Biochemical properties of a new thermo- and solvent-stable xylanase recovered using three phase partitioning from the extract of Bacillus oceanisediminis strain SJ3

Abstract

The present study investigates the production and partial biochemical characterization of an extracellular thermostable xylanase from the Bacillus oceanisediminis strain SJ3 newly recovered from Algerian soil using three phase partitioning (TPP). The maximum xylanase activity recorded after 2 days of incubation at 37 °C was 20.24 U/ml in the presence of oat spelt xylan. The results indicated that the enzyme recovered in the middle phase of TPP system using the optimum parameters were determined as 50% ammonium sulfate saturation with 1.0:1.5 ratio of crude extract: t-butanol at pH and temperature of 8.0 and 10 °C, respectively. The xylanase was recovered with 3.48 purification fold and 107% activity recovery. The enzyme was optimally active at pH 7.0 and was stable over a broad pH range of 5.0–10. The optimum temperature for xylanase activity was 55 °C and the half-life time at this temperature was of 6 h. At this time point the enzyme retained 50% of its activity after incubation for 2 h at 95 °C. The crude enzyme resist to sodium dodecyl sulfate and β-mercaptoethanol, while all the tested ions do not affect the activity of the enzyme. The recovered enzyme is, at least, stable in tested organic solvents except in propanol where a reduction of 46.5% was observed. Further, the stability of the xylanase was higher in hydrophobic solvents where a maximum stability was observed with cyclohexane. These properties make this enzyme to be highly thermostable and may be suggested as a potential candidate for application in some industrial processes. To the best of our knowledge, this is the first report of xylanase activity and recoverey using three phase partitioning from B. oceanisediminis.

Keywords

Bacillus oceanisediminis / Xylanase / Thermostability / Hydrophobic solvents / Industrial processes / Three phase partitioning

References

[1] Amore A, Parameswaran B, Kumar R, Birolo L, Vinciguerra R, Marcolongo L, Ionata E, La Cara F, Pandey A, Faraco V. Application of a new xylanase activity from Bacillus amyloliquefaciens XR44A in brewer’s spent grain saccharification. J Chem Technol Biotechnol, 2015, 90: 573-581.

[2] Badhan AK, Chadha BS, Kaur J, Saini HS, Bhat MK. Production of multiple xylanolytic and cellulolytic enzymes by thermophilic fungus Myceliophthora sp. IMI 387099. Bioresour Technol, 2007, 98: 504-510.

[3] Baek CU, Lee SG, Chung YR, Cho I, Kim JH. Cloning of a family 11 xylanase gene from Bacillus amyloliquefaciens CH51 isolated from Cheonggukjang. Indian J Microbiol, 2012, 52: 695-700.

[4] Bastawde KB. Xylan structure, microbial xylanases, and their mode of action. World J Microbiol Biotechnol, 1992, 8: 353-368.

[5] Bataillon M, Nunes Cardinali A, Castillon N, Duchiron F. Purification and characterization of a moderately thermostable xylanase from Bacillus sp. strain SPS-0. Enzyme Microb Technol, 2000, 26: 187-192.

[6] Blanco A, Vidal T, Colom JF, Pastor FI. Purification and properties of xylanase A from alkali-tolerant Bacillus sp. strain BP-23. Appl Environ Microbiol, 1995, 61: 4468-4470.

[7] Bouacem K, Bouanane-Darenfed A, Boucherba N, Joseph M, Gagaoua M, Ben Hania W, Kecha M, Benallaoua S, Hacene H, Ollivier B, Fardeau ML. Partial characterization of xylanase produced by Caldicoprobacter algeriensis, a new thermophilic anaerobic bacterium isolated from an Algerian hot spring. Appl Biochem Biotechnol, 2014, 174: 1969-1981.

[8] Bouanane-Darenfed A, Boucherba N, Bouacem K, Gagaoua M, Joseph M, Kebbouche-Gana S, Nateche F, Hacene H, Ollivier B, Cayol J-L, Fardeau M-L. Characterization of a purified thermostable xylanase from Caldicoprobacter algeriensis sp. nov. strain TH7C1T. Carbohyd Res, 2016, 419: 60-68.

[9] Boucherba N, Gagaoua M, Copinet E, Bettache A, Duchiron F, Benallaoua S. Purification and characterization of the xylanase produced by Jonesia denitrificans BN-13. Appl Biochem Biotechnol, 2014, 172: 2694-2705.

