Advertisement

Biotechnology Letters

, Volume 40, Issue 9–10, pp 1377–1387 | Cite as

Characterization of an inhibitor-resistant endo-1,4-β-mannanase from the gut microflora metagenome of Hermetia illucens

  • Jaeeun Song
  • Su-Yeon Kim
  • Dae-Hyuk Kim
  • Young-Seok Lee
  • Joon-Soo Sim
  • Bum-Soo Hahn
  • Chang-Muk Lee
Original Research Paper

Abstract

Objective

Hermetia illucens is a voracious insect scavenger that efficiently decomposes food waste. To exploit novel hydrolytic enzymes from this insect, we constructed a fosmid metagenome library using unculturable H. illucens intestinal microorganisms.

Results

Functional screening of the library on carboxymethyl cellulose plates identified a fosmid clone with a product displaying hydrolytic activity. Fosmid sequence analysis revealed a novel mannan-degrading gene (ManEM17) composed of 1371 base pairs, encoding 456 amino acids with a deduced 54 amino acid N-terminal signal peptide sequence. Conceptual translation and domain analysis revealed that sequence homology was highest (46%) with endo-1,4-β-mannosidase of Anaerophaga thermohalophila. Phylogenetic and domain analysis indicated that ManEM17 belongs to a novel β-mannanase containing a glycoside hydrolase family 26 domain. The recombinant protein (rManEM17) was expressed in Escherichia coli, exhibiting the highest activity at 55 °C and pH 6.5. The protein hydrolyzed substrates with β-1,4-glycosidic mannoses; maximum specific activity (5467 U mg−1) occurred toward locust bean gum galactomannan. However, rManEM17 did not hydrolyze p-Nitrophenyl-β-pyranosides, demonstrating endo-form mannanase activity. Furthermore, rManEM17 was highly stable under stringent conditions, including polar organic solvents as well as chemical reducing and denaturing reagents.

Conclusions

ManEM17 is an attractive candidate for mannan degradation under the high-organic-solvent and protein-denaturing processes in food and feed industries.

Keywords

Hermetia illucens Inhibitor resistance Larvae gut Mannanase Metagenome 

Notes

Acknowledgements

This work was supported by Grants from the National Institute of Agricultural Sciences, Rural Development Administration, Republic of Korea (Project No. PJ01045703 and PJ01086901).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

