Abstract
Thermo alkaline mannanase was purified from the bacteria of Bacillus pumilus (M27) using the techniques of ammonium sulphate precipitation, DEAE-Sephadex ion exchange chromatography and Sephacryl S200 gel filtration chromatography with 111-fold and 36 % yield. It was determined that the enzyme had 2 sub-units including 35 kDa and 55 kDa in gel filtration chromatography and SDS-PAGE electrophoresis systems. The optimum pH and temperature was determined as 8 and 60 °C, respectively. It was also noticed that the enzyme did not lose its activity at a wide interval such as pH 3–11 and at high temperatures such as 90 °C. Additionally, the effects of some metal ions on the mannanase enzyme activity. Moreover, the clarifying efficiency of purified mannanase enzyme with some fruit juices such as orange, apricot, grape and apple was also investigated. Enzymatic treatment was carried out with 1 mL L−1 of purified mannanase for 1 h at 60 °C. It was determined that the highest pure enzyme was efficient upon clarifying the apple juice at 154 % rate.
Similar content being viewed by others
References
Ademark P, Varga A, Medve J, Harjunpää V, Drakenberg T, Tjerneld F, Stålbrand H (1998) Softwood hemicellulose-degrading enzymes from Aspergillus niger: purification and properties of a beta-mannanase. J Biotech 63:199–210
Adiguzel G. (2008) Phenotypic and genotypic characterization of lactic acid bacteria isolated from Turkish dry fermente sausage PhD Thesis, Ataturk University, Graduate School of Health Sciences, Erzurum, Turkey, 110
Adiguzel G, Atasever M (2009) Phenotypic and genotypic characterization of lactic acid bacteria isolated from Turkish dry fermented sausage. Rom Biotech Lett 14(1):4130–4138
Adiguzel A, Inan K, Sahin F, Arasoglu T, Gulluce M, Belduz AO, Baris O (2011) Molecular diversity of thermophilic bacteria isolated from pasinler hot spring (Erzurum, Turkey). Turk J Biol 35:267–274
Adiguzel A, Ozkan H, Baris O, Inan K, Gulluce M (2009) Identification and characterization of thermophilic bacteria isolated from hot springs in Turkey. J Microbiol Meth 79:321–328
Akino T, Kato C, Horikoshi K (1989) Two Bacillus beta-mannanases having different COOH termini are produced in Escherichia coli carrying pMAH5. Appl Environ Microbiol 55(12):3178–3183
Altschul SF, Gish W, Miller W, Myers EW, Lipman DJ (1990) Basic local alignment search tool. J Mol Biol 215:403–410
Antara NS, Sujaya IN, Yokota A, Asano K, Aryanta WR, Tomita F (2002) Identification and succession of lacticacid bacteria during fermentation of ‘urutan’, a Balinese indigenous fermented sausage. World J Microbiol Biotech 18:255–262
Bailey MJ, Biely P, Poutanen K (1992) Interlaboratory testing methods for assay of xylanase activity. J Biotech 23:257–270
Beffa T, Blanc M, Lyon PF, Vogt G, Marchiani M, Fischer JL, Aragno M (1996) Isolation of Thermus strains from hot composts (60 to 80 °C). Appl Environ Microbiol 62:1723–1727
Bettiol JP, Showell MS (2002) US Patent 6376445
Bottone EJ, Peluso R (2003) Production by Bacillus pumilus (MSH) of an antifungal compound that is active against Mucoraceae and Aspergillus species: preliminary. J Med Microbiol 52:69–74
Bradford MM (1976) Rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Chem 2:248–253
Charrier M, Rouland C (2001) Mannan-degrading enzymes purified from the crop of the brown garden snail helix aspersa müller (Gastropoda pulmonata). J Exp Zool 290(2):125–135
Christgau S, Kauppinen S, Vind J, Kofod LV, Dalbøge H (1994) Expression cloning, purification and characterization of a beta-1,4-mannanase from Aspergillus aculeatus. Biochem Mol Biol Int l33(5):917–925
Civas A, Eberhard R, Le Dizet P, Petek F (1984) Glycosidases induced in Aspergillus tamarii Secreted alpha-D-galactosidase and beta-D-mannanase. Biochem J 219:857–863
Demir N, Nadaroglu H, Tasgin E, Adıgüzel A, Gulluce M (2011) Purification and characterization of a pectin lyase produced by Geobacillus stearothermophilus (Ah22) and fruit juice application. Ann Microbiol 61(4):939–946
Dhawan S, Kaur J (2007) Microbial mannanases: an overview of production and applications. Crit Rev Biotech 27(4):197–216
Drobniewski FA (1993) Bacillus cereus and related species. Clin Microbiol Rev 6:324–338
Filichkin SA, Priest HD, Givan SA, Shen R, Bryant DW, Fox SE, Wong WK, Mockler TC (2009) Genome-wide mapping of alternative splicing in Arabidopsis thaliana. Genome Res 20(1):45–58
Gibbs MD, Reeves RA, Sunna A, Bergquist L (1999) Sequencing and expression of a β-Mannnase gene from the extreme thermophilic Dictyoglomus thermophilum Rt46B1, and characteristics of the recombinant enzyme. Curr Microbiol 39:351–357
