Skip to main content
SpringerLink
Log in

Biochemical characterization of thermostable β-1,4-mannanase belonging to the glycoside hydrolase family 134 from Aspergillus oryzae

  • Biotechnologically relevant enzymes and proteins
  • Published:
Applied Microbiology and Biotechnology Aims and scope Submit manuscript

Abstract

A β-1,4-mannanase, termed AoMan134A, that belongs to the GH 134 family was identified in the filamentous fungus Aspergillus oryzae. Recombinant AoMan134A was expressed in Pichia pastoris, and the purified enzyme produced mannobiose, mannotriose, mannotetraose, and mannopentaose from galactose-free β-mannan, with mannotriose being the predominant reaction product. The catalytic efficiency (k cat/K m ) of AoMan134A was 6.8-fold higher toward galactomannan from locust bean gum, than toward galactomannan from guar gum, but similar toward galactomannan from locust bean gum and glucomannan from konjac flour. After incubation at 70°C for 120 min, the activity of AoMan134A toward glucomannan decreased to 50% of the maximal activity at 30°C. AoMan134A retained 50% of its β-1,4-mannanase activity after heating at 90°C for 30 min, indicating that AoMan134A is thermostable. Furthermore, AoMan134A was stable within a neutral-to-alkaline pH range, as well as exhibiting stability in the presence of a range of organic solvents, detergents, and metal ions. These findings suggest that AoMan134A could be useful in a diverse range of industries where conversion of β-mannans is of prime importance.

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
Fig. 5

Similar content being viewed by others

References

  • Aspinall GO (1959) Structural chemistry of the hemicelluloses. Adv Carbohydr Chem 14:429–468

    CAS  PubMed  Google Scholar 

  • Barak S, Mudgil D (2014) Locust bean gum: processing, properties and food applications: a review. Int J Biol Macromol 66:74–80

    Article  CAS  PubMed  Google Scholar 

  • Bien-Cuong D, Thi-Thu D, Berrin JG, Haltrich D, Kim-Anh T, Sigoillot JC, Yamabhai M (2009) Cloning, expression in Pichia pastoris, and characterization of a thermostable GH5 mannan endo-1,4-β-mannosidase from Aspergillus niger BK01. Microb Cell Factories 8:59

    Article  Google Scholar 

  • Cantarel BL, Coutinho PM, Rancurel C, Bernard T, Lombard V, Henrissat B (2009) The carbohydrate-active EnZymes database (CAZy): an expert resource for glycogenomics. Nucleic Acids Res 37:233–238

    Article  Google Scholar 

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

    Article  CAS  PubMed  Google Scholar 

  • 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 

  • Chiyanzu I, Brienzo M, García-Aparicio MP, Görgens JF (2014) Application of endo-β-1,4-D-mannanase and cellulase for the release of mannooligosaccharides from steam-pretreated spent coffee ground. Appl Biochem Biotechnol 172:3538–3557

    Article  CAS  PubMed  Google Scholar 

  • Comfort DA, Chhabra SR, Conners SB, Chou CJ, Epting KL, Johnson MR, Jones KL, Sehgal AC, Kelly RM (2004) Strategic biocatalysis with hyperthermophilic enzymes. Green Chem 6:459–465

    Article  CAS  Google Scholar 

  • de Vries RP, Visser J (2001) Aspergillus enzymes involved in degradation of plant cell wall polysaccharides. Microbiol Mol Biol Rev 65:497–522

    Article  PubMed  PubMed Central  Google Scholar 

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

    Article  CAS  PubMed  Google Scholar 

  • Duffaud GD, McCutchen CM, Leduc P, Parker KL, Kelly RM (1997) Purification and characterization of extremely thermostable β-mannanase, β-mannosidase, and α-galactosidase from the hyperthermophilic eubacterium Thermotoga neapolitana 5068. Appl Environ Microbiol 63:169–177

    CAS  PubMed  PubMed Central  Google Scholar 

  • Duruksu G, Ozturk B, Biely P, Bakir U, Ogel ZB (2009) Cloning, expression and characterization of endo-β-1,4-mannanase from Aspergillus fumigatus in Aspergillus sojae and Pichia pastoris. Biotechnol Prog 25:271–276

    Article  CAS  PubMed  Google Scholar 

  • Gilbert HJ, Stålbrand H, Brumer H (2008) How the walls come crumbling down: recent structural biochemistry of plant polysaccharide degradation. Curr Opin Plant Biol 11:338–348

    Article  CAS  PubMed  Google Scholar 

  • Grütter MG, Hawkes RB, Matthews BW (1979) Molecular basis of thermostability in the lysozyme from bacteriophage T4. Nature 277:667–669