References

  1. 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:D233–D238.  https://doi.org/10.1093/nar/gkn663 CrossRefPubMedPubMedCentralGoogle Scholar
  2. Chauhan PS, Puri N, Sharma P, Gupta N (2012) Mannanases: microbial sources, production, properties and potential biotechnological applications. Appl Microbiol Biotechnol 93:1817–1830.  https://doi.org/10.1007/s00253-012-3887-5 CrossRefPubMedGoogle Scholar
  3. Cickova H, Newton GL, Lacy RC, Kozanek M (2015) The use of fly larvae for organic waste treatment. Waste Manag 35:68–80.  https://doi.org/10.1016/j.wasman.2014.09.026 CrossRefPubMedGoogle Scholar
  4. Ergun BG, Calik P (2016) Lignocellulose degrading extremozymes produced by Pichia pastoris: current status and future prospects. Bioproc Biosyst Eng 39:1–36.  https://doi.org/10.1007/s00449-015-1476-6 CrossRefGoogle Scholar
  5. Ferrer M, Martinez-Martinez M, Bargiela R, Streit WR, Golyshina OV, Golyshin PN (2016) Estimating the success of enzyme bioprospecting through metagenomics: current status and future trends. Microb Biotechnol 9:22–34.  https://doi.org/10.1111/1751-7915.12309 CrossRefPubMedGoogle Scholar
  6. Gibson LJ (2012) The hierarchical structure and mechanics of plant materials. J R Soc Interface 9:2749–2766.  https://doi.org/10.1098/rsif.2012.0341 CrossRefPubMedPubMedCentralGoogle Scholar
  7. Hatada Y et al (2005) Sequence of the gene for a high-alkaline mannanase from an alkaliphilic Bacillus sp. strain JAMB-750, its expression in Bacillus subtilis and characterization of the recombinant enzyme. Extremophiles 9:497–500.  https://doi.org/10.1007/s00792-005-0460-5 CrossRefPubMedGoogle Scholar
  8. Henrissat B (1991) A classification of glycosyl hydrolases based on amino-acid-sequence similarities. Biochem J 280:309–316CrossRefPubMedPubMedCentralGoogle Scholar
  9. Herve C, Rogowski A, Blake AW, Marcus SE, Gilbert HJ, Knox JP (2010) Carbohydrate-binding modules promote the enzymatic deconstruction of intact plant cell walls by targeting and proximity effects. Proc Natl Acad Sci USA 107:15293–15298.  https://doi.org/10.1073/pnas.1005732107 CrossRefPubMedGoogle Scholar
  10. Hogg D, Woo EJ, Bolam DN, McKie VA, Gilbert HJ, Pickersgill RW (2001) Crystal structure of mannanase 26A from Pseudomonas cellulosa and analysis of residues involved in substrate binding. J Biol Chem 276:31186–31192.  https://doi.org/10.1074/jbc.M010290200 CrossRefPubMedGoogle Scholar
  11. Inoue H, Yano S, Sawayama S (2015) Effect of beta-mannanase and beta-mannosidase supplementation on the total hydrolysis of softwood polysaccharides by the Talaromyces cellulolyticus cellulase system. Appl Biochem Biotech 176:1673–1686.  https://doi.org/10.1007/s12010-015-1669-0 CrossRefGoogle Scholar
  12. Ito N et al (1991) Novel thioether bond revealed by a 1.7-a crystal-structure of galactose-oxidase. Nature 350:87–90.  https://doi.org/10.1038/350087a0 CrossRefPubMedGoogle Scholar
  13. Jeon H, Park S, Choi J, Jeong G, Lee SB, Choi Y, Lee SJ (2011) The intestinal bacterial community in the food waste-reducing larvae of Hermetia illucens. Curr Microbiol 62:1390–1399.  https://doi.org/10.1007/s00284-011-9874-8 CrossRefPubMedGoogle Scholar
  14. Kaira GS, Panwar D, Kapoor M (2016) Recombinant endo-mannanase (ManB-1601) production using agro-industrial residues: development of economical medium and application in oil extraction from copra. Bioresour Technol 209:220–227.  https://doi.org/10.1016/j.biortech.2016.02.133 CrossRefPubMedGoogle Scholar
  15. Katsimpouras C, Dimarogona M, Petropoulos P, Christakopoulos P, Topakas E (2016) A thermostable GH26 endo-beta-mannanase from Myceliophthora thermophila capable of enhancing lignocellulose degradation. Appl Microbiol Biotechnol 100:8385–8397.  https://doi.org/10.1007/s00253-016-7609-2 CrossRefPubMedGoogle Scholar
  16. Kim GB, Seo YM, Kim CH, Paik IK (2011) Effect of dietary prebiotic supplementation on the performance, intestinal microflora, and immune response of broilers. Poultry Sci 90:75–82.  https://doi.org/10.3382/ps.2010-00732 CrossRefGoogle Scholar
  17. Klibanov AM (2001) Improving enzymes by using them in organic solvents. Nature 409:241–246.  https://doi.org/10.1038/35051719 CrossRefPubMedGoogle Scholar
  18. Lee CM et al (2014) Screening and characterization of a novel cellulase gene from the gut microflora of Hermetia illucens using metagenomic library. J Microbiol Biotechnol 24:1196–1206.  https://doi.org/10.4014/jmb.1405.05001 CrossRefPubMedGoogle Scholar
