Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
권호 표지

Cloning of a Bacillus subtilis WL-7 Mannanase Gene and Characterization of the Gene Product

  • KWEUN , MIN-A (School of Food Science and Biotechnology, Woosong University) ;
  • LEE, MI-SUNG (School of Food Science and Biotechnology, Woosong University) ;
  • CHOI, JOON-HO (R&D Center, KoBioTech Co.) ;
  • CHO, KI-HAENG (R&D Center, CTCBIO Inc.) ;
  • YOON, KI-HONG (School of Food Science and Biotechnology, Woosong University, Bioresouce and Application Research Center, Woosong University)
  • Published : 2004.12.01
Download PDF
⟨ Previous Next ⟩

Abstract

A gene encoding the mannanase of Bacillus subtilis WL-7, which had been isolated from Korean soybean paste, was cloned into Escherichia coli, and the gene product was purified from the culture filtrate of the recombinant E. coli. This mannanase gene, designated manA, consisted of 1,086 nucleotides, encoding a polypeptide of 362 amino acid residues. The deduced amino acid sequence was highly homologous to those of mannanases belonging to the glycosyl hydrolase family 26. The molecular mass of the purified mannanase was 38 kDa as estimated by SDS-PAGE. The enzyme had a pH optimum at 6.0 and a temperature optimum at $55^{\circ}C$. The enzyme was active on locust bean gum, konjak, guar gum, and lichenan, while it did not exhibit activity towards yeast mannan, laminarin, carboxymethylcellulose, $\beta$­glucan, xylan, and para-nitrophenyl-$\beta$-mannopyranoside.