    Article  PubMed  Google Scholar 

  • He X, Liua N, Li W, Zhang Z, Zhanga B, Ma Y (2008) Inducible and constitutive expression of a novel thermostable alkaline β-mannanase from alkaliphilic Bacillus sp. N16-5 in Pichia pastoris and characterization of the recombinant enzyme. Enzym Microb Technol 43:13–18

    Article  CAS  Google Scholar 

  • Heck JX, Soares LHB, Ayub MAZ (2005) Optimization of xylanase and mannanase production by Bacillus circulans strain BL53 on solid-state cultivation. Enzym Microb Technol 37:417–423

    Article  CAS  Google Scholar 

  • Hsiao YM, Liu YF, Fang MC, Tseng YH (2012) Transcriptional regulation and molecular characterization of the manA gene encoding the biofilm dispersing enzyme mannan endo-1,4-β-mannosidase in Xanthomonas campestris. J Agric Food Chem 58:1653–1663

    Article  Google Scholar 

  • Jin Y, Petricevic M, John A, Raich L, Jenkins H, Portela L, Souza D, Cuskin F, Gilbert HJ, Rovira C, Goddard-Borger ED, Williams SJ, Davies GJ (2016) A β-mannanase with a lysozyme-like fold and a novel molecular catalytic mechanism. ACS Cent Sci. doi:10.1021/acscentsci.6b00232

    PubMed  PubMed Central  Google Scholar 

  • Johnvesly B, Manjunath BR, Naik GR (2002) Pigeon pea waste as a novel, inexpensive, substrate for production of a thermostable alkaline protease from thermoalkalophilic Bacillus sp. JB-99. Bioresour Technol 82:61–64

    Article  CAS  PubMed  Google Scholar 

  • Katrolia P, Yan Q, Zhang P, Zhou P, Yang S, Jiang Z (2013) Gene cloning and enzymatic characterization of an alkali-tolerant endo-1,4-β-mannanase from Rhizomucor miehei. J Agric Food Chem 61:394–401

    Article  CAS  PubMed  Google Scholar 

  • Katrolia P, Zhou P, Zhang P, Yan Q, Li Y, Jiang Z, Xu H (2012) High level expression of a novel β-mannanase from Chaetomium sp. exhibiting efficient mannan hydrolysis. Carbohydr Polym 87:480–490

    Article  CAS  Google Scholar 

  • Liao H, Li S, Zheng H, Wei Z, Liu D, Raza W, Shen Q, Xu Y (2014) A new acidophilic thermostable endo-1,4-β-mannanase from Penicillium oxalicum GZ-2: cloning, characterization and functional expression in Pichia pastoris. BMC Biotechnol 14:90

    Article  PubMed  PubMed Central  Google Scholar 

  • Lu H, Luo H, Shi P, Huang H, Meng K, Yang P, Yao B (2014) A novel thermophilic endo-β-1,4-mannanase from Aspergillus nidulans XZ3: functional roles of carbohydrate-binding module and Thr/Ser-rich linker region. Appl Microbiol Biotechnol 98:2155–2163

    Article  CAS  PubMed  Google Scholar 

  • Lu H, Zhang H, Shi P, Luo H, Wang Y, Yang P, Yao B (2013) A family 5 β-mannanase from the thermophilic fungus Thielavia arenaria XZ7 with typical thermophilic enzyme features. Appl Microbiol Biotechnol 97:121–128

    Google Scholar 

  • Luo H, Wang K, Huang H, Shi P, Yang P, Yao B (2012) Gene cloning, expression, and biochemical characterization of an alkali-tolerant β-mannanase from Humicola insolens Y1. J Ind Microbiol Biotechnol 39:547–555

    Article  CAS  PubMed  Google Scholar 

  • Maijala P, Kango N, Szijarto N, Viikari L (2012) Characterization of hemicellulases from thermophilic fungi. Antonie Van Leeuwenhoek 101:905–917

    Article  CAS  PubMed  Google Scholar 

  • Martínez-Caballero S, Cano-Sánchez P, Mares-Mejía I, Díaz-Sánchez AG, Macías-Rubalcava ML, Hermoso JA, Rodríguez-Romero A (2014) Comparative study of two GH19 chitinase-like proteins from Hevea brasiliensis, one exhibiting a novel carbohydrate-binding domain. FEBS J 281:4535–4554

    Article  PubMed  Google Scholar 

  • Mikkelson A, Maaheimo H, Hakala TK (2013) Hydrolysis of konjac glucomannan by Trichoderma reesei mannanase and endoglucanases Cel7B and Cel5A for the production of glucomannooligosaccharides. Carbohydr Res 372:60–68