[10] Chang S, Guo Y, Wu B, He B. Extracellular expression of alkali tolerant xylanase from Bacillus subtilis Lucky9 in E. coli and application for xylooligosaccharides production from agro-industrial waste. Int J Biol Macromol, 2017, 96: 249-256.

[11] Collins T, Gerday C, Feller G. Xylanases, xylanase families and extremophilic xylanases. FEMS Microbiol Rev, 2005, 29: 3-23.

[12] Dheeran P, Nandhagopal N, Kumar S, Jaiswal YK, Adhikari DK. A novel thermostable xylanase of Paenibacillus macerans IIPSP3 isolated from the termite gut. J Ind Microbiol Biotechnol, 2012, 39(6): 851-860.

[13] Elgharbi F, Hlima HB, Farhat-Khemakhem A, Ayadi-Zouari D, Bejar S, Hmida-Sayari A. Expression of A. niger US368 xylanase in E. coli: purification, characterization and copper activation. Int J Biol Macromol, 2015, 74: 263-270.

[14] Elgharbi F, Hmida-Sayari A, Zaafouri Y, Bejar S. Expression of an Aspergillus niger xylanase in yeast: application in breadmaking and in vitro digestion. Int J Biol Macromol, 2015, 79: 103-109.

[15] Ellis JT, Magnuson TS (2012) Thermostable and alkalistable xylanases produced by the thermophilic bacterium Anoxybacillus flavithermus TWXYL3. ISRN microbiology

[16] Fujimoto H, Ooi T, Wang S-L, Takizawa T, Hidaka H, Murao S, Arai M. Purification and properties of three xylanases from Aspergillus aculeatus. Biosci Biotechnol Biochem, 1995, 59: 538-540.

[17] Gagaoua M, Hafid K. Three phase partitioning system, an emerging non-chromatographic tool for proteolytic enzymes recovery and purification. Biosens J, 2016, 5(1): 100134.

[18] Gagaoua M, Boucherba N, Bouanane-Darenfed A, Ziane F, Nait-Rabah S, Hafid K, Boudechicha HR. Three-phase partitioning as an efficient method for the purification and recovery of ficin from Mediterranean fig (Ficus carica L.) latex. Sep Purif Technol, 2014, 132: 461-467.

[19] Gagaoua M, Hoggas N, Hafid K. Three phase partitioning of zingibain, a milk-clotting enzyme from Zingiber officinale Roscoe rhizomes. Int J Biol Macromol, 2015, 73: 245-252.

[20] Gagaoua M, Hafid K, Hoggas N. Data in support of three phase partitioning of zingibain, a milk-clotting enzyme from Zingiber officinale Roscoe rhizomes. Data in brief, 2016, 6: 634-639.

[21] Gagaoua M, Ziane F, Nait Rabah S, Boucherba N, El-Hadef El-Okki Ait Kaki A, Bouanane-Darenfed A, Hafid K. Three phase partitioning, a scalable method for the purification and recovery of cucumisin, a milk-clotting enzyme, from the juice of Cucumis melo var. reticulatus. Int J Biol Macromol, 2017, 102: 515-525.

[22] Gang GJ, Zbijewski W, Webster Stayman J, Siewerdsen JH. Cascaded systems analysis of noise and detectability in dual-energy cone-beam CT. Med Phys, 2012, 39: 5145-5156.

[23] Garg G, Dhiman SS, Mahajan R, Kaur A, Sharma J. Bleach-boosting effect of crude xylanase from Bacillus stearothermophilus SDX on wheat straw pulp. N Biotechnol, 2011, 28: 58-64.

[24] Gaur R, Tiwari S. Isolation, production, purification and characterization of an organic-solvent-thermostable alkalophilic cellulase from Bacillus vallismortis RG-07. BMC Biotechnol, 2015, 15: 19.

[25] Gaur R, Tiwari S, Rai P, Srivastava V. Isolation, production, and characterization of thermotolerant xylanase from solvent tolerant Bacillus vallismortis RSPP-15. Int J Polym Sci, 2015, 2015: 10.

[26] Goswami GK, Pathak RR, Krishnamohan M, Ramesh B. Production, partial purification and biochemical characterization of thermostable xylanase from Bacillus brevis. Biomed Pharmacol J, 2013, 6: 435-440.

[27] Goswami GK, Krishnamohan M, Nain V, Aggarwal C, Ramesh B. Cloning and heterologous expression of cellulose free thermostable xylanase from Bacillus brevis. SpringerPlus, 2014, 3(1): 20.

