Applied Microbiology and Biotechnology

, Volume 86, Issue 5, pp 1473–1484 | Cite as

Characterization of β-glucosidase from a strain of Penicillium purpurogenum KJS506

  • Marimuthu Jeya
  • Ah-Reum Joo
  • Kyoung-Mi Lee
  • Manish Kumar Tiwari
  • Kyoung-Min Lee
  • Sang-Hwan Kim
  • Jung-Kul Lee
Biotechnologically Relevant Enzymes and Proteins


A novel β-glucosidase (BGL)-producing strain was isolated and identified as Penicillium purpurogenum KJS506 based on its morphology and internal transcribed spacer (ITS) rDNA gene sequence. When rice straw and corn steep powder were used as carbon and nitrogen sources, respectively, the maximal BGL activity of 12.3 U ml−1, one of the highest levels among BGL-producing microorganisms was observed. The optimum temperature and pH for BGL production were 32 °C and 4, respectively. An extracellular BGL was purified to homogeneity by sequential chromatography of P. purpurogenum culture supernatants, and the purified BGL showed higher activity (V max = 934 U mg protein–1) than most BGLs from other sources. The complete ORF of bgl3 was cloned from P. purpurogenum by a modified thermal asymmetric interlaced polymerase chain reaction. The bgl3 gene consists of a 2,571-bp ORF and encodes a putative protein containing 856 amino acids with a calculated molecular mass of 89,624 Da. The putative gene product was identified as a member of glycoside hydrolase family 3. The present results should contribute to improved industrial production of BGL by P. purpurogenum KJS506.


Cloning β-Glucosidase Homology modeling Penicillium purpurogenum TAIL-PCR 



This study was supported by a grant (code 2008A0080126) from ARPC. This subject was also supported by Korea Ministry of Environment as “The GAIA Project”.


