A thermostable xylanase was purified and characterized from the thermophilic fungus Rhizomucor miehei (Cooney & Emerson) Schipper. The enzyme was purified to homogeneity by ammonium sulfate precipitation, sephadex G-100 gel filtration and diethylaminoethyl cellulose anion exchange chromatography with a 29.1-fold. The enzyme was highly active within a range of pH from 5.0 to 6.5. The optimum temperature of the purified enzyme was 75 °C. The enzyme showed high thermal stability at 70 °C and 75 °C and the half-life of the xylanase at 90 °C was 30 min. Km and Vmax values at 50 °C of the purified enzyme were 0.055 mg/ml and 113.5 μmol min−1 mg−1, respectively. The enzyme was activated by Ca2+, Cu2+, K+ and Na+. On the other hand, Ag2+, Hg2+, Ba2+, and Zn2+ inhibited the enzyme. The molecular weight of the xylanase was estimated to be 27 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The present study is among the first works to examine and describe a secreted highly thermostable endoxylanase from the Rhizomucor miehei fungus. This enzyme displays a number of biochemical properties that make it a potentially strong candidate for industrial and commercial application in pulp bleaching.
Anand, L., Krishnamurthy, S., Vithayathil, P. J. (1990) Purification and properties of xylanase from the thermophilic fungus, Humicola lanuginosa Griffon and Maublanc Bunce. Arch. Biochem. Biophys. 276, 546–553.
Vithayathil P. J. , 'Purification and properties of xylanase from the thermophilic fungus, Humicola lanuginosa Griffon and Maublanc Bunce ' (1990 ) 276 Arch. Biochem. Biophys. : 546 -553.
Annamalai, N., Thavasi, R., Jayalakshmi, S., Balasubramanian, T. (2009) Thermostable and alkaline tolerant xylanase production by Bacillus subtilis isolated form marine environment. Ind. J. Biotechnol. 8, 291–297.
Balasubramanian T. , 'Thermostable and alkaline tolerant xylanase production by Bacillus subtilis isolated form marine environment ' (2009 ) 8 Ind. J. Biotechnol. : 291 -297.
Archana, A., Sharma, A., Satyanarayana, T. (1997) Xylanolytic enzymes. In: Johri, B. N., Satyanarayana, T., Olsen, J. (eds) Thermophilic Molds in Biotechnology. Kluwer Academic, pp. 169–190.
Bailey, M. J., Biely, P., Poutanen, K. (1992) Interlaboratory testing of methods for assay of xylanase activity. J. Biotechnol. 23, 257–270.
Poutanen K. , 'Interlaboratory testing of methods for assay of xylanase activity ' (1992 ) 23 J. Biotechnol. : 257 -270.
Beg, Q. K., Kapoor, M., Mahajan, L., Hoondal, G. S. (2001) Microbial xylanase and their industrial application: a review. Appl. Microbiol. Biotechnol. 56, 326–338.
Hoondal G. S. , 'Microbial xylanase and their industrial application: a review ' (2001 ) 56 Appl. Microbiol. Biotechnol. : 326 -338.
Bennett, N. A., Ryan, J., Biely, P., Vrsanska, L., Kremnicky, L., Macris, B. J., Kekos, D., Christakopoulos, P., Katapodis, P., Claeyssens, M., Nerinckx, W., Ntauma, P., Bhat, M. K. (1998) Biochemical and catalytic properties of an endoxylanase purified from the culture filtrate of Thermomyces lanuginosus ATCC 46882. Carbohydr. Res. 306, 445–455.
Bhat M. K. , 'Biochemical and catalytic properties of an endoxylanase purified from the culture filtrate of Thermomyces lanuginosus ATCC 46882 ' (1998 ) 306 Carbohydr. Res. : 445 -455.
Bradford, M. A. (1976) A rapid and sensitive method for quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72, 248–254.
Bradford M. A. , 'A rapid and sensitive method for quantitation of microgram quantities of protein utilizing the principle of protein-dye binding ' (1976 ) 72 Anal. Biochem. : 248 -254.
Cesar, T., Mrša, V. (1996) Purification and properties of the xylanase produced by Thermomyces lanuginosus. Enzyme Microb. Technol. 19, 289–296.
Mrša V. , 'Purification and properties of the xylanase produced by Thermomyces lanuginosus ' (1996 ) 19 Enzyme Microb. Technol. : 289 -296.