[28] Gowdhaman D, Manaswini VS, Jayanthi V, Dhanasri M, Jeyalakshmi G, Gunasekar V, Sugumaran KR, Ponnusami V. Xylanase production from Bacillus aerophilus KGJ2 and its application in xylooligosaccharides preparation. Int J Biol Macromol, 2014, 64: 90-98.

[29] Guo G, Liu Z, Xu J, Liu J, Dai X, Xie D, Peng K, Feng X, Duan S, Zheng K, Cheng L, Fu Y. Purification and characterization of a xylanase from Bacillus subtilis isolated from the degumming line. J Basic Microbiol, 2012, 52: 419-428.

[30] Gurtler V, Stanisich VA. New approaches to typing and identification of bacteria using the 16S-23S rDNA spacer region. Micobiology, 1996, 142: 3-16.

[31] Hakulinen N, Turunen O, Janis J, Leisola M, Rouvinen J. Three-dimensional structures of thermophilic beta-1,4-xylanases from Chaetomium thermophilum and Nonomuraea flexuosa. Comparison of twelve xylanases in relation to their thermal stability. Eur J Biochem, 2003, 270: 1399-1412.

[32] Hmida-Sayari A, Taktek S, Elgharbi F, Bejar S. Biochemical characterization, cloning and molecular modeling of a detergent and organic solvent-stable family 11 xylanase from the newly isolated Aspergillus niger US368 strain. Process Biochem, 2012, 47: 1839-1847.

[33] Irfan M, Nadeem M, Syed Q, Baig S. Effect of medium composition on xylanase production by Bacillus subtilis using various agricultural wastes. Am Eurasian J Agric Environ Sci, 2012, 12: 561-565.

[34] Jain A, Krishnan KP. A glimpse of the diversity of complex polysaccharide-degrading culturable bacteria from Kongsfjorden, Arctic Ocean. Ann Microbiol, 2017, 2: 203-214.

[35] Kim OS, Cho YJ, Lee K, Yoon SH, Kim M, Na H, Park SC, Jeon YS, Lee JH, Yi H, Won S, Chun J. Introducing EzTaxon-e: a prokaryotic 16S rRNA gene sequence database with phylotypes that represent uncultured species. Int J Syst Evol Microbiol, 2012, 62: 716-721.

[36] Kumar V, Satyanarayana T. Biochemical and thermodynamic characteristics of thermo-alkali-stable xylanase from a novel polyextremophilic Bacillus halodurans TSEV1. Extremophiles, 2013, 17: 797-808.

[37] Kumar V, Satyanarayana T. Production of thermo-alkali-stable xylanase by a novel polyextremophilic Bacillus halodurans TSEV1 in cane molasses medium and its applicability in making whole wheat bread. Bioprocess Biosyst Eng, 2014, 37: 1043-1053.

[38] Kumar D, Verma R, Sharma P, Rana A, Sharma R, Prakash C, Bhalla TC. Production and partial purification of xylanase from a new thermophilic isolate. In Biol Forum Int J, 2010, 2: 83-87.

[39] Kumar L, Nagar S, Mittal A, Garg N, Gupta VK. Immobilization of xylanase purified from Bacillus pumilus VLK-1 and its application in enrichment of orange and grape juices. J Food Sci Technol, 2014, 51: 1737-1749.

[40] Lindner C, Stulke J, Hecker M. Regulation of xylanolytic enzymes in Bacillus subtilis. Microbiology, 1994, 140(Pt 4): 753-757.

[41] Logan NA, Berkeley RC. Identification of Bacillus strains using the API system. J Gen Microbiol, 1984, 130: 1871-1882.

[42] Lv Z, Yang J, Yuan H. Production, purification and characterization of an alkaliphilic endo-β-1,4-xylanase from a microbial community EMSD5. Enzyme Microb Technol, 2008, 43: 343-348.

[43] Maalej I, Belhaj I, Masmoudi NF, Belghith H. Highly thermostable xylanase of the thermophilic fungus Talaromyces thermophilus: purification and characterization. Appl Biochem Biotechnol, 2009, 158: 200-212.

[44] Mamo G, Hatti-Kaul R, Mattiasson B. A thermostable alkaline active endo-β-1-4-xylanase from Bacillus halodurans S7: purification and characterization. Enzyme Microb Technol, 2006, 39: 1492-1498.