  1. Altschul SF, Gish W, Miller W, Myers EW, Lipman DJ (1990) Basic local alignment search tool. J Mol Biol 215:403–410Google Scholar
  2. Arja MJL, Vesa J, Raija L (2004) Three cellulases from Melanocarpus albomyces for textile treatment at neutral pH. Enzyme Microb Technol 34:332–341CrossRefGoogle Scholar
  3. Bahia A, Ali G (2006) Characterization of novel beta-glucosidase from a stachybotrys strain. Appl Microbiol Biotechnol 32:191–197Google Scholar
  4. Beguin P, Aubert JP (1994) The biological degradation of cellulose. FEMS Microbiol Rev 13:25–58CrossRefGoogle Scholar
  5. Bhat MK, Bhat S (1997) Cellulose degrading enzymes and their potential industrial applications. Biotechnol Adv 15:583–620CrossRefGoogle Scholar
  6. Coulibaly L, Naveau H, Agathos SN (2002) A tanks-in-series bioreactor to simulate macromolecule-laden wastewater pretreatment under sewer conditions by Aspergillus niger. Water Res 36:3941–3948CrossRefGoogle Scholar
  7. Daenen L, Saison D, Sterckx F, Delvaux FR, Verachtert H, Derdelinckx G (2008) Screening and evaluation of the glucoside hydrolase activity in Saccharomyces and Brettanomyces brewing yeasts. J Appl Microbiol 104:478–88Google Scholar
  8. Da-Silva R, Gomes E, Franco CML (1997) Pectinases, hemicelulase e cellulases substrate, production application no processamento de alimentos. Bol SBCTA 31:249–250Google Scholar
  9. de Palma-Fernandez ER, Gomes E, da Silva R (2002) Purification and characterization of two beta-glucosidases from the thermophilic fungus Thermoascus aurantiacus. Folia Microbiol (Praha) 47:685–690CrossRefGoogle Scholar
  10. Eriksson KE, Pettersson B, Westermark U (1974) Oxidation: an important enzyme reaction in fungal degradation of cellulose. FEBS Lett 49:282–285CrossRefGoogle Scholar
  11. Galas E, Romanowska I (1996) Production purification and characterization of a highly glucose-tolerant novel ß-glucosidase from Candida peltata. Appl Environ Microbiol 62:3165–3170Google Scholar
  12. Gasteiger E, Gattiker A, Hoogland C, Ivanyi I, Appel RD, Bairoch A (2003) ExPASy: The proteomics server for in-depth protein knowledge and analysis. Nucleic Acids Res 31:3784–3788CrossRefGoogle Scholar
  13. Henrissat B (1998) Glycosidase families. Biochem Soc Trans 26:153–156Google Scholar
  14. Henrissat B, Callebaut I, Fabrega S, Lehn P, Mornon JP, Davies G (1995) Conserved catalytic machinery and the prediction of a common fold for several families of glycosyl hydrolases. Proc Natl Acad Sci U.S.A. 92:7090–7094CrossRefGoogle Scholar
  15. Himmel ME, Adney WS, Fox JW, Mitchell DJ, Baker JO (1993) Isolation and characterization of two forms of beta-D-glucosidase from Aspergillus niger. Appl Biochem Biotechnol 39–40:213–225CrossRefGoogle Scholar
  16. Joo AR et al (2009) Purification and characterization of a beta-1,4-glucosidase from a newly isolated strain of Fomitopsis pinicola. Appl Microbiol Biotechnol 83:285–294CrossRefGoogle Scholar
  17. Karnchanatat A et al (2007) Purification and biochemical characterization of an extracellular beta-glucosidase from the wood-decaying fungus Daldinia eschscholzii (Ehrenb.:Fr.) Rehm. FEMS Microbiol Lett 270:162–170CrossRefGoogle Scholar
  18. Kaur J, Bhupinder SC, Badhan AK, Ghatora KS (2007) Purification and characterization of β-glucosidase from Melanocarpus sp MTCC 3922. Electron J Biotechnol 10:261–270CrossRefGoogle Scholar
  19. Kempton JB, Withers SG (1992) Mechanism of Agrobacterium beta-glucosidase: kinetic studies. Biochemistry 31:9961–9969CrossRefGoogle Scholar
  20. Kuriyama K, Tsuchiya K, Murui T (1995) Some properties of transglycosylation activity of sesame β-glucosidase. Biosci Biotechnol Biochem 59:1142–1143CrossRefGoogle Scholar
  21. Leclerc MAA, Ratomahenina R, Galzy P (1987) Yeast β-glucosidases. Biotechnol Genet Eng Rev 5:269–295Google Scholar
  22. Leite RSR, Gomes E, Da-Silva R (2007) Characterization and comparison of thermostability of purified β-glucosidases from a mesophilic Aureobasidium pullulans and thermophilic Thermoascus aurantiacus. Process Biochem 42:1101–1106CrossRefGoogle Scholar
