Skip to main content
SpringerLink
Log in

Purification and characterization of a novel β-1,3/1,4-glucanase from Sistotrema brinkmannii HQ717718

  • Original Article
  • Published:
Journal of the Korean Society for Applied Biological Chemistry Submit manuscript

Abstract

A highly efficient extracellular β-1,3/1,4-glucanase was purified from the culture broth of Sistotrema brinkmannii HQ717718. The molecular mass of β-1,3/1,4-glucanase was respectively determined to be 83 and 166 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and gel filtration chromatography, indicating that the enzyme is a dimer. The optimum activity of β-1,3/1,4-glucanase against Avicel was observed at pH 4.0 and 65°C. Under the same conditions, V max, K m, and k cat values for Avicel were 136.5 U · mg−1 of protein, 3.8 mM, and 211 s−1, respectively. Furthermore, the DNA sequence of gene coding the enzyme showed a significant homology with hydrolases from the glycoside hydrolase family 55. Although β-1,3/1,4-glucanases have been purified and characterized from several other sources, S. brinkmannii β-1,3/1,4-glucanase is distinct from other β-1,3/1,4-glucanases due to its high catalytic efficiency toward Avicel and broad substrate specificity.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Bradford MM (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.

    Article  CAS  Google Scholar 

  • Carpita NC (1996). Structure and biogenesis of the cell walls of grasses. Annu Rev Plant Physiol Plant Mol Biol 47, 445–476.

    Article  CAS  Google Scholar 

  • Celestino K, Cunha R, and Felix C (2006). Characterization of a β-glucanase produced by Rhizopus microsporus var. microsporus, and its potential for application in the brewing industry. BMC Biochem 7, 1–9.

    Article  Google Scholar 

  • Cheng YM, Hong TY, Liu CC, and Meng MS (2009). Cloning and functional characterization of a complex endo-β-1,3-glucanase from Paenibacillus sp. Appl Microbiol Biot 81, 1051–1061.

    Article  CAS  Google Scholar 

  • Erfle JD, Teather RM, Wood PJ, and Irvin JE (1988). Purification and Properties of a 1,3–1,4-β-d-glucanase (lichenase, 1,3–1,4-β-d-glucan 4-glucanohydrolase, EC-3.2.1.73) from Bacteroides succinogenes cloned in Escherichia coli. Biochem J 255, 833–841.

    CAS  Google Scholar 

  • Fuchs KP, Zverlov VV, Velikodvorskaya GA, Lottspeich F, and Schwarz WH (2003). Lic16A of Clostridium thermocellum, a non-cellulosomal, highly complex endo-β-1,3-glucanase bound to the outer cell surface. Microbiol-Sgm 149, 1021–1031.

    Article  CAS  Google Scholar 

  • Genta FA, Bragatto I, Terra WR, and Ferreira C (2009). Purification, characterization and sequencing of the major β-1,3-glucanase from the midgut of Tenebrio molitor larvae. Insect Biochem Molec 39, 861–874.

    Article  CAS  Google Scholar 

  • Gorlach JM, Van der Knaap E, and Walton JD (1998). Cloning and targeted disruption of MLG1, a gene encoding two of three extracellular mixedlinked glucanases of Cochliobolus carbonum. Appl Environ Microbiol 64, 385–391.

    CAS  Google Scholar 

  • Gosalbes MJ, Perezgonzalez JA, Gonzalez R, and Navarro A (1991). 2 β-Glycanase genes are clustered in Bacillus polymyxa — Molecular cloning, expression, and sequence analysis of genes encoding a xylanase and an endo-β-(1,3)-(1,4)-glucanase. J Bacteriol 173, 7705–7710.

    CAS  Google Scholar 

  • Grishutin SG, Gusakov AV, Dzedzyulya EI, and Sinitsyn AP (2006). A lichenase-like family 12 endo-(l→4)-β-glucanase from Aspergillus japonicus: study of the substrate specificity and mode of action on betaglucans in comparison with other glycoside hydrolases. Carbohyd Res 341, 218–229.