[45] Miller GL. Use of dinitrosalycilic acid reagent for determination of reducing sugars. Anal Chem, 1959, 31: 426-428.

[46] Nagar S, Gupta VK, Kumar D, Kumar L, Kuhad RC. Production and optimization of cellulase-free, alkali-stable xylanase by Bacillus pumilus SV-85S in submerged fermentation. J Ind Microbiol Biotechnol, 2010, 37: 71-83.

[47] Nagar S, Mittal A, Kumar D, Gupta VK. Production of alkali tolerant cellulase free xylanase in high levels by Bacillus pumilus SV-205. Int J Biol Macromol, 2012, 50(2): 414-420.

[48] Nakamura S, Nakai R, Namba K, Kubo T, Wakabayashi K, Aono R, Horikoshi K. Structure-function relationship of the xylanase from alkaliphilic Bacillus sp. strain 41M-1. Nucleic Acids Symp Ser, 1995, 34: 99-100.

[49] Ozcan BD, Coskun A, Ozcan N, Baylan M. Some properties of a new thermostable xylanase from alkaliphilic and thermophilic Bacillus sp.isolate DM-15. J Anim Vet Adv, 2011, 10: 138-143.

[50] Perez-Rodriguez N, Oliveira F, Perez-Bibbins B, Belo I, Torrado Agrasar A, Dominguez JM. Optimization of xylanase production by filamentous fungi in solid-state fermentation and scale-up to horizontal tube bioreactor. Appl Biochem Biotechnol, 2014, 173: 803-825.

[51] Poosarla VG, Chandra TS. Purification and characterization of novel halo-acid-alkali-thermo-stable xylanase from Gracilibacillus sp. TSCPVG. Appl Biochem Biotechnol, 2014, 173(6): 1375-1390.

[52] Prakash P, Jayalakshmi SK, Prakash B, Rubul M, Sreeramulu K. Production of alkaliphilic, halotolerent, thermostable cellulase free xylanase by Bacillus halodurans PPKS-2 using agro waste: single step purification and characterization. World J Microbiol Biotechnol, 2012, 28: 183-192.

[53] Pratumteep A, Sansernsuk J, Nitisinprasert S, Apiraksakorn J. Production, characterization and hydrolysation products of xylanase from Bacillus subtilis GN156. KKU Res J, 2010, 15: 343-350.

[54] Roy I, Sharma A, Gupta MN. Three phase partitioning for simultaneous renaturation and partial purification of Aspergillus niger xylanase. BBA Proteins Proteom, 2004, 1698: 107-110.

[55] Roy I, Sharma A, Gupta MN. Recovery of biological activity in reversibly inactivated proteins by three phase partitioning. Enzyme Microb Technol, 2005, 37: 113-120.

[56] Sambrook J, Fritsch E, Maniatis T. Molecular cloning: a laboratory manual, 1989, 2, Cold Spring Harbor: Cold Spring Harbor Laboratory Press.

[57] Sanghi A, Garg N, Kuhar K, Kuhad RC, Gupta VK. Enhanced production of cellulase-free xylanase by alkalophilic Bacillus subtilis ASH and its application in biobleaching of kraft pulp. BioResources, 2009, 4: 1109-1129.

[58] Sanghi A, Garg N, Gupta VK, Mittal A, Kuhad RC. One-step purification and characterization of cellulase-free xylanase produced by alkalophilic Bacillus subtilis ash. Braz J Microbiol, 2010, 41: 467-476.

[59] Sardar M, Sharma A, Gupta MN. Refolding of a denatured α-chymotrypsin and its smart bioconjugate by three-phase partitioning. Biocatal Biotransform, 2007, 25: 92-97.

[60] Seo JK, Park TS, Kwon IH, Piao MY, Lee CH, Ha JK. Characterization of cellulolytic and xylanolytic enzymes of Bacillus licheniformis JK7 isolated from the rumen of a native Korean goat. Asian Australas J Anim Sci, 2013, 26: 50-58.

[61] Shameer S. Haloalkaliphilic Bacillus species from solar salterns: an ideal prokaryote for bioprospecting studies. Ann Microbiol, 2016, 66: 1315-1327.

[62] Sharma A, Gupta MN. Macroaffinity ligand-facilitated three-phase partitioning (MLFTPP) for purification of xylanase. Biotechnol Bioeng, 2002, 80: 228-232.

