Symp Ser 566:54–65
  9. Sunna A, Puls J, Antranikian G (1996) Purification and characterization of two thermostable endo-1,4-β-d-xylanases from Thermotoga thermarum. Biotechnol Appl Biochem 24:177–185
  10. Yang JL, Eriksson KEL (1992) Use of hemicellulolytic enzymes as one stage in bleaching of kraft pulps. Holzforschung 46:481–488
  11. Chen CC, Adolphson R, Dean FDJ, Eriksson KEL, Adamas MWW, Westpheling J (1997) Release of lignin from kraft pulp by a hyperthermophilic xylanase from Thermotoga maritima. Enzyme Microbiol Technol 20:39–45
  12. Widdel F, Kohring G, Mayer F (1983) Studies in dissimilatory sulfate-reducing bacteria that decompose fatty acids. III. Characterization of the filamentous gliding Desulfonema limicola gen. nov. sp. Nov., and Desulfonema magnum sp. Nov. Arch Microbiol 134:286–294
  13. Kibbe WA (2007) OligoCalc: an online oligonucleotide properties calculator. Nucleic Acids Res 35(Suppl 2):W43–W46. doi:
  14. Sambrook J, Russell DW (2001) Molecular cloning: a laboratory manual. Cold Spring Harbor, New York
  15. Miller GL (1959) Use of dinitrosalicylic acid reagent for determination of reducing sugar. Anal Chem 31:426–428
  16. Bradford MM (1976) A dye binding assay for protein. Anal Biochem 72:248–254
  17. Bokhari SAI, Latif F, Rajoka MI (2009) Purification and characterization of xylanases from Thermomyces lanuginosus and its mutant derivative possessing novel kinetic and thermodynamic properties. World J Microbiol Biotechnol 25:493–502
  18. Henrissat B, Bairoch A (1993) New families in the classification of glycosyl hydrolases based on amino-acid sequence similarities. Biothem J 293:781–788
  19. Henrissat B, Romeo A (1995) Families, superfamilies. And subfamilies of glycosyl hydrolases. Biochem J 311:350–351
  20. Davies G, Hanrissat B (1995) Structures and mechanisms of glycosyl hydrolases. Structure 3(9):853–859
  21. Hemissat B, Callebaut I, Fabrega S, Lehn P, Momon JP, Davies G (1995) Conserved catalytic machinery and the prediction of a common fold for several families of glycosyl hydrolases. Proc Natl Acad Sci USA 92:7090–7094
  22. Ihsanawati KT, Kaneko T, Morokuma C, Yatsunami R, Sato T, Nakamura S, Tanaka N (2005) Structural basis of the substrate subsite and the highly thermal stability of xylanase 10B from Thermotoga maritima MSB8. Proteins 61:999–1009
  23. Santos CR, Meza AN, Hoffmam ZB, Silva JC, Alvarez TM, Ruller R, Giesel GM, Verli H, Squina FM, Prade RA, Murakami MT (2010) Thermal-induced conformational changes in the product release area drive the enzymatic activity of xylanases 10B: crystal structure, conformational stability and functional characterization of the xylanase 10B from Thermotoga petrophila RKU-1. Biochem Biophys Res Commun 403(2):214–219
  24. Turner P, Hoist O, Nordberg KE (2005) Optimized expression of soluble cyclomaltodextrinase of thermophilic origin in Escherichia coli by using a soluble fusion-tag and by tuning of inducer concentration. Protein Expr Purif 39:54–60
  25. Yemin X, Xianfei S, Jinjin Y (2009) Overexpression of β-glucosidase from Thermotoga maritima for the production of highly purified aglycone isoflavones from soy flour. World J Microbiol Biotechnol 25(12):2165–2172
  26. Brown SH, Sjoholm C, Kelly RM (1993) Purification and characterization of a highly thermostable glucose isomerase produced by the extremely thermophilic eubacterium, Thermotoga maritima. Biotechnol Bioeng 41:878–886
  27. Ruttersmith LD, Daniel RM (1991) Thermostable cellobiohydrolase from the thermophilic eubacterium Thermotoga sp. Strain FjSS3-B.l. Biochem J 277:887–890
  28. Mathrani IM, Ahring BK (1992) Xylanases from several Dictyoglomus isolates. In: Visser J, Beldman G, Kusters-van Someren MA, Voragen AGJ (eds) Xylans and xylanases. Elsevier, Amsterdam, pp 483–486
  29. Ruttersmith LD, Daniel RM, Simpson HD (1992) Cellulolytic and hemicellulolytic enzymes functional above 100°C. Ann N Y Acad Sci 672:137–141
  30. Dahlberg L, Holst O, Kristjansson JK (1993) Thermostable xylanolytic enzymes from Rhodothermus marinus grown on xylan. Appl Microbial Biotechnol 40:63–68
  31. Sunna A, Jiirgen P, Gumbed A (1997) Characterization of the xylanolytic enzyme system of the extreme thermophilic anaerobic bacteria Thermotoga maritima, T. neapolitana, and T. thermarum Comp. Biochem Physiol 118:453–461
  32. Wong KKY, Tan LUL, Saddler JN (1988) Multiplicity of β-1,4-xylanase in microorganisms: function and applications. Microbiol Rev 52:305–317
  33. Jiang ZQ, Deng W, Zhu YP, Li LT, Sheng YJ, Hayashi K (2004) The recombinant xylanase B of Thermotoga maritima is highly xylan specific and produces exclusively xylobiose from xylans, a unique character for industrial applications. J Mol Catalysis B: Enzymatic 27:207–213
  34. Sajjad M, Muhammad IMK, Nadeem SA, Sajjad A, Imran A, Muhammad WA (2010) Enhanced expression and activity yields of Clostridium thermocellum xylanases without non-catalytic domains. J Biotechnol 145:38–42
  35. Cambillau C, Claverie JM (2000) Structural and genomic correlates of hyperthermostability. J Biol Chem 275:32383–32386
  36. Suhre K, Claverie JM (2003) Genomic correlates of hyperthermostability, an update. J Biol Chem 278:17198–17202
  37. Benkovic SJ, Hammes-Schiffer SA (2003) A perspective on enzyme catalysis. Science 301:1196–1202
  38. Rajagopalan PT, Benkovic SJ (2002) Preorganization and protein dynamics in enzyme catalysis. Chem Records 2:24–36
  39. Bokhari SAI, Rajoka MI, Javed A, Shafiq ur Rehman, Ishtiaq ur Rehman, Latif F (2010) Novel thermodynamics of xylanase formation by a 2-deoxy-d-glucose resistant mutant of Thermomyces lanuginosus and its xylanase potential for biobleachability. Bioresour Technol 101:2800–2808
  40. Afzal AJ, Ali S, Latif F, Rajoka MI, Siddiqui KS (2005) Innovative kinetic and thermodynamic analysis of a purified superactive xylanase from Scopulariopsis sp. Appl Biochem Biotechnol 120:51–70
  41. Vieille C, Zeikus JG (1996) Thermozymes: identifying molecular determinants of protein structural and functional stability. Trends Biotechnol 14:183–190
  42. Chen J, Lu Z, Sakkon J, Sites WE (2000) Increasing the thermostability of staphylococcal nuclease: implications for the origin of protein thermostability. J Mol Biol 303:125–130
  43. Georis J, Esteves FL, Brasseur JL, Bougnet V, Devreese B, Giannotta F (2000) An additional aromatic interaction improves the thermostability and thermophilicity of a mesophilic family 11 xylanase: structural basis and molecular study. Protein Sci 9:466–475
• 作者单位：1. Institute of Industrial Biotechnology, GC University, Lahore, 54000 Pakistan
• 刊物类别：Biomedical and Life Sciences
• 刊物主题：Life Sciences
  Animal Anatomy, Morphology and Histology
  Animal Biochemistry
  