  23. Lin J, Balakrishna P, Suren S (1999) Purification and biochemical characterization of β-glucosidase from a thermophilic fungus, Thermomyces lanuginosus - SSBP. Biotechnol Appl Biochem 30:81–87Google Scholar
  24. Liu YG, Huang N (1998) Efficient amplification of insert end sequences from bacterial artificial chromosome clones by thermal asymmetric interlaced PCR. Plant Mol Bio Rep 16:175–181CrossRefGoogle Scholar
  25. Liu YG, Whittier RF (1995) Thermal asymmetric interlaced PCR: Automatable amplification and sequencing of insert end fragments from P1 and YAC clones for chromosome walking. Genomics 25:674–681CrossRefGoogle Scholar
  26. Lymar ES, Li B, Renganathan V (1995) Purification and characterization of a cellulose-binding (beta)-glucosidase from cellulose-degrading cultures of Phanerochaete chrysosporium. Appl Environ Microbiol 61:2976–2980Google Scholar
  27. Mach R, Zeilinger S (2003) Regulation of gene expression in industrial fungi Trichoderma. Appl Microbiol biotechnol 60:515–522Google Scholar
  28. Magalhaes PO, Ferraz A, Milagres AF (2006) Enzymatic properties of two β-glucosidases from Ceriporiopsis subvermispora produced in biopulping conditions. J Appl Microbiol 101:480–486CrossRefGoogle Scholar
  29. Makoto M, Isao O, Sakuzo F, Ichiro Y (1988) Nucleotide sequences of Saccharomycopsis fibuligera genes for extracellular β-glucosidases as expressed in Saccharomyces cerevisiae. Appl Microbiol Biotechnol 54:3147–3155Google Scholar
  30. Rashid MH, Siddiqui KS (1997) Purification and characterization of a beta-glucosidase from Aspergillus niger. Folia Microbiol (Praha) 42:544–550CrossRefGoogle Scholar
  31. Saha BC, Freer SN, Bothast RJ (1994) Production, purification, and properties of a thermostable beta-glucosidase from a color variant strain of Aureobasidium pullulans. Appl Environ Microbiol 60:3774–3780Google Scholar
  32. Saloheimo M, Nakari-Setala T, Tenkanen M, Penttila M (1997) cDNA cloning of a Trichoderma reesei cellulase and demonstration of endoglucanase activity by expression in yeast. Eur J Biochem 249:584–591CrossRefGoogle Scholar
  33. Shevchenko A, Wilm M, Vorm O, Mann M (1996) Mass spectrometric sequencing of proteins from silver-stained polyacrylamide gels. Anal Chem 68:850–858CrossRefGoogle Scholar
  34. Shinoyama HTV, Ando A, Fujii T, Sasaki M, Doi Y, Yasui T (1991) Enzymatic synthesis of useful alkyl-β-glucosides. Agri Biol Chem 55:1679–1681Google Scholar
  35. Stahl P, Klug M (1996) Characterization and differentiation of filamentous fungi based on fatty acid composition. Appl Environ Microbiol 62:4136–4146Google Scholar
  36. Valaskova V, Baldrian P (2006) Degradation of cellulose and hemicelluloses by the brown rot fungus Piptoporus betulinus-production of extracellular enzymes and characterization of the major cellulases. Microbiology 152:3613–3622CrossRefGoogle Scholar
  37. Wei DL, Kirimura K, Usami S, Lin TH (1996) Purification and characterization of an extracellular beta-glucosidase from the wood-grown fungus Xylaria regalis. Curr Microbiol 33:297–301CrossRefGoogle Scholar
  38. Wong WK, Ali A, Chan WK, Ho V, Lee NT (1998) The cloning, expression and characterization of a cellobiase gene encoding a secretory enzyme from Cellulomonas biazotea. Gene 207:79–86CrossRefGoogle Scholar
  39. Workman WE, Day DF (1982) Purification and properties of beta-glucosidase from Aspergillus terreus. Appl Environ Microbiol 44:1289–1295Google Scholar
  40. Yan TS, Lin CL (1996) Purification and characterization of a glucose-tolerant β-glucosidase from Aspergillus niger CCRC31494. Biosci Biotechnol Biochem 61:965–970CrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2009

Authors and Affiliations

  • Marimuthu Jeya
    • 1
  • Ah-Reum Joo
    • 2
  • Kyoung-Mi Lee
    • 1
    • 3
  • Manish Kumar Tiwari
    • 1
  • Kyoung-Min Lee
    • 1
  • Sang-Hwan Kim
    • 1
  • Jung-Kul Lee
    • 1
    • 3
  1. 1.Department of Chemical EngineeringKonkuk UniversitySeoulSouth Korea
  2. 2.Department of Bioscience and BiotechnologyKonkuk UniversitySeoulSouth Korea
  3. 3.Institute of Biomedical Science and TechnologyKonkuk UniversitySeoulSouth Korea

Personalised recommendations