Düsterhoft, E. M., Linssen, V. A. J. M., Voragen, A. G. J., Beldman, G. (1997) Purification, characterization, and properties of two xylanases from Humicola insolens. Enzyme Microb. Technol. 20, 437–445.
Beldman G. , 'Purification, characterization, and properties of two xylanases from Humicola insolens ' (1997 ) 20 Enzyme Microb. Technol. : 437 -445.
Ganju, R. K., Vithayathil, P. J., Murthy, S. K. (1989) Purification and characterization of two xylanase from Chaetomium thermophile var.coprophile. Can. J. Microbiol. 35, 836–842.
Murthy S. K. , 'Purification and characterization of two xylanase from Chaetomium thermophile var.coprophile ' (1989 ) 35 Can. J. Microbiol. : 836 -842.
Gomes, D. J., Gomes, J., Steiner, W. (1994) Production of highly thermostable xylanase by a wild strain of thermophilic fungus Thermoascus aurantiacus and partial characterization of the enzyme. J. Biotechnol. 37, 11–22.
Steiner W. , 'Production of highly thermostable xylanase by a wild strain of thermophilic fungus Thermoascus aurantiacus and partial characterization of the enzyme ' (1994 ) 37 J. Biotechnol. : 11 -22.
Haltrich, D., Nidetzky, B., Kulbe, K. D., Steiner, W., Zupaneie, S. (1996) Production of fungal xylanases. Bioresour. Technol. 58, 137–161.
Zupaneie S. , 'Production of fungal xylanases ' (1996 ) 58 Bioresour. Technol. : 137 -161.
Hayat, K., Nawaz, H., Latif, F., Asghar, M. (2001) Kinetics of Cellulase and xylanase of Chaetomium thermophile with respect to Aeration. Pakistan J. Biol. Sci. 47, 875–876.
Asghar M. , 'Kinetics of Cellulase and xylanase of Chaetomium thermophile with respect to Aeration ' (2001 ) 47 Pakistan J. Biol. Sci. : 875 -876.
Jiang, Z., Cong, Q., Yan, Q., Kumar, N., Du, X. (2010) Characterisation of a thermostable xylanase from Chaetomium sp. and its application in Chinese steamed bread. Food Chem. 120, 457–462.
Du X. , 'Characterisation of a thermostable xylanase from Chaetomium sp. and its application in Chinese steamed bread ' (2010 ) 120 Food Chem. : 457 -462.
Johri, B. N., Rawat, S. (2004) Xylanases of thermophilic molds and their application potential. In: Dilip, K. Arora (ed.) Handbook of Fungal Biotechnology. Chapter. 23. British Mycological Society, Cambridge University Press.
Khandke, K. M., Vithayathil, P. J., Murthy, S. K. (1989) Purification of xylanase, β-glucosidase, endocellulase and exocellulase from a thermophilic fungus Thermoascus aurantiacus. Arch. Biochem. Biophys. 274, 491–500.
Murthy S. K. , 'Purification of xylanase, β-glucosidase, endocellulase and exocellulase from a thermophilic fungus Thermoascus aurantiacus ' (1989 ) 274 Arch. Biochem. Biophys. : 491 -500.
Kitpreechavanich, V., Hayashi, M., Nagai, S. (1984) Purification and properties of endo-1,4-β-xylanase from Humicola lanuginosa. J. Ferment. Technol. 5, 415–420.
Nagai S. , 'Purification and properties of endo-1,4-β-xylanase from Humicola lanuginosa ' (1984 ) 5 J. Ferment. Technol. : 415 -420.
Kulkarni, N., Rao, M. (1999) Molecular and biotechnological aspects of xylanases. FEMS Microbiol. Reviews 23, 411–456.
Rao M. , 'Molecular and biotechnological aspects of xylanases ' (1999 ) 23 FEMS Microbiol. Reviews : 411 -456.
Laemmli, U. K. (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227, 680–685.
Laemmli U. K. , 'Cleavage of structural proteins during the assembly of the head of bacteriophage T4 ' (1970 ) 227 Nature : 680 -685.
Lineweaver, H., Burk, D. (1934) The determination of enzyme dissociation constants. J. American Chem. Society 56, 658–666.
Burk D. , 'The determination of enzyme dissociation constants ' (1934 ) 56 J. American Chem. Society : 658 -666.
Maalej, I., Belhaj, I., Masmoudi, N. F., Belghith, H. (2009) Highly thermostable xylanase of the thermophilic fungus Talaromyces thermophilus: Purification and characterization. Appl. Biochem. Biotechnol. 1581, 200–212.