    Article  CAS  PubMed  Google Scholar 

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

    Article  CAS  Google Scholar 

  • Moreira LR, Filho EX (2008) An overview of mannan structure and mannan-degrading enzyme systems: a review. Appl Microbiol Biotechnol 79:165–178

    Article  CAS  PubMed  Google Scholar 

  • Nunes FM, Reis A, Domingues MR, Coimbra MA (2006) Characterization of galactomannan derivatives in roasted coffee beverages. J Agric Food Chem 54:3428–3439

    Article  CAS  PubMed  Google Scholar 

  • Pawar PM, Koutaniemi S, Tenkanen M, Mellerowicz EJ (2013) Acetylation of woody lignocellulose: significance and regulation. Front Plant Sci 4:118

    Article  PubMed  PubMed Central  Google Scholar 

  • Pérez RM, Carlucci MJ, Noseda MD, Matulewicz MC (2012) Chemical modifications of algal mannans and xylomannans: effects on antiviral activity. Phytochemistry 73:57–64

    Article  Google Scholar 

  • Puls J, Schuseil J (1993) In: Coughlan MP, Hazlewood GP (ed) Chemistry of hemicellulose: relationship between hemicellulose structure and enzyme required for hydrolysis. In Hemicellulose and Hemicellulases, London, Portland Press, pp 1–27

  • Scheller HV, Ulvskov P (2010) Hemicelluloses. Annu Rev Plant Biol 61:263–289

    Article  CAS  PubMed  Google Scholar 

  • Schröder R, Atkinson RG, Redgwell RJ (2009) Re-interpreting the role of endo-β-mannanases as mannan endo transglycosylase/hydrolases in the plant cell wall. Ann Bot 104:197–204

    Article  PubMed  PubMed Central  Google Scholar 

  • Shallom D, Shoham Y (2003) Microbial hemicellulases. Curr Opin Microbiol 6:219–228

    Article  CAS  PubMed  Google Scholar 

  • Shih P, Holland DR, Kirsch JF (1995) Thermal stability determinants of chicken egg-white lysozyme core mutants: hydrophobicity, packing volume, and conserved buried water molecules. Protein Sci 4:2050–2062

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Shimizu M, Fujii T, Masuo S, Takaya N (2010) Mechanism of de novo branched-chain amino acid synthesis as an alternative electron sink in hypoxic Aspergillus nidulans cells. Appl Environ Microbiol 76:1507–1515

  • Shimizu M, Kaneko Y, Ishihara S, Mochizuki M, Sakai K, Yamada M, Murata S, Itoh E, Yamamoto T, Sugiyama Y, Hirano T, Takaya N, Kobayashi T, Kato M (2015) Novel β-1,4-mannanase belonging to a new glycoside hydrolase family in Aspergillus nidulans. J Biol Chem 290:27914–27927

    CAS  PubMed  PubMed Central  Google Scholar 

  • Shimizu M, Masuo S, Fujita T, Doi Y, Kamimura Y, Takaya N (2012) Hydrolase controls cellular NAD, sirtuin, and secondary metabolites. Mol Cell Biol 32:3743–3755

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Shimizu M, Masuo S, Itoh E, Zhou S, Kato M, Takaya N (2016) Thiamine synthesis regulates the fermentation mechanisms in the fungus Aspergillus nidulans. Biosci Biotechnol Biochem 11:1–8

    Google Scholar 

  • Simkovic I (2013) Unexplored possibilities of all polysaccharide composites. Carbohydr Polm 95:697–715

    Article  CAS  Google Scholar 

  • Sims RE, Mabee W, Saddler JN, Taylor M (2010) An overview of second generation biofuel technologies: a review. Bioresour Technol 101:1570–1580

    Article  CAS  PubMed  Google Scholar 

  • Spencer DB, Chen CP, Hulett FM (1981) Effect of cobalt on synthesis and activation of Bacillus licheniformis alkaline phosphatase. J Bacteriol 145:926–933

    CAS  PubMed  PubMed Central  Google Scholar 

  • Stålbrand H, Siika-aho M, Tenkanen M, Viikari L (1993) Purification and characterization of two β-mannanases from Trichoderma reesei. J Biotechnol 29:229–242

    Article  Google Scholar 

  • Teleman A, Nordström M, Tenkanen M, Jacobs A, Dahlman O (2003) Isolation and characterization of O-acetylated glucomannans from aspen and birch wood. Carbohydr Res 338:525–534

    Article  CAS  PubMed  Google Scholar 

  • Timell TE (1967) Recent progress in the chemistry of wood hemicelluloses. Wood Sci Tech 1:45–70