Halstead JR, Vercoe PE, Gilbert HJ, Davidson K, Hazlewood GP (1999) A family mannanase produced by Clostridium thermocellum as a component of the cellulosome contains a domain which is conserved in mannanases from anaerobic fungi. Microbiol 145:3101–3108
Holt JG, Noel RK (1984) Bergey’s Manual of Systematic Bacteriology, Volume 1–4. Lippincott Williams & Wilkins, New York
Kurakake M, Komaki T (2001) Production of β-mannanase and β-mannosidase from Aspergillus awamori K4 and their properties. Curr Microbiol 42:377–380
Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage. Nat 22:680–685
Liu Y, Lai Q, Dong C, Sun F, Wang L, Li G, Shao Z (2013) Phylogenetic diversity of the Bacillus pumilus group and the marine ecotype revealed by multilocus sequence analysis. Plos One 8(11):1–11. doi:10.1371/journal.pone0080097
Mandal V, Sen SK, Mandal NC (2008) Optimized culture conditions for bacteriocin production by Pediococcus acidilactici LAB 5 and its characterization. Indian J Biochem Biotech 45(2):106–110
Matarante A, Baruzzi F, Cocconcelli PS, Morea M (2004) Genotyping and toxigenic potential of Bacillus subtilis and Bacillus pumilus strains occurring in industrial and artisanal cured sausages. Appl Environ Microbiol 70(3):5168–5176
Matos JST, Jensen BB, Barreto SFHA, Hojberg O (2006) Spoilage and pathogenic bacteria ısolated from two types of Portuguese dry smoked sausages after shelf-life period in modified atmosphere package. Cienc Tecnol Aliment 5(3):165–174
Mendoza NS, Arai M, Kawaguchi T, Cubol FS, Panerio EG, Yoshida T (1994) Isolation of mannanutilizing bacteria and the culture conditions for mannanase production. World J Microbiol Biotechnol 10:51–54
Miller GL (1959) Use of dinitrosalicylic acid reagent for determination of reducing sugar. Anal Chem 31:426–428
Nadaroglu H, Demir N (2012) Purification and characterization of a novel serine protease compositain from compositae (Scorzonera hispanica L). Eur Food Res Technol 234(6):945–953
Nadaroglu H, Taskin E, Adiguzel A, Gulluce M, Demir N (2010) Production of a novel pectin lyase from bacillus pumilus (P9), purification and characterization and fruit juice application. Rom Biotech Lett 15(2):5167–5176
Parvathi A, Krishna K, Jose J, Joseph N, Nair S (2009) Biochemical and molecular characterization of Bacillus pumilus isolated from coastal environment in cochin, India. Braz J Microbiol 40:269–275
Pason P, Kyu KL, Ratanakhanokchai K (2006) Paenibacillus curdlanolyticus strain B-6 xylanolytic-cellulolytic enzyme system that degrades insoluble polysaccharides. Appl Environ Microbiol 72:2483–2490B
Setati ME, Ademark P, Van Zyl WH, Hahn-Hagerdal B, Stålbrand H (2001) Expression of the Aspergillus aculeatus Endo-b-1,4-mannanase Encoding Gene (man1) in Saccharomyces cerevisiae and characterization of the recombinant enzyme. Protein Exp Purif 21:105–114
Talbot G, Sygusch J (1990) Purification and characterization of thermostable beta- mannanase and alpha-galactosidase from Bacillus stearothermophilus. Appl Environ Microbiol 56(11):3505–3510
Tang CM, Waterman LD, Smith MH, Thurston CF (2001) The cel4 gene of Agaricus bisporus encodes a beta-mannanase. Appl Environ Microbiol 67(5):2298–2303
Tenkanen M, Makkonen M, Perttula M, Viikari L, Teleman A (1997) Action of Trichoderma reesei mannanase on galactoglucomannan in pine kraft pulp. J Biotechnol 57:191–204
Whitaker JR (1963) Determination of molecular weight of proteins by gel filtration on sephadex. Anal Chem 35:1950–1953
Yamamura I, Nozaki Y, Matsumoto T, Kato T (1996) Purification and some properties of endo-β-1,4 mannanase from a marine mollusk Littorina brevicula. Biosci Biotechnol Biochem 60:674–676
Yamanaka H, Moriyoshi K, Ohmoto T, Ohe T, Sakai K (2007) Degradation of bisphenol a by Bacillus pumilus isolated from kimchi, a traditionally fermented food. Appl Biochem Biotech 136:39–51
Zakaria MM, Ashiuchi M, Yamamoto S, Yagi T (1998) Optimization for β-mannanase production of a psychrophilic bacterium, Flavobacterium sp. Biosci Biotechnol Biochem 62:655–660
Zhao J, Shi P, Luo H, Yang P, Zhao H, Bai Y, Huang H, Wang H, Yao B (2010) An acidophilic and acid-stable β-mannanase from Phialophora sp P13 with high-mannan hydrolysis activity under simulated gastric conditions. J Agr Food Chem 58:3184–3190
Author information
Authors and Affiliations
Corresponding authors
Rights and permissions
About this article
Cite this article
Adiguzel, A., Nadaroglu, H. & Adiguzel, G. Purification and characterization of \( \boldsymbol{\upbeta} \)-mannanase from Bacillus pumilus (M27) and its applications in some fruit juices. J Food Sci Technol 52, 5292–5298 (2015). https://doi.org/10.1007/s13197-014-1609-y
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s13197-014-1609-y