Abstract
A thermo stable xylanase was purified from Streptomyces thermocyaneoviolaceus M049 for the production of xylooligosaccharides from xylan. The enzyme showed thermostability by maintaining 65% of remaining enzyme activity after 1 h heat treatment at 70°C. The molecular weight of the purified protein was 35 kDa in SDS-PAGE, and the optimal pH and temperature for the enzymatic activity were pH 5.0 and 60°C, respectively. N-terminal amino acid sequences of the purified xylanase, DTITSNQTGTHNGYF, were similar to StxII from S. Thermoviolaceus and XlnB from S. lividans. Using those two genes, stxll and xlnB as probe DNA, a gene encoding xylanase, xynB, was cloned from genomic library of S. thermocyaneoviolaceus M049. The open reading frame of the xynB was composed of 1008 nucleotide sequences. Compared to N-terminal sequences from purified enzyme, it was proposed that the XynB contained a 40 amino acid long signal peptide to the N-terminus. For easy production and purification, a XynB overproduction strain was constructed using pET21a(+) and strain E. coli BLR(DE3). Consequently, the recombinant enzyme was tested for the production of xylooligosaccharides through TLC and HPLC analyses.
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Kulkarni, N., E. A. Shendy, and M. Rao (1999) Molecular and biotechnological aspects of xylanase. FEMS Microbiol. Rev. 23: 411–456.
Liab, K., P. Azadi, R. Collins, J. Tolan, J. Kim, and K. Eriksson (2000) Relationships between activities of xylanases and xylan structures. Enzym. Microb. Technol. 27: 89–94.
Biely, P. (1985) Microbial xylanolytic systems. Trends Biotechnol. 3: 286–290.
Jeya, M., S. Thiagarajan, Jung-Kul Lee, and P. Gunasekaran (2009) Identification of new GH 10 and GH 11 xylanase genes from Aspergillus versicolor MKU3 by genome-walking PCR. Biotechnol. Bioprocess Eng. 14: 13–19.
Dekker, R. F. H. and G. N. Richards (1976) Hemicellulases: their occurrence, purification, properties, and mode of action. Adv. Carbohydr. Chem. Biochem. 32: 277–352.
Frederick, M. M., C. H. Kiang, J. R. Frederick, and P. J. Reilly (1985) Purification and characterization of endoxylanases from Aspergillus niger. I. Two isozymes active on xylan backbones near branch points. Biotechnol. Bioeng. 27: 525–532.
Irwin, D., E. D. Jung, and D. B. Wilson (1994) Characterization and sequence of a Thermomonospora fusca xylanase. Appl. Environ. Microbiol. 60: 763–770.
Reilly, P. J. (1981) Xylanase: structure and function in trends in the biology of fermentations for fuels and chemicals. Basic Life Sci. 18: 111–129.
Wong, K. K. Y. and J. N. Saddler (1993) Applications of hemicellulases in the food, feed and pulp, and paper industries. Hemicellulose and Hemicellulases. pp. 127–143. Portland Press, London, UK.
Kim, K. C., S. W. Kim, M. J. Kim, and S. J. Kim (2005) Sacchrification of foodwastes using cellulolytic and amylolytic enzymes from Trichoderma harzianum FJ1 and its kinetics. Biotechnol. Bioprocess Eng. 10: 52–59.
Chen C., J. L. Chen, and T.Y. Lin (1997) Purification and characterization of a xylanase from Trichoderma longibrachiatum for xylooligosaccharide production. Enzym. Microb. Technol. 21: 91–96.
Jeong, K. J., I. Y. Park, M. S. Kim, and S. C. Kim (1998) High-level expression of an endoxylanase gene from Bacillus sp. in Bacillus subtilis DB104 for the production of xylobiose from xylan. Appl. Microbiol. Biotechnol. 50: 113–118.
Jiang, Q., W. Deng., Y. P. Zhu., L. T. Li., Y. J. Sheng, and K. Hayashi (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. Catal. B: Enzymatic 27: 207–213.
Vázquez, M. J., J. L. Alonso., H. Domínguez, and J. C. Parajó (2000) Xylooligosaccharides: manufacture and applications. Trends Food Sci. Technol. 11: 387–393.
Yuan, Q. P., H. Zhang, Z. M. Qian, and X. J. Yang (2004) Pilot-plant production of xylo-oligosaccharides from corncob by steaming, enzymatic hydrolysis, and nonofiltration. J. Chem. Technol. Biot. 79: 1073–1079.
Björkstén, B., E. Sepp, K. Julge, T. Voor, and M. Mikelsaar (2001) Allergy development and the intestinal microflora during the first year of life. J. Allergy and Clinical Immunology. 108: 516–520.
Guarner, F. and J. R. Malagelada (2003) Gut flora in health and disease. The Lancet 361: 512–519.
Basaran, P., Y. D. Hang, N. Bsaran, and R. W. Worobo (2001) Cloning and heterologous expression of xylanase from Pichia stipitis in Escherichia coli. J. Appl. Microbiol. 90: 248–255.