Keywords

References

  1. Ademark, P., A. Varga, J. Medve, V. Harjunpaa, T. Drakenberg, F. Tjerneld, and H. Stalbrand. 1998. Softwood hemicellulose-degrading enzymes from Aspergillus niger: Purification and properties of a $\beta$-mannanase. J. Biotechnol. 63: 199-210.
  2. Akino, T., C. Kato, and K. Horikoshi. 1989. The cloned $\beta$-mannanase gene from alkalophilic Bacillus sp. AM-001 produced two$\beta$-mannanases in Escherichia coli. Arch. Microbiol. 152: 10-15
  3. Altschul, S. F., W. Gish, W. Miller, E. W. Myers, and D. J. Lipman. Basic local alignment search tool. J. Mol. Biol. 215: 403-410
  4. Araujo, J. and O. P. Ward. 1990. Production, purification and properties of the mannanase components of Bacillus pumilus. Appl. Environ. Microbiol. 56: 1954-1956
  5. Arcand, N., D. Kluepfel, F. W. Paradis, R. Morosoli, and F. Shareck. 1993. $\beta$-Mannanase of Streptomyces lividans 66: Cloning and DNA sequence of the manA gene and characterization of the enzyme. Biochem. J. 290: 857-863
  6. Bradford, M. M. 1976. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72: 248- 254 https://doi.org/10.1016/0003-2697(76)90527-3
  7. Cann, I. K., S. Kocherginskaya, M. R. King, B. A. White, and R. I. Mackie. 1999. Molecular cloning, sequencing, and expression of a novel multidomain mannanase gene from Thermoanaerobacterium polysaccharolyticum. J. Bacteriol. 181: 1643-1651
  8. Christgau, S., S. Kauppinen, J. Vind, L.-V. Koffod, and H. Dalboge. 1994. Expression cloning, purification and characterization of a $\beta$-1,4-mannanase from Aspergillus aculeatus. Biochem. Mol. Biol. Int. 33: 917-925
  9. Duffaud G. D., C. M. McCutchen, P. Leduc, K. N. Parker, and R. M. Kelly. 1997. Purification and characterization of extremely thermostable $\beta$-mannanase, $\beta$-mannosidase, and $\alpha$-galactosidase from the hyperthermophilic eubacterium Thermotoga neapolitana 5068. Appl. Environ. Microbiol. 63: 169-177.
  10. Fanutti, C., T. Ponyi, G. W. Black, G. P. Hazlewood, and H. J. Gilbert. 1995. The conserved noncatalytic 40-residue sequence in cellulases and hemicellulases from anaerobic fungi functions as a protein docking domain. J. Biol. Chem. 270: 29314-29322
  11. Gibbs, M. D., A. U. Elinder, R. A. Reeves, and P. L. Bergquist. 1996. Sequencing, cloning and expression of a$\beta$-1,4- mannanase gene, manA, from the extremely thermophilic anaerobic bacterium, Caldicellulosiruptor Rt8B.4. FEMS Microbiol. Lett. 141: 37-43
  12. Gibbs, M. D., R. A. Reeves, A. Sunna, and P. L. Bergquist. 1999. Sequencing and expression of a $\beta$-mannanase gene from the extreme thermophile Dictyoglomus thermophilum Rt46B.1, and characteristics of the recombinant enzyme. Curr. Microbiol. 39: 351-357
  13. Grosjean, H. and W. Fiers. 1982. Preferential codon usage in genes: The optimal codon-anticodon interaction energyand the selective codon usage in efficiently expressed genes. Gene 18: 199-209
  14. Hagglund, P., T. Eriksson, A. Collen, W. Nerinckx, M. Claeyssens, and H. Stalbrand. 2003. A cellulose-binding module of the Trichoderma reesei $\beta$-mannanase Man5A increases the mannan-hydrolysis of complex substrates. J. Biotechnol. 101: 37-48
  15. Hogg, D., G. Pell, P. Dupree, F. Goubet, S. M. Martin-Orue, S. Armand, and H. J. Gilbert. 2003. The modular architecture of Cellvibrio japonicus mannanases in glycoside hydrolase families 5 and 26 points to differences in their role in mannan degradation. Biochem. J. 371: 1027-1043
  16. Johnvesly, B., S. Virupakshi, G. N. Patil, Ramalingam, and G. R. Naik. 2002. Cellulase-free thermostable alkaline xylanase from thermophilic and alkalophilic Bacillus sp. JB-99. J. Microbiol. Biotechnol. 12: 153-156
  17. Kansoh, A. L. and Z. A. Nagieb. 2004. Xylanase and mannanase enzymes from Streptomyces galbusNR and their use in biobleaching of softwood kraft pulp. Antonie van Leeuwenhoek 85: 103-114
  18. Kataoka, N. and Y. Tokiwa. 1998. Isolation and characterization of an active mannanase-producing anaerobic bacterium, Clostridium tertium KT-5A, from lotus soil. J. Appl. Microbiol. 84: 357-367 https://doi.org/10.1046/j.1365-2672.1998.00349.x
  19. Khanongnuch, C., T. Ooi, and S. Kinoshita. 1999. Cloning and nucleotide sequence of $\beta$-mannanse and cellulase genes from Bacillus sp. 5H. Wor. J. Microbiol. Biotechnol. 15: 249-258
  20. Kim, K. C., S.-S. Yoo, Y.-A. Oh, and S.-J. Kim. 2003. Isolation and characteristics of Trichoderma harzianum FJ1 producing cellulases and xylanase. J. Microbiol. Biotechnol. 13: 1-8
  21. Kim, M. D., K.-N. Kim, and Y. J. Choi. 2003. Enhanced expression of $\beta$-xylosidase of Bacillus stearothermophilus No. 236 by change of translational initiation codon in Escherichia coli and Bacillus subtilis. J. Microbiol. Biotechnol. 13: 584-590
  22. Kim, S. W., D. A. Knabe, K. J. Hong, and R. A. Easter. 2003. Use of carbohydrases in corn-soybean meal-based nursery diets. J. Anim. Sci. 81: 2496-2504
  23. Kim, Y.-H., M.-N. Gwon, S.-Y. Yang, T.-K. Park, C.-G. Kim, C.-W. Kim, and M.-D. Song. 2002. Isolation of phytaseproducing Pseudomonas sp. and optimization of its phytase production. J. Microbiol. Biotechnol. 12: 279-285
