Skip to main content
SpringerLink
Log in

Characterization of endo-β-mannanase from Enterobacter ludwigii MY271 and application in pulp industry

  • Original Paper
  • Published:
Bioprocess and Biosystems Engineering Aims and scope Submit manuscript

Abstract

β-Mannanases are the second most important enzymes for the hydrolysis of hemicelluloses. An endo-β-mannanase from Enterobacter ludwigii MY271 was purified at 11.7 ± 0.2-fold to homogeneity with a final recovery of 15.2 ± 0.2 %. Using purified β-mannanase protein and SDS-PAGE, the molecular mass was found to be 43.16 kDa. The optimal pH and temperature of the enzyme was found to be 7.0 and 55 °C, respectively. The β-mannanase activity was stable over a broad pH range of pH 2.0–10.0. In addition, the purified enzyme was highly activated by several metal ions and chemical reagents, such as Mg2+, l-cysteine, glutathione (GSH) and β-mercaptoethanol. Whereas the enzyme was strongly inhibited by Hg2+, Cu2+, N-bromosuccinimide (NBS), 1-ethyl-3-(3-dimethyl-amino-propyl)-carbodiimide (EDC), phenylmethanesulfonyl fluoride (PMSF), and sodium dodecyl sulfate (SDS). The β-mannanase was highly active towards glucomannan, and showed endo-activity by producing a mixture of oligosaccharides. Moreover, the enzyme displayed a classical endo-type mode on mannooligosaccharides. The β-mannanase coupled with xylanase significantly improved the brightness of kraft pulp, whereas it has no remarkable effect on the tensile strength of the pulp. Our functional studies of the purified β-mannanase indicate that the enzyme is beneficial to industrial applications, in particular, biotechnological processes, such as food, feed and pulp industry.

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

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

References

  1. Dhawan S, Kaur J (2007) Microbial mannanases: an overview of production and applications. Crit Rev Biotechnol 27:197–216

    Article  CAS  Google Scholar 

  2. Petkowicz CLO, Reicher F, Chanzy H, Taravel FR, Vuong R (2001) Linear mannan in the endosperm of Schizolobium amazonicum. Carbohydr Polym 44:107–112

    Article  CAS  Google Scholar 

  3. Chauhan PS, Sharma P, Puri N, Gupta N (2014) A process for reduction in viscosity of coffee extract by enzymatic hydrolysis of mannan. Bioprocess Biosyst Eng 37:1459–1467

    Article  CAS  Google Scholar 

  4. Hägglund P, Eriksson T, Collén A, Nerinckx W, Claeyssens M, Stålbrand H (2003) A cellulose-binding module of the Trichoderma reesei β-mannanase Man5A increases the mannan-hydrolysis of complex substrates. J Biotechnol 101:37–48

    Article  Google Scholar 

  5. Moreira LRS (2008) An overview of mannan structure and mannan-degrading enzyme systems. Appl Microbiol Biotechnol 79:165–178

    Article  CAS  Google Scholar 

  6. Van Laere KMJ, Hartemink R, Beldman G, Pitson S, Dijkema C, Schols HA, Voragen AGJ (1999) Transglycosidase activity of Bifidobacterium adolescentis DSM 20083 α-galactosidase. Appl Microbiol Biotechnol 52:681–688

    Article  Google Scholar 

  7. Clarke JH, Davidson K, Rixon JE, Halstead JR, Fransen MP, Gilbert HJ, Hazlewood GP (2000) A comparison of enzyme-aided bleaching of softwood paper pulp using combinations of xylanase, mannanase and α-galactosidase. Appl Microbiol Biotechnol 53:661–667

    Article  CAS  Google Scholar 

  8. Kansoh AL, Nagieb ZA (2004) Xylanase and mannanase enzymes from Streptomyces galbus NR and their use in biobleaching of softwood kraft pulp. Antonie Van Leeuwenhoek 85:103–114

    Article  CAS  Google Scholar 

  9. Fan Z, Wagschal K, Chen W, Montross MD, Lee CC, Yuan L (2009) Multimeric hemicellulases facilitate biomass conversion. Appl Environ Microbiol 75:1754–1757

    Article  CAS  Google Scholar 

  10. Chen J, Liu D, Shi B, Wang H, Cheng Y, Zhang W (2009) Optimization of hydrolysis conditions for the production of glucomanno-oligosaccharides from konjac using β-mannanase by response surface methodology. Carbohydr Polym 93:81–88

    Article  Google Scholar 

  11. Luo L, Cai J, Wang C, Lin J, Du X, Zhou A, Xiang M (2015) Purification and characterization of an alkaliphilic endo-xylanase from Streptomyces althioticus LMZM and utilization in the pulp paper industry. J Chem Technol Biotechnol. doi:10.1002/jctb.4690

    Google Scholar 

  12. Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685

    Article  CAS  Google Scholar 

  13. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193:265–275

    CAS  Google Scholar 

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

    Article  CAS  Google Scholar 

  15. Lineweaver H, Burk D (1934) The determination of enzyme dissociation constants. J Am Chem Soc 56:658–666

    Article  CAS  Google Scholar 

  16. Zang H, Xie S, Wu H, Wang W, Shao X, Wu L, Gao X (2015) A novel thermostable GH5_7 β-mannanase from Bacillus pumilus GBSW19 and its application in manno-oligosaccharides (MOS) production. Enzym Microb Technol 78:1–9

    Article  CAS  Google Scholar 

  17. Liu Q, Yang P, Luo H, Shi P, Huang H, Meng K, Yao B (2012) A novel endo-1, 4-β-mannanase from Bispora antennata with good adaptation and stability over a broad pH range. Appl Biochem Biotechnol 166:1442–1453