    Article  CAS  Google Scholar 

  • Hamada N, Ishikawa K, Fuse N, Kodaira R, Shimosaka M, Amano Y et al. (1999). Purification, characterization and gene analysis of exo-cellulase II (Ex-2) from the white rot basidiomycete Irpex lacteus. J Biosci Bioeng 87, 442–451.

    Article  CAS  Google Scholar 

  • Jeya M, Joo AR, Lee KM, Tiwari MK, Lee KM, Kim SH et al. (2010). Characterization of β-glucosidase from a strain of Penicillium purpurogenum KJS506. Appl Microbiol Biot 86, 1473–1484.

    Article  CAS  Google Scholar 

  • Kanokratana P, Chantasingh D, Champreda V, Tanapongpipat S, Pootanakit K, and Eurwilaichitr L (2008). Identification and expression of cellobiohydrolase (CBHI) gene from an endophytic fungus, Fusicoccum sp (BCC4124) in Pichia pastoris. Protein Expres Purif 58, 148–153.

    Article  CAS  Google Scholar 

  • Kaur J, Chadha BS, Kumar BA, and Saini HS (2007). Purification and characterization of two endoglucanases from Melanocarpus sp MTCC 3922. Bioresource Technol 98, 74–81.

    Article  CAS  Google Scholar 

  • Kawai R, Igarashi K, and Samejima M (2006). Gene cloning and heterologous expression of glycoside hydrolase family 55 β-1,3-glucanase from the basidiomycete Phanerochaete chrysosporium. Biotechnol Lett 28, 365–371.

    Article  CAS  Google Scholar 

  • Laemmli UK and others (1970). Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227, 680–685.

    Article  CAS  Google Scholar 

  • Lee KM, Joo AR, Jeya M, Lee KM, Moon HJ, and Lee JK (2011a). Production and characterization of cellobiohydrolase from a novel strain of Penicillium purpurogenum KJS506. Appl Biochem Biotech 163, 25–39.

    Article  CAS  Google Scholar 

  • Lee KM, Moon HJ, Kalyani D, Kim H, Kim IW, Jeya M et al. (2011b). Characterization of cellobiohydrolase from a newly isolated strain of Agaricus arvencis. J Microbiol Biotechnol 21, 711–718.

    Article  Google Scholar 

  • Lin J, Pillay B, and Singh S (1999). Purification and biochemical characteristics of β-d-glucosidase from a thermophilic fungus, Thermomyces lanuginosus-SSBP. Biotechnol Appl Bioc 30, 81–87.

    CAS  Google Scholar 

  • Loprete DM and Hill TW (2002). Isolation and characterization of an endo-(1,4)-β-glucanase secreted by Achlya ambisexualis. Mycologia 94, 903–911.

    Article  CAS  Google Scholar 

  • Louw ME, Reid SJ, and Watson TG (1993). Characterization, cloning and sequencing of a thermostable endo-(1,3-1,4) β-glucanase-encoding gene from an alkalophilic Bacillus brevis. Appl Microbiol Biotechnol 38, 507–513.

    Article  CAS  Google Scholar 

  • McCarthy T, Hanniffy O, Savage AV, and Tuohy MG (2003). Catalytic properties and mode of action of three endo-β-glucanases from Talaromyces emersonii on soluble β-1,4- and β-1,3;1,4-linked glucans. Int J Biol Macromol 33, 141–148.

    Article  CAS  Google Scholar 

  • Miller GL (1959). Use of dinitrosalicylic acid reagent for determination of reducing sugar. Anal Chem 31, 426–428.

    Article  CAS  Google Scholar 

  • Planas N (2000). Bacterial 1,3–1,4-β-glucanases: structure, function and protein engineering. Bba-Protein Struct M 1543, 361–382.

    Article  CAS  Google Scholar 

  • Saitou N and Nei M (1987). The neighbor-joining method — a new method for reconstructing phylogenetic trees. Mol Biol Evol 4, 406–425.

