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Purification and Characterization of α-Mannosidase from Moss Hyophilla nymaniana (Fleish.) Menzel

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Proceedings of the National Academy of Sciences, India Section B: Biological Sciences Aims and scope Submit manuscript

Abstract

α-Mannosidase EC: (3.2.1.24) is the major post-translational modifications enzyme which catalyzes the cleavage of the alpha form of mannose. Lysosomal α-mannosidase assists in the processing of complex sugars from glycoproteins. Defective or deficient α-mannosidase activity causes α-mannosidosis and leads to deterioration of the central nervous system in children. In the present study, plant α-mannosidase was purified 39-fold in a stepwise manner by ammonium sulfate precipitation, dialysis, gel filtration chromatography and DEAE ion exchange chromatography. The purified mannosidase was a heterodimer consisting of approximately 72 and 50 kDa molecular mass subunits under reducing conditions. Optimum pH and temperature reported for mannosidase activity were 4.5 and 40 °C respectively. Among the various metal ions tested for the assessment of the enzyme activity, 2 mM Zn2+ strongly activated the enzyme while Hg2+and Ag2+ caused enzyme inhibition at the same concentration. The Km and Vmax values calculated from Lineweaver plot against the substrate p-nitrophenyl α-mannopyranoside were 5.4 mM and 8.4 U/ml, respectively. To the author’s knowledge, the data presented here is the first report on purification and characterization of α-mannosidase from the moss H. nymaniana.

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References

  1. Forsee WT, Palmer CF, Schutzbach JS (1989) Purification and characterization of an α-1,2-mannosidase involved in processing asparagines-linked oligosaccharides. J Biol Chem 264:3869–3876

    CAS  PubMed  Google Scholar 

  2. Hossain MA, Nakano R, Nakamura K, Hossain MT, Kimura Y (2010) Molecular characterization of plant acidic alpha-mannosidase, a member of glycosylhydrolase family 38, involved in the turnover of N-glycans during tomato fruit ripening. J Biochem 148(5):603–616

    Article  CAS  PubMed  Google Scholar 

  3. Pastuszak I, Kaushal GP, Wall KA, Pan YT, Sturm A, Elbein AD (1990) Purification and properties of arylmannosidases from mung bean seedlings and soybean cells. Glycobiol 1:71–82

    Article  CAS  Google Scholar 

  4. Woo KK, Kimura Y (2005) Regulation of substrate specificity of plant α-mannosidase by cobalt ion: in vitro hydrolysis of high-mannose type N-glycans by Co2+ activated Ginkgo α-mannosidase. Biosci Biotechnol Biochem 69:1111–1119

    Article  CAS  PubMed  Google Scholar 

  5. Moremen KW, Trimble RB, Herscovics A (1994) Glycosidases of the asparagines-linked oligosaccharide processing pathway. Glycobiology 4:113–125

    Article  CAS  PubMed  Google Scholar 

  6. Moremen K (2000) α-mannosidases in asparagine-linked oligosaccharide processing and catabolism. In: Ernst B, Hart G, Sinay P (eds) Oligosaccharides in chemistry and biology: a comprehensive handbook, vol II. Wiley, New York, pp 81–117

    Google Scholar 

  7. Moremen KW (2002) Golgi alpha-mannosidase II deficiency in vertebrate systems: implications for asparagine-linked oligosaccharide processing in mammals. Biochim Biophys Acta 1573:225–235

    Article  CAS  PubMed  Google Scholar 

  8. Henrissat B, Bairoch A (1993) New families in the classification of glycosyl hydrolases based on amino acid sequence similarities. Biochem J 293:781–788

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Christgau S, Andersen LN, Kauppinen S, Heldt-hansen HP, Dalboege H (1994) Purified enzyme exhibiting mannanase activity; application in oil, paper, pulp, fruit and vegetable juice industry and carrageenan extraction. Patent Novo-Nordisk 9425576:10

    Google Scholar 

  10. Akino T, Nakamura N, Horikoshi K (1988) Characterization of three α-mannanases of an alkalophylic Bacillus sp. Agric Biol Chem 52:773–779

    CAS  Google Scholar 

  11. Viikari L, Kantelinen A, Sundquist J, Linko M (1994) Xylanases in bleaching: from an idea to the industry. FEMS Microbiol Rev 13:335–350

    Article  CAS  Google Scholar 

  12. Lahtinen T, Kristoo P, Paloheimo M (1995). Mannanases in softwood kraft pulp bleaching. In: Abstract PG4-101, sixth international conference on biotechnology in the pulp and paper industry, 11–15 June 1995, Vienna, Austria

  13. Jacob GS, Scudder P (1994) Glycosidases in structural analysis. Methods Enzymol 230:280–299

    Article  CAS  PubMed  Google Scholar 

  14. Ajisaka K, Matsuo I, Isomura M, Fujimoto H, Shirakabe M, Okawa M (1995) Enzymatic synthesis of mannobioses and mannotrioses by reverse hydrolysis using alpha-mannosidase from Aspergillus niger. Carbohydr Res 270(2):123–130

