Summary
The thermophilic fungus Humicola grisea var. thermoidea produces a mycelium-associated β-xylosidase activity when grown in liquid-state cultures on media containing oat spelt xylan as the carbon source. The β-xylosidase was purified to apparent homogeneity by gel filtration and anion exchange chromatography. Its molecular weight was 37 and 50 kDa, as determined by MALDI/TOF mass spectrometry and SDS-PAGE, respectively. The purified enzyme exhibited maximum activity at 55 °C and pH 6.5. It was also active at pH 8.8, retaining 60% of its activity after 6 h of incubation at 50 °C. β-xylosidase was strongly inactivated by NBS and slightly activated by DTT and β-mercaptoethanol. The enzyme was highly specific for PNPX as the substrate. The purified β-xylosidase showed K m and V max values of 1.37 mM and 12.98 IU ml−1, respectively.
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Almeida E.M., Polizeli M.L.T.M., Terenzi H.F., Jorge J.A., 1995 Purification and biochemical characterization of a β-xylosidase from Humicola grisea var. thermoidea FEMS Microbiology Letters. 130:171–176
Blum H., Beier H., Gross B., 1987 Improved silver staining of plant proteins, RNA and DNA in polyacrylamide gels Electrophoresis 8:93–99
Collins T., Gerday C., Feller G., 2005 Xylanases, xylanase families and extremophilic xylanases FEMS Microbiology Reviews 29:3–23
Ferreira H.M.F., Filho E.X.F., 2004 Purification and characterization of a β-mannanase from Trichoderma harzianum strain T4 Carbohydrate Polymers 57:23–29
Filho E.X.F., 1996 Purification and characterization of a β-glucosidase from solid-state cultures of Humicola grisea var. thermoidea Canadian Journal of Microbiology 42:1–5
Herrmann M.C., Vrsanska M., Jurickckova M., Hirsch J., Biely P., Kubicek C.P., 1997 The β-xylosidase of Trichoderma reesei is a multifunctional β-d-xylan xylohydrolase Biochemical Journal 321:375–381
Kumar S., Ramon D., 1996 Purification and regulation of the synthesis of a β-xylosidase from Aspergillus nidulans FEMS Microbiology Letters 135:287–293
Laemmli U.K., (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4 Nature 227:680–685
Lama L., Calandrelli V., Gambacorta A., Nicolaus B., 2004 Purification and characterization of thermostable xylanase and β-xylosidase by thermophilic bacterium Bacillus thermantarcticus Research in Microbiology 155: 283–289
Leatherbarrow R.J., 1987 Enzfitter Manual. Biosoft, London, 13–42
Li Y.-K., Yao H.-J., Cho, Y.-tzu., 2000 Effective induction, purification and characterization of Trichoderma koningii G-39-xylosidase with high transferase activity Biotechnology and Applied Biochemistry 31:119–125
Lucena-Neto A.S., Filho E.X.F., 2004 Purification and characterization of a new xylanase from Humicola grisea var. thermoidea Brazilian Journal of Microbiology 35:86–90
Medeiros R.G., Hanada R., Filho E.X.F., 2003 Production of xylan-degrading enzymes from Amazon forest fungal species International Biodeterioration and Biodegradation 52:97–100
Pinphanichakarn P., Tangsakul T., Thongnumwon T., Talawanich Y., Thamchaipenet A., 2004 Purification and characterization of β-xylosidase from Streptomyces sp. CH7 and its gene sequence analysis World Journal of Microbiology and Biotechnology 20:727–733
Ramsom R.F., Walton J.D., 1997 Purification and characterization of extracellular β-xylosidase and α-arabinosidase from the plant pathogenic fungus Cochliobolus carbonum Carbohydrate Research 297:357–364
Saha B.C., 2001 Purification and characterization of an extracellular β-xylosidase from a newly isolated Fusarium verticillioides Journal of Industrial Microbiology and Biotechnology 27:241–245
Saha B.C., 2003 Purification and properties of an extracellular β-xylosidase from a newly isolated Fusarium proliferatum Bioresource Technology 90:33–38
Salles B.C., Cunha R.B., Fontes W., Sousa M.V., Filho E.X.F., 2000 Purification and characterization of a new xylanase from Acrophialophora nainiana Journal of Biotechnology 81:199–204
Sá-Pereira P., Paveia H., Costa Ferreira M., Aires-Barros M.R., 2003 A new look at xylanases Molecular Biotechnology 24:257–281
Silveira F.Q.P., Melo I.S., Filho E.X.F., 1997 Carbohydrate-hydrolysing enzyme activity producing by solid-state cultures of Trichoderma harzianum strains Brazilian Journal of Microbiology 28:152–156
Tosi L.R., Terenzi H.F., Jorge J.A., 1993 Purification and characterization of an extracellular glucoamylase from the thermophilic fungus Humicola grisea var. thermoidea Canadian Journal of Microbiology 39:846–852
Ximenes F.A., Silveira F.Q.P., Filho E.X.F., 1996 Production of β-xylosidase activity by Trichoderma harzianum strains Current Microbiology 33:71–77
Ximenes F.A., Fonseca A.S., Ximenes E.A., Silveira F.Q.P., Silva C.H.C., Lucena-Neto S.A., Ribeiro W.R.C., Filho E.X.F., 1997 Xylan-degrading enzyme production by solid-state cultures of aerobic fungi Brazilian Journal of Microbiology 28:22–28
Zanoelo F.F., Polizeli M.L.T.M., Terenzi H.F., Jorge J.A., 2004 Purification and biochemical properties of a thermostable xylose-tolerant β-D-xylosidase from Scytalidum thermophilum Journal of Industrial Microbiology and Biotechnology 31:170–176
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E.X.F.F. and T. I. acknowledge the receipt of research fellowship and post-graduate maintenance scholarship from CNPq (Brazil) and CAPES (Brazil), respectively.
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Iembo, T., Azevedo, M., Jr., C.B. et al. Purification and partial characterization οf a new β-xylosidase from Humicola grisea var. thermoidea . World J Microbiol Biotechnol 22, 475–479 (2006). https://doi.org/10.1007/s11274-005-9059-3
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DOI: https://doi.org/10.1007/s11274-005-9059-3