    Article  CAS  Google Scholar 

  • Turner P, Mamo G, Karlsson EN (2007) Potential and utilization of thermophiles and thermostable enzymes in biorefining. Microb Cell Factories 6:9

    Article  Google Scholar 

  • Viikari L, Alapuranen M, Puranen T, Vehmaanper J, Siika-Aho M (2007) Thermostable enzymes in lignocellulose hydrolysis. Adv Biochem Eng Biotechnol 108:121–145

    CAS  PubMed  Google Scholar 

  • von Freiesleben P, Spodsberg N, Blicher TH, Anderson L, Jørgensen H, Stålbrand H, Meyer AS, Krogh KB (2016) An Aspergillus nidulans GH26 endo-β-mannanase with a novel degradation pattern on highly substituted galactomannans. Enzym Microb Technol 83:68–77

    Article  Google Scholar 

  • Wang C, Luo H, Niu C, Shi P, Huang H, Meng K, Bai Y, Wang K, Hua H, 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  PubMed  Google Scholar 

  • Wang C, Zhang J, Wang Y, Niu C, Ma R, Wang Y, Bai Y, Luo H, Yao B (2016) Biochemical characterization of an acidophilic β-mannanase from Gloeophyllum trabeum CBS900.73 with significant transglycosylation activity and feed digesting ability. Food Chem 197:474–481

    Article  CAS  PubMed  Google Scholar 

  • Yamabhai M, Ubol SS, Srila W, Haltrich D (2014) Mannan biotechnology: from biofuels to health. Crit Rev Biotechnol 36:32–42

    Article  Google Scholar 

  • Yoon KH, Chung S, Lim BL (2008) Characterization of the Bacillus subtilis WL-3 mannanase from a recombinant Escherichia coli. Microbiology 46:344–349

    CAS  Google Scholar 

  • Zhao W, Zheng J, Zhou H (2011a) A thermotolerant and cold-active mannan endo-1,4-β-mannosidase from Aspergillus niger CBS513.88: constitutive overexpression and high-density fermentation in Pichia pastoris. Bioresour Technol 102:7538–7547

    Article  CAS  PubMed  Google Scholar 

  • Zhao Y, Zhang Y, Cao Y, Qi J, Mao L, Xue Y, Gao F, Peng H, Wang X, Gao GF, Ma Y (2011b) Structural analysis of alkaline β-mannanase from Alkaliphilic Bacillus sp. N16-5: implications for adaptation to alkaline conditions. PLoS One 6:e14608–e14619

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgements

We thank Norma Foster for critical reading of the manuscript. This study was supported by a Grant-in-Aid for Scientific Research (20423535 to MS, 25450116 to MK). This study was also partially supported by the Institute for Fermentation, Osaka (IFO), Japan.

Author information

Authors and Affiliations

  1. Faculty of Agriculture, Meijo University, Nagoya, Aichi, 468-8502, Japan

    Kiyota Sakai, Mai Mochizuki, Miyuki Yamada, Yuta Shinzawa, Miho Minezawa, Saran Kimoto, Shunsuke Murata, Yuhei Kaneko, Saaya Ishihara, Masashi Kato & Motoyuki Shimizu

  2. Faculty of Science and Technology, Meijo University, Nagoya, Aichi, 468-8502, Japan

    Sadanari Jindou

  3. Graduate School of Bioagricultural Sciences, Nagoya University, Nagoya, Aichi, 464-8601, Japan

    Tetsuo Kobayashi

Authors
  1. Kiyota Sakai
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Mai Mochizuki
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Miyuki Yamada
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Yuta Shinzawa
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Miho Minezawa
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Saran Kimoto
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Shunsuke Murata
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Yuhei Kaneko
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Saaya Ishihara
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Sadanari Jindou
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Tetsuo Kobayashi
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. Masashi Kato
    View author publications

    You can also search for this author in PubMed Google Scholar

  13. Motoyuki Shimizu
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Motoyuki Shimizu.

Ethics declarations

Conflict of interest

The authors have no conflict of interest to declare.

Ethical approval

This article does not contain any studies with humans or other animals performed by any of the authors.

Electronic supplementary material

Fig. S1

(PDF 151 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Sakai, K., Mochizuki, M., Yamada, M. et al. Biochemical characterization of thermostable β-1,4-mannanase belonging to the glycoside hydrolase family 134 from Aspergillus oryzae . Appl Microbiol Biotechnol 101, 3237–3245 (2017). https://doi.org/10.1007/s00253-017-8107-x

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00253-017-8107-x

Keywords

Search

Navigation