    Article  CAS  PubMed  Google Scholar 

  15. Chin LH, Ali ZM, Lazan H (1999) Cell wall modifications, degrading enzymes and softening of 239 carambola fruit during ripening. J Exp Bot 50:767–775

    Article  CAS  Google Scholar 

  16. Sun H, Wofe JH (2001) Recent progress in lysosomal α-mannosidase and its deficiency. Exp Mol Med 33(1):1–17

    Article  CAS  PubMed  Google Scholar 

  17. Kestwal RM, Bhinde SV (2005) Purification and partial characterization of α-D-mannosidase from Erythrinaindica seeds. Indian J Biochem Biophys 42:156–160

    CAS  PubMed  Google Scholar 

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

    CAS  PubMed  Google Scholar 

  19. Tulasi BR, Nandimpalli KS (1999) Purification of mannosidase activity from Indian lablab beans. Biochem Mol Biol Int 41:925–931

    Google Scholar 

  20. Jagdeesh BH, Prabha TN, Srinivias K (2004) Activities of β-hexosaminidase and α-mannosidase during development and ripening of bell capsicum (Capsicum annuumvar. variata). Plant Sci 167:1263–1271

    Article  CAS  Google Scholar 

  21. Einhoff W, Rudiger H (1988) The alpha-mannosidase from Canavalia ensiformis seeds: chemical and kinetic properties and action on animal lymphocytes. Biol Chem Hoppe Syler 369(3):165–169

    Article  CAS  Google Scholar 

  22. Ghosh S, Meli VS, Kumar A, Thakur A, Chakaraborty N, Chakaraborty S, Dutta A (2011) The N-glycan processing enzymes alpha-mannosidase and beta-D-N-acetylhexosaminidase are involved in ripening-associated softening in the non-climacteric fruits of capsicum. J Exp Bot 62(2):571–582

    Article  CAS  PubMed  Google Scholar 

  23. Niyogi K, Singh M (1988) An immunological distinct form of alpha-mannosidase in Canavalia ensiformis leaf. Phytochemistry 27:2737–2741

    Article  CAS  Google Scholar 

  24. Conti S, Carratu G, Ginnatassio M (1987) α-D-Mannosidase from the aleurone layers of resting wheat grains: sugar-depleted forms. Phytochemistry 26:2909–2912

    Article  CAS  Google Scholar 

  25. Kestwal RM, Konozy EH, Hsiao CD, Roque-Barreira MC, Bhide SV (2007) Characterization of alpha-mannosidase from erythrina indica seeds and influence of endogenous lectin on its activity. Biochim Biophys Acta 1770:24–28

    Article  CAS  PubMed  Google Scholar 

  26. Woo KK, Miyazaki M, Hara S, Kimura M, Kimura Y (2014) Purification and characterization of Co (II) sensitive α mannosidase from Ginkgo biloba seeds. Biosci Biotechnol Biochem 68(12):2547–2556

    Article  Google Scholar 

  27. Nakagawa H, Enomoto N, Asakawa N, Uda Y (1988) Purification and characterization of α-mannosidase and β- N-acetylhexosaminidase from watermelon fruit. Agric Biol Chem 59(2):2223–2230

    Google Scholar 

  28. Vega R, Dominguez A (1987) Partial characterization of α-mannosidase from Yarrowia lipolytica. J Basic Microbiol 28(6):371–379

    Article  Google Scholar 

  29. Ahi AT, Gonnety JT, Faulet BM, Kouame LP, Nimake S (2007) Biochemical characterization of two α-mannosidases from breadfruit (Artocarpus communis) seeds. Afr J Biochem Res 1(6):106–116

    Google Scholar 

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Acknowledgements

The authors deeply acknowledge the support got from the Government from UGC Project F. No. 37-116/2009 (SR). They are also thankful to Birla Institute of Technology, Department of Bioengineering and Centre of Excellence (COE) TEQUIP II for providing R&D facilities.

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Authors and Affiliations

  1. Department of Bio-Engineering, Birla Institute of Technology, Mesra, Ranchi, 835215, India

    Rashmi Mishra & Ramesh Chandra

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  1. Rashmi Mishra
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  2. Ramesh Chandra
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Correspondence to Ramesh Chandra.

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The authors declare that there is no financial or commercial conflict of interest to publish this manuscript.

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Significance statement This is the first study documenting enzyme isolation, purification and characterization of α-mannosidase in lower plant mosses. This will help in development of in vitro glycosylation of desired protein in future and pave the way for engineered glycoprotein formation.

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Mishra, R., Chandra, R. Purification and Characterization of α-Mannosidase from Moss Hyophilla nymaniana (Fleish.) Menzel. Proc. Natl. Acad. Sci., India, Sect. B Biol. Sci. 89, 885–891 (2019). https://doi.org/10.1007/s40011-018-1001-1

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  • DOI: https://doi.org/10.1007/s40011-018-1001-1

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