• 出版者：Springer Netherlands
• ISSN：1573-4978

文摘

The 1,044 bp endo-1,4-β-xylanase gene of a hyperthermophilic Eubacterium, “Thermotoga petrophila RKU 1” (T. petrophila) was amplified, from the genomic DNA of donor bacterium, cloned and expressed in mesophilic host E. coli strain BL21 Codon plus. The extracellular target protein was purified by heat treatment followed by anion and cation exchange column chromatography. The purified enzyme appeared as a single band, corresponding to molecular mass of 40 kDa, upon SDS-PAGE. The pH and temperature profile showed that enzyme was maximally active at 6.0 and 95°C, respectively against birchwood xylan as a substrate (2,600 U/mg). The enzyme also exhibited marked activity towards beech wood xylan (1,655 U/mg). However minor activity against CMC (61 U/mg) and β-Glucan barley (21 U/mg) was observed. No activity against Avicel, Starch, Laminarin and Whatman filter paper 42 was observed. The K _m , V _max and K _cat of the recombinant enzyme were found to be 3.5 mg ml?1, 2778 μmol mg?1min?1 and 2,137,346.15 s?1, respectively against birchwood xylan as a substrate. The recombinant enzyme was found very stable and exhibited half life (t _?) of 54.5 min even at temperature as high as 96°C, with enthalpy of denaturation (ΔH*_D), free energy of denaturation (ΔG*_D) and entropy of denaturation (ΔS*_D) of 513.23 kJ mol?1, 104.42 kJ mol?1 and 1.10 kJ mol?1K?1, respectively at 96°C. Further the enthalpy (ΔH*), Gibbs free energy (ΔG*) and entropy (ΔS*) for birchwood xylan hydrolysis by recombinant endo-1,4-β-xylanase were calculated at 95°C as 62.45 kJ mol?1, 46.18 kJ mol?1 and 44.2 J mol?1 K?1, respectively.