    Article  CAS  Google Scholar 

  18. Lee YS, Zhou Y, Park IH, Chandra MRGS, Ahn SC, Choi YL (2010) Isolation and purification of thermostable β-mannanase from Paenibacillus illinoisensis ZY-08. J Korean Soc Appl Biol Chem 53:1–7

    Article  CAS  Google Scholar 

  19. Zhang M, Chen XL, Zhang ZH, Sun CY, Chen LL, He HL, Zhang YZ (2009) Purification and functional characterization of endo-β-mannanase MAN5 and its application in oligosaccharide production from konjac flour. Appl Microbiol Biotechnol 83:865–873

    Article  CAS  Google Scholar 

  20. Chauhan PS, Sharma P, Puri N, Gupta N (2014) Purification and characterization of an alkali-thermostable β-mannanase from Bacillus nealsonii PN-11 and its application in mannooligosaccharides preparation having prebiotic potential. Eur Food Res Technol 238:927–936

    Article  CAS  Google Scholar 

  21. Blibech M, Ghorbel RE, Fakhfakh I, Ntarima P, Piens K, Bacha AB, Chaabouni SE (2010) Purification and characterization of a low molecular weight of β-mannanase from Penicillium occitanis Pol6. Appl Biochem Biotechnol 160:1227–1240

    Article  CAS  Google Scholar 

  22. Khandeparkar R, Bhosle NB (2006) Purification and characterization of thermoalkalophilic xylanase isolated from the Enterobacter sp. MTCC 5112. Res Microbiol 157:315–325

    Article  CAS  Google Scholar 

  23. Yang H, Shi P, Lu H, Wang H, Luo H, Huang H, Yao B (2015) A thermophilic β-mannanase from Neosartorya fischeri P1 with broad pH stability and significant hydrolysis ability of various mannan polymers. Food Chem 173:283–289

    Article  CAS  Google Scholar 

  24. Ferreira HM, Filho EXF (2004) Purification and characterization of a β-mannanase from Trichoderma harzianum strain T4. Carbohydr Polym 57:23–29

    Article  CAS  Google Scholar 

  25. Yin LJ, Tai HM, Jiang ST (2012) Characterization of mannanase from a novel mannanase-producing bacterium. J Agric Food Chem 60:6425–6431

    Article  CAS  Google Scholar 

  26. Naganagouda K, Salimath PV, Mulimani VH (2009) Purification and characterization of endo-beta-1, 4 mannanase from Aspergillus niger gr for application in food processing industry. J Microbiol Biotechnol 19:1184–1190

    CAS  Google Scholar 

  27. Jiang Z, Wei Y, Li D, Li L, Chai P, Kusakabe I (2006) High-level production, purification and characterization of a thermostable β-mannanase from the newly isolated Bacillus subtilis WY34. Carbohydr Polym 66:88–96

    Article  CAS  Google Scholar 

  28. Chauhan PS, Puri N, Sharma P, Gupta N (2012) Mannanases: microbial sources, production, properties and potential biotechnological applications. Appl Microbiol Biotechnol 93:1817–1830

    Article  CAS  Google Scholar 

  29. Chauhan PS, Soni SK, Sharma P, Saini A, Gupta N (2014) A mannanase from Bacillus nealsonii PN-11: statistical optimization of production and application in biobleaching of pulp in combination with xylanase. Int J Pharma Biosci 5:237–251

    CAS  Google Scholar 

  30. Mabrouk MEM, El Ahwany AMD (2008) Production of β-mannanase by Bacillus amylolequifaciens 10A1 cultured on potato peels. Afr J Biotechnol 7:1123–1128

    CAS  Google Scholar 

  31. Meenakshi M, Singh G, Bhalla A, Hoondal G (2010) Solid state fermentation and characterization of partially purified thermostable mannanase from Bacillus sp. MG-33. Bioresource 5:1689–1701

    CAS  Google Scholar 

  32. Kote NV, Patil AGG, Mulimani VH (2009) Optimization of the production of thermostable endo-β-1, 4 mannanases from a newly isolated Aspergillus niger gr and Aspergillus flavus gr. Appl Biochem Biotechnol 152:213–223

    Article  CAS  Google Scholar 

  33. Wang C, Luo H, Niu C, Shi P, Huang H, Meng K, Yao B (2015) Biochemical characterization of a thermophilic β-mannanase from Talaromyces leycettanus JCM12802 with high specific activity. Appl Microbiol Biotechnol 99:1217–1228

    Article  CAS  Google Scholar 

Download references

Acknowledgments

This research was supported by the Key Laboratory of Fermentation engineering (Ministry education), Hubei University of Technology, China.

Author information

Authors and Affiliations

  1. Key Laboratory of Fermentation Engineering (Ministry of Education), Hubei Provincial Cooperative Innovation Center of Industrial Fermentation, Hubei University of Technology, Wuhan, 430068, China

    Miao Yang, Jun Cai, Changgao Wang, Xin Du & Jianguo Lin

Authors
  1. Miao Yang
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Jun Cai
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Changgao Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Xin Du
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Jianguo Lin
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Jun Cai.

Ethics declarations

Conflict of interest

The authors declare that there is no conflict of interests regarding the publication of this paper.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Yang, M., Cai, J., Wang, C. et al. Characterization of endo-β-mannanase from Enterobacter ludwigii MY271 and application in pulp industry. Bioprocess Biosyst Eng 40, 35–43 (2017). https://doi.org/10.1007/s00449-016-1672-z

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00449-016-1672-z

Keywords

Search

Navigation