  19. Lee YS et al (2016) Identification of a novel alkaline amylopullulanase from a gut metagenome of Hermetia illucens. Int J Biol Macromol 82:514–521.  https://doi.org/10.1016/j.ijbiomac.2015.10.067 CrossRefPubMedGoogle Scholar
  20. Liu HX et al (2015) Biochemical characterization and cloning of an endo-1,4-beta-mannanase from Bacillus subtilis YH12 with unusually broad substrate profile. Process Biochem 50:712–721.  https://doi.org/10.1016/j.procbio.2015.02.011 CrossRefGoogle Scholar
  21. Maruthamuthu M, Jimenez DJ, Stevens P, van Elsas JD (2016) A multi-substrate approach for functional metagenomics-based screening for (hemi)cellulases in two wheat straw-degrading microbial consortia unveils novel thermoalkaliphilic enzymes. BMC Genom 17:86.  https://doi.org/10.1186/s12864-016-2404-0 CrossRefGoogle Scholar
  22. Moreira LRS, Filho EXF (2008) An overview of mannan structure and mannan-degrading enzyme systems. Appl Microbiol Biotechnol 79:165–178.  https://doi.org/10.1007/s00253-008-1423-4 CrossRefPubMedGoogle Scholar
  23. Patel AB, Patel AK, Shah MP, Parikh IK, Joshi CG (2016) Isolation and characterization of novel multifunctional recombinant family 26 glycoside hydrolase from Mehsani buffalo rumen metagenome. Biotechnol Appl Biochem 63:257–265.  https://doi.org/10.1002/bab.1358 CrossRefPubMedGoogle Scholar
  24. Politz O, Krah M, Thomsen KK, Borriss R (2000) A highly thermostable endo-(1,4)-beta-mannanase from the marine bacterium Rhodothermus marinus. Appl Microbiol Biotechnol 53:715–721CrossRefPubMedGoogle Scholar
  25. Songsiriritthigul C, Buranabanyat B, Haltrich D, Yamabhai M (2010) Efficient recombinant expression and secretion of a thermostable GH26 mannan endo-1,4-beta-mannosidase from Bacillus licheniformis in Escherichia coli. Microb Cell Fact 9:20.  https://doi.org/10.1186/1475-2859-9-20 CrossRefPubMedPubMedCentralGoogle Scholar
  26. Srivastava PK, Kapoor M (2017) Production, properties, and applications of endo-beta-mannanases. Biotechnol Adv 35:1–19.  https://doi.org/10.1016/j.biotechadv.2016.11.001 CrossRefPubMedGoogle Scholar
  27. Tailford LE et al (2009) Understanding how diverse beta-mannanases recognize heterogeneous substrates. Biochemistry 48:7009–7018.  https://doi.org/10.1021/bi900515d CrossRefPubMedGoogle Scholar
  28. Tester R et al (2012) The use of konjac glucomannan hydrolysates to recover healthy microbiota in infected vaginas treated with an antifungal agent. Benef Microbes 3:61–66.  https://doi.org/10.3920/Bm2011.0021 CrossRefPubMedGoogle Scholar
  29. Tsukagoshi H et al (2014) Structural and biochemical analyses of glycoside hydrolase family 26 beta-Mannanase from a symbiotic protist of the termite reticulitermes speratus. J Biol Chem 289:10843–10852.  https://doi.org/10.1074/jbc.M114.555383 CrossRefPubMedPubMedCentralGoogle Scholar
  30. Ufarte L, Laville E, Duquesne S, Potocki-Veronese G (2015) Metagenomics for the discovery of pollutant degrading enzymes. Biotechnol Adv 33:1845–1854.  https://doi.org/10.1016/j.biotechadv.2015.10.009 CrossRefPubMedGoogle Scholar
  31. van Zyl WH, Rose SH, Trollope K, Gorgens JF (2010) Fungal beta-mannanases: Mannan hydrolysis, heterologous production and biotechnological applications. Process Biochem 45:1203–1213.  https://doi.org/10.1016/j.procbio.2010.05.011 CrossRefGoogle Scholar
  32. Wang Y, Azhar S, Gandini R, Divne C, Ezcurra I, Aspeborg H (2015) Biochemical characterization of the novel endo-beta-mannanase AtMan5-2 from Arabidopsis thaliana. Plant Sci 241:151–163.  https://doi.org/10.1016/j.plantsci.2015.10.002 CrossRefPubMedGoogle Scholar
  33. Yamabhai M, Sak-Ubol S, Srila W, Haltrich D (2016) Mannan biotechnology: from biofuels to health. Crit Rev Biotechnol 36:32–42.  https://doi.org/10.3109/07388551.2014.923372 CrossRefPubMedGoogle Scholar
  34. Zahura UA, Rahman MM, Inoue A, Tanaka H, Ojima T (2010) An endo-beta-1,4-mannanase, AkMan, from the common sea hare Aplysia kurodai. Comp Biochem Phys B 157:137–143.  https://doi.org/10.1016/j.cbpb.2010.05.012 CrossRefGoogle Scholar

Copyright information

© Springer Nature B.V. 2018

Authors and Affiliations

  1. 1.Metabolic Engineering DivisionNational Institute of Agricultural Sciences, Rural Development AdministrationJeonjuRepublic of Korea
  2. 2.Department of Molecular Biology, Institute for Molecular Biology and GeneticsChonbuk National UniversityJeonjuRepublic of Korea

Personalised recommendations