[63] Sharma M, Chadha BS, Kaur M, Ghatora SK, Saini HS. Molecular characterization of multiple xylanase producing thermophilic/thermotolerant fungi isolated from composting materials. Lett Appl Microbiol, 2008, 46: 526-535.

[64] Subramaniyan S. Isolation, purification and characterisation of low molecular weight xylanase from Bacillus pumilus SSP-34. Appl Biochem Biotechnol, 2012, 166(7): 1831-1842.

[65] Subramaniyan S, Prema P. Cellulase-free xylanases from Bacillus and other microorganisms. FEMS Microbiol Lett, 2000, 183: 1-7.

[66] Tarayre C, Brognaux A, Brasseur C, Bauwens J, Millet C, Matteotti C, Destain J, Vandenbol M, Portetelle D, De Pauw E, Haubruge E, Francis F, Thonart P. Isolation and cultivation of a xylanolytic Bacillus subtilis extracted from the gut of the termite Reticulitermes santonensis. Appl Biochem Biotechnol, 2013, 171: 225-245.

[67] Techapun C, Poosaran N, Watanabe M, Sasaki K. Optimization of aeration and agitation rates to improve cellulase-free xylanase production by thermotolerant Streptomyces sp. Ab106 and repeated fed-batch cultivation using agricultural waste. J Biosci Bioeng, 2003, 95: 298-301.

[68] Terrasan CR, Temer B, Duarte MC, Carmona EC. Production of xylanolytic enzymes by Penicillium janczewskii. Bioresour Technol, 2010, 101: 4139-4143.

[69] Trajano HL, Pattathil S, Tomkins BA, Tschaplinski TJ, Hahn MG, Van Berkel GJ, Wyman CE. Xylan hydrolysis in Populus trichocarpa × P. deltoides and model substrates during hydrothermal pretreatment. Bioresour Technol, 2014, 179C: 202-210.

[70] Viet DN, Kamio Y, Abe N, Kaneko J, Izaki K. Purification and properties of beta-1, 4-xylanase from Aeromonas caviae W-61. Appl Environ Microbiol, 1991, 57: 445-449.

[71] Wahyuntari B, Mubarik NR, Setyahadi S. Effect of pH, temperature and medium composition on xylanase production by Bacillus sp. AQ-1 and partial characterization of the crude enzyme. Microbiology, 2009, 3: 17-22.

[72] Wang W, Wang Z, Cheng B, Zhang J, Li C, Liu X, Yang C. High secretory production of an alkaliphilic actinomycete xylanase and functional roles of some important residues. World J Microbiol Biotechnol, 2014, 30: 2053-2062.

[73] Wong KK, Martin LA, Gama FM, Saddler JN, de Jong E. Bleach boosting and direct brightening by multiple xylanase treatments during peroxide bleaching of kraft pulps. Biotechnol Bioeng, 1997, 54: 312-318.

[74] Xie Z, Lin W, Luo J. Genome sequence of Cellvibrio pealriver PR1, a xylanolytic and agarolytic bacterium isolated from freshwater. J Biotechnol, 2015, 214: 57-58.

[75] Yang YL, Zhang W, Huang JD, Lin L, Lian HX, Lu YP, Wu JD, Wang SH. Purification and characterization of an extracellular xylanase from Aspergillus niger C3486. Afr J Microbiol Res, 2010, 4: 2249-2256.

[76] Zhang J, Viikari L. Impact of xylan on synergistic effects of xylanases and cellulases in enzymatic hydrolysis of lignocelluloses. Appl Biochem Biotechnol, 2014, 174: 1393-1402.

[77] Zhang J, Wang J, Fang C, Song F, Xin Y, Qu L, Ding K. Bacillus oceanisediminis sp. nov., isolated from marine sediment. Int J Syst Evol Microbiol, 2010, 60: 2924-2929.

[78] Zheng HC, Sun MZ, Meng LC, Pei HS, Zhang XQ, Yan Z, Sun JS. Purification and characterization of a thermostable xylanase from Paenibacillus sp. NF1 and its application in xylooligosaccharides production. J Microbiol Biotechnol, 2014, 24: 489-496.

[79] Zouari Ayadi D, Hmida Sayari A, Ben Hlima H, Ben Mabrouk S, Mezghani M, Bejar S. Improvement of Trichoderma reesei xylanase II thermal stability by serine to threonine surface mutations. Int J Biol Macromol, 2015, 72: 163-170.