Cho, S. G. and Y. J. Choi (1995) Nucleotide sequence analysis of an endo-xylanase gene(xynA) from Bacillus stearothermophilus. J. Microbiol. Biotechnol. 5: 117–124.
Choi, J. H., O. S. Lee, J. H. Shin, Y. Y. Kwak, Y. M. Kim, and I. K. Rhee (2006) Thermostable xylanase encoded by xynA of Streptomyces thermocyaneoviolaceus: cloning, purification, characterization, and production of xylooligosaccharides. J. Microbiol. Biotechnol. 16: 57–63.
Kimura, T., H. Suzuki, H. Furuhashi, T. Aburatani, K. Morimoto, S. Karita, K. Sakka, and K. ohmiya (2000) Molecular cloning, overexpression, and purification of a major xylanase from Aspergillus oryzae. Biosci. Biotechnol. Biochem. 64: 2734–2738.
Srivastava, R., S. S. Ali, and B. S. Srivastara (1991) Cloning of xylanase gene of Streptomyces flavogriseus in Escherichia coli and becteriophage λ-induced lysis for the release of cloned enzyme. FEMS Microbiol. Lett. 78: 201–206.
Vieira, J. and J. Messing (1987) Production of singlestranded plasmid DNA. Methods Enzymol. 153: 3–11.
Bradford, M. M. (1976) A rapid and sensitive method for the quantitification of microgram quantities of protein utilizing the principle of protein dye binding. Anal. Biochem. 72: 248–254.
Jung Y. J., J. S. Yoo, Y. S. Lee, I. H. Park, S. H. Kim, S. C. Lee, M. Yasuda, S. Y. Chung, and Y. L. Choi (2007) Purification and characterization of thermostable β-1,3-1,4 glucanase from Bacillus sp. A8-8. Biotechnol. Bioprocess Eng. 12: 265–270.
Miller, G. L. (1959) Use of dinitrosalicylic acid reagent for determination of reducing sugar. Anal. Chem. 31: 426–428.
Hunkapiller, M. W. and L. E. Hood (1983) Analysis of phenylthiohydantoins by ultrasensitive gradient highperformance liquid chromatography. Methods Enzymol. 91: 486–493.
Sambrook, J., E. F. Fritch, and T. Maniatis (1989) Molecular cloning: A Laboratory Manual. 2nd ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA.
Hopwood, D. A., M. J. Bibb, K. F. Chater, T. Kieser, C. J. Bruton, H. M. Kieser, D. J. Lydiate, C. P. Smith, J. M. Ward, and H. Shrempf (1985) Genetic Manipulation of Streptomyces: A Laboratory Manual. The John Innes Foundation, Norwich, England.
Sanger, F., S. Nicklen, and A. R. Coulson (1977) DNA sequencing with chain — terminating inhibitors. Proc. Natl. Acad. Sci. 74: 5463–5467.
Tsujibo, H., K. Miyamoto, T. Kuda, K. Minami, T. Sakamoto, R. Hasegawa, and Y. Inamori (1992) Purification, properties and partial amino acid sequence of thermostable xylanases Streptomyces thermoviolaceus OPC-520. Appl. Environ. Microbiol. 58: 371–375.
Henrissat, B. and A. Bairoch (1993) Updating the sequence-based classification of glycosyl hydrolases. Biochem. J. 316: 695–696.
Tsujibo, H., T. Ohtsuki, T. Iio, I. Yamazaki, K. Miyamoto, M. Sugiyama, and Y. Inamori (1997) Cloning and sequence analysis of genes encoding xylanases and acetyl xylan esterase from Streptomyces thermoviolaceus OPC-520. Appl. Environ. Microbiol. 63: 661–664.
Lee, O. S., C. S. Choi, J. H. Choi, G. J. Joo, and I. K. Rhee (2001) Production of xylooligosaccharides with thermostable xylanase from Streptomyces thermocyaneoviolaceus. Kor. J. Appl. Microbiol. Biotechnol. 29: 221–226.
Aachary, A. A. and S. G. Prapulla (2008) Corncobinduced endo-1,4-β-D-xylanase of Aspergillus oryzae MTCC 5154: Production and characterization of xylobiose from glucuronoxylan. J. Agric. Food Chem. 56: 3981–3988.
Akpinar, O., O. Ak, A. Kavas, U. Bakir, and L. Yilmaz (2007) Enzymatic production of xylooligosaccharides from cotton stalks. J. Agric. Food Chem. 55: 5544–5551.
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Shin, JH., Choi, JH., Lee, OS. et al. Thermostable xylanase from Streptomyces thermocyaneoviolaceus for optimal production of xylooligosaccharides. Biotechnol Bioproc E 14, 391–399 (2009). https://doi.org/10.1007/s12257-008-0220-3
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DOI: https://doi.org/10.1007/s12257-008-0220-3