  24. Kurokawa, J., E. Hemjinda, T. Arai, S. Karita, T. Kimura, K. Sakka, and K. Ohmiya. 2001. Sequence of the Clostridium thermocellum mannanase gene man26B and characterization of the translated product. Biosci. Biotechnol. Biochem. 65: 548-554
  25. Kweun, M. A., H. S. Kim, M.-S. Lee, J. H. Choi, and K.-H. Yoon. 2003. Mannanase production by a soybean isolate, Bacillus subtilis WL-7. Kor. J. Microbiol. Biotechnol. 31: 277-283
  26. Mendoza, N. S., M. Arai, K. Sugimoto, M. Ueda, T. Kawaguchi, and L. M. Joson. 1995. Cloning and sequencing of $\beta$-mannanase gene from Bacillus subtilis NM-39. Biochim. Biophys. Acta 1243: 552-554.
  27. Mendoza, N. S., M. Arai, T. Kawaguchi, T. Yoshida, and L. M. Joson. 1994. Purification and properties of mannanase from Bacillus subtilis. Wor. J. Microbiol. Biotechnol. 10: 551-556
  28. Miller, M. L., R. Blum, W. E. Glennon, and A. L. Burton. 1960. Measurement of carboxymethylcellulase activity. Anal. Biochem. 2: 127-132
  29. Morris, D. D., R. A. Reeves, M. D. Gibbs, D. J. Saul, and P. L. Bergquist. 1995. Correction of the $\beta$-mannanase domain of the celC pseudogene from Caldocellulosiruptor saccharolyticus and activity of the gene product on kraft pulp. Appl. Environ. Microbiol. 61: 2262-2269
  30. Montiel, M. D., M. Hernandez, J. Rodriguez, and M. E. Arias. 2002. Evaluation of an endo-$\beta$-mannanase produced by Streptomyces ipomoea CECT 3341 for the biobleaching of pine kraft pulps. Appl. Microbiol. Biotechnol. 58: 67-72
  31. Oda, Y., T. Komaki, and K. Tonomura. 1993. Purification and properties of extracellular $\beta$-mannanases produced by Enterococcus casseliflavus FL2121 isolated from decayed konjac. J. Ferment. Bioeng. 76: 14-18
  32. Parker, K. N., S. R. Chhabra, D. Lam, W. Callen, G. D. Duffaud, M. A. Snead, J. M. Short, E. J. Mathur, and R. M. Kelly. 2001. Galactomannanases Man2 and Man5 from Thermotoga species: growth physiology on galactomannans, gene sequence analysis, and biochemical properties of recombinant enzymes. Biotechnol. Bioeng. 75: 322-333
  33. Politz, O., M. Krah, K. K. Thomsen, and R. Borriss. 2000. A highly thermostable endo-(1,4)-$\beta$-mannanase from the marine bacterium Rhodothermus marinus. Appl. Microbiol. Biotechnol. 53: 715-721
  34. Puls, J. and J. Schuseil. 1996. Chemistry of hemicelluloses: Relationship between hemicellulose structure and enzymes required for hydrolysis, pp. 1-28. In M. P. Coughlan and G. P. Hazlewood (eds.). Hemicelluloses and Hemicellulases. Portland Press, London, England
  35. Rodriquez, R. L. and R. C. Tait. 1983. Recombinant DNA Techniques - An Introduction, pp. 45-46. Addison-Wesley Pub. Co., Reading, MA, U.S.A
  36. Sadaie, Y., K. Yata, M. Fujita, H. Sagai, M. Itaya, Y. Kosahara, and N. Ogasawara. 1997. Nucleotide sequence and analysis of the phoB-rrnE-groESL region of the Bacillus subtilis chromosome. Microbiology 143: 1861-1866
  37. Sambrook, J., E. F. Fritsch, and T. Maniatis. 1989. Molecular Cloning. A Laboratory Mannual. 2nd ed. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, U.S.A
  38. Sharp, P. M., D. G. Higgins, D. C. Shields, and K. M. Devine. 1990. Protein-coding genes: DNA sequence database and codon usage, pp. 557-569. In C. R. Harwood and S. M. Cutting (eds.). Molecular Biological Methods for Bacillus. John Wiley & Sons, Chichester, England
  39. Stoll, D., A. Boraston, H. Stalbrand, B. W. McLean, D. G. Kilburn, and R. A. J. Warren. 2000. Mannanase Man26A from Cellulomonas fimi has a mannan-binding module. FEMS Microbiol. Lett. 183: 265-269
  40. Stoll, D., H. Stalbrand, and R. A. Warren. 1999. Mannandegrading enzymes from Cellulomonas fimi. Appl. Environ. Microbiol. 65: 2598-2605
  41. Sunna, A., M. D. Gibbs, C. W. J. Chin, P. J. Nelson, and P. L. Bergquist. 1999. A gene encoding a novel multidomain $\beta$-1,4-mannanase from Caldibacillus cellulovorans and action of the recombinant enzyme on kraft pulp. Appl. Environ. Microbiol. 66: 664-670
  42. Talbot, G. and J. Sygusch. 1990. Purification and characterization of thermostable $\beta$-mannanase and $\alpha$-galactosidase from Bacillus stearothermophilus. Appl. Environ. Microbiol. 56: 3505-3510
  43. Tamaru, Y., T. Araki, H. Amagoi, H. Mori, and T. Morishita. 1995. Purification and characterization of an extracellular $\beta$-1,4-mannanase from a marine bacterium, Vibrio sp. strain MA-138. Appl. Environ. Microbiol. 61: 4454-4458
  44. Tamaru, Y. and R. H. Doi. 2000. The engL gene cluster of Clostridium cellulovorans contains a gene for cellulosomal ManA. J. Bacteriol. 182: 244-247
  45. Valentine, P. J. and A. A. Salyers. 1992. Analysis of proteins associated with growth of Bacteroides ovatus on the branched galactomannan guar gum. Appl. Environ. Microbiol. 58: 1534-1540
  46. Yamaura, L., T. Matsumoto, M. Funatsu, and Y. Funatsu. 1990. Purification and some properties of endo-1,4-$\beta$-Dmannanase from Pseudomonas sp. PT-5. Agric. Biol. Chem. 54: 2425-2427