    CAS  Google Scholar 

  • Sakamoto Y, Nakade K, and Sato T (2009). Characterization of the post-harvest changes in gene transcription in the gill of the Lentinula edodes fruiting body. Curr Genet 55, 409–423.

    Article  CAS  Google Scholar 

  • Shin K, Kim YH, Jeya M, Lee JK, and Kim YS (2010). Purification and characterization of a thermostable cellobiohydrolase from Fomitopsis pinicola. J Microbiol Biotechn 20, 1681–1688.

    CAS  Google Scholar 

  • Shoemaker S, Schweickart V, Ladner M, Gelfand D, Kwok S, Myambo K et al. (1983). Molecular cloning of exo-cellobiohydrolase-I derived from Trichoderma reesei strain-L27. Bio-Technol 1, 691–696.

    Article  CAS  Google Scholar 

  • Tamura K, Dudley J, Nei M, and Kumar S (2007). MEGA4: Molecular evolutionary genetics analysis (MEGA) software version 4.0. Mol Biol Evol 24, 1596–1599.

    Article  CAS  Google Scholar 

  • Teng D, Fan Y, Yang YL, Tian ZG, Luo J, and Wang JH (2007). Codon optimization of Bacillus licheniformis β-1,3–1,4-glucanase gene and its expression in Pichia pastoris. Appl Microbiol Biot 74, 1074–1083.

    Article  CAS  Google Scholar 

  • Tuohy MG, Walsh DJ, Murray PG, Claeyssens M, Cuffe MM, Savage AV et al. (2002). Kinetic parameters and mode of action of the cellobiohydrolases produced by Talaromyces emersonii. Bba-Protein Struct M 1596, 366–380.

    Article  CAS  Google Scholar 

  • Yan TR and Lin CL (1997). Purification and characterization of a glucosetolerant β-glucosidase from Aspergillus niger CCRC 31494. Biosci Biotechnol Biochem 61, 965–970.

    Article  CAS  Google Scholar 

  • Yanbin T, Shaoqing Y, Qiaojuan Y, Peng Z, Jian C, and Zhengqiang J (2012). Purification and characterization of a novel β-1,3–1,4-glucanase (Lichenase) from thermophilic Rhizomucor miehei with high specific activity and its gene sequence. J Agr Food Chem 60(9), 2354–2361.

    Article  Google Scholar 

  • Yang SQ, Yan QJ, Jiang ZQ, Fan GS, and Wang L (2008). Biochemical characterization of a novel thermostable β-1,3–1,4-glucanase (Lichenase) from Paecilomyces thermophila. J Agr Food Chem 56, 5345–5351.

    Article  CAS  Google Scholar 

  • Ye XY, Ng TB, and Cheng KJ (2001). Purification and characterization of a cellulase from the ruminal fungus Orpinomyces joyonii cloned in Escherichia coli. Int J Biochem Cell B 33, 87–94.

    Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Chemical Engineering, Konkuk University, Seoul, 143-701, Republic of Korea

    Zongpei Zhao, Jung-Kul Lee & In-Won Kim

  2. Institute of SK-KU Biomaterials, Konkuk University, Seoul, 143-701, Republic of Korea

    Priyadharshini Ramachandran, Jung-Kul Lee & In-Won Kim

  3. Department of Food Science and Biotechnology, Wonkwang University, Iksan-city, JeonBuk, 570-749, Republic of Korea

    Joon-Ho Choi

Authors
  1. Zongpei Zhao
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Priyadharshini Ramachandran
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Joon-Ho Choi
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Jung-Kul Lee
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. In-Won Kim
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding authors

Correspondence to Jung-Kul Lee or In-Won Kim.

Electronic supplementary material

Rights and permissions

Reprints and permissions

About this article

Cite this article

Zhao, Z., Ramachandran, P., Choi, JH. et al. Purification and characterization of a novel β-1,3/1,4-glucanase from Sistotrema brinkmannii HQ717718. J Korean Soc Appl Biol Chem 56, 263–270 (2013). https://doi.org/10.1007/s13765-013-3028-6

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s13765-013-3028-6

Keywords

Search

Navigation