Belghith H. , 'Highly thermostable xylanase of the thermophilic fungus Talaromyces thermophilus: Purification and characterization ' (2009 ) 1581 Appl. Biochem. Biotechnol. : 200 -212.
Maheshwari, R., Bharadwaj, G., Bhat, M. K. (2000) Thermophilic fungi: Their physiology and enzymes. Microbiol. Molec. Biol. Rev. 64, 461–488.
Bhat M. K. , 'Thermophilic fungi: Their physiology and enzymes ' (2000 ) 64 Microbiol. Molec. Biol. Rev. : 461 -488.
Mandels, M., Sternburg, D. (1976) Recent advances in cellular technology. J. Ferment. Technol. 54, 267–286.
Sternburg D. , 'Recent advances in cellular technology ' (1976 ) 54 J. Ferment. Technol. : 267 -286.
Miller, G. L. (1959) Use of dinitrosalicyclic reagent for the determination of reducing sugars. Anal. Chem. 31, 426–428.
Miller G. L. , 'Use of dinitrosalicyclic reagent for the determination of reducing sugars ' (1959 ) 31 Anal. Chem. : 426 -428.
Monti, R., Terenzi, H. F., Jorge, J. A. (1991) Purification and properties of an extracellular xylanase from the thermophilic fungus Humicola grisea var. thermoidea. Can. J. Microbiol. 37, 675–681.
Jorge J. A. , 'Purification and properties of an extracellular xylanase from the thermophilic fungus Humicola grisea var. thermoidea ' (1991 ) 37 Can. J. Microbiol. : 675 -681.
Palmer, T. (1991) Extraction and purification of enzymes. In: Palmer, T. (ed.) Understanding Enzymes. Ellis Horwood Ltd, England, pp. 301–317.
Palmer T. , '', in Understanding Enzymes , (1991 ) -.
Peterson, E. A., Sober, H. A. (1962) Column chromatography of protein: substituted cellulases. In: Colowich, S., Kapllan, N. (eds) Methods in Enzymology. Vol. 5. New York, pp. 3–27.
Plummer, D. T. (1978) The practice of column chromatography. In: Palmer, T. (ed.) An Introduction to Practical Biochemistry. McGraw-Hill Book Company “UK” Ltd, pp. 61–66.
Prabhu, K. A., Maheshwari, R. (1999) Biochemical properties of xylanases from a thermophilic fungus, Melanocarpus albomyces, and their action on plant cell walls. J. Biosci. 24, 461–470.
Maheshwari R. , 'Biochemical properties of xylanases from a thermophilic fungus, Melanocarpus albomyces, and their action on plant cell walls ' (1999 ) 24 J. Biosci. : 461 -470.
Singh, S., Madlala, A. M., Prior, B. A. (2003) Thermomyces lanyginosus: properties of strains and their hemicellulases. FEMS Microbial. Rev. 27, 3–16.
Prior B. A. , 'Thermomyces lanyginosus: properties of strains and their hemicellulases ' (2003 ) 27 FEMS Microbial. Rev. : 3 -16.
Tuohy, M. G., Coughlan, M. P. (1992) Production of thermostable xylan-degrading enzymes by Talaromyces emersonii. Biores. Technol. 39, 131–137.
Coughlan M. P. , 'Production of thermostable xylan-degrading enzymes by Talaromyces emersonii ' (1992 ) 39 Biores. Technol. : 131 -137.
Tuohy, M. G., Puls, J., Claeyssens, M., Vrsanska, M., Coughlan, M. P. (1993) The xylan-degrading enzyme system of Talaromyces emersonii: novel enzymes with activity against aryl β-D-xylosides and unsubstituted xylans. Biochem. J. 290, 515–523.
Coughlan M. P. , 'The xylan-degrading enzyme system of Talaromyces emersonii: novel enzymes with activity against aryl β-D-xylosides and unsubstituted xylans ' (1993 ) 290 Biochem. J. : 515 -523.
Wong, K. K. Y., Tan, L. U. L., Saddler, J. N. (1988) Multiplicity of β-1,4 xylanase in microorganisms’ functions and applications. Microbiol. Rev. 52, 305–317.
Saddler J. N. , 'Multiplicity of β-1,4 xylanase in microorganisms’ functions and applications ' (1988 ) 52 Microbiol. Rev. : 305 -317.