Abstract
Facing the critical issue of high production costs for cellulase, numerous studies have focused on improving the efficiency of cellulase production by potential cellulolytic microorganisms using agricultural wastes as substrates, extremophilic cellulases, in particular, are crucial in the biorefinery process because they can maintain activity under harsh environmental conditions. This study aims to investigate the ability of a potential carboxymethylcellulose-hydrolyzing bacterial strain H1, isolated from an Algerian saline soil and identified as Bacillus velezensis, to use untreated olive mill wastes as a substrate for the production of an endo-1,4-β-glucanase. The enzyme was purified 44.9 fold using only two steps: ultrafiltration concentration and ion exchange chromatography, with final recovery of 80%. Its molecular mass was estimated to be 26 kDa by SDS-PAGE. Enzyme identification by LC–MS analysis showed 40% identity with an endo-1,3-1,4-β-glucanase of GH-16 family. The highest enzymatic activity was significantly measured on barley β-glucan (604.5 U/mL) followed by lichenan and carboxymethylcellulose as substrates, confirming that the studied enzyme is an endo-1,4-β-glucanase. Optimal enzymatic activity was at pH 6.0–6.5 and at 60–65 °C. It was fairly thermotolerant, retaining 76.9% of the activity at 70 °C, and halotolerant, retaining 70% of its activity in the presence of 4 M NaCl. The enzyme had a Vmax of 625 U/min/mL and a high affinity with barley β-glucan resulting a Km of 0.69 mg/mL. It also showed a significant ability to release cello-oligosaccharides. Based on such data, the H1 endo-1,4-β-glucanase may have significant commercial values for industry, argo-waste treatment, and other biotechnological applications.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Data availability
The gene sequence data of Bacillus velezensis strain H1 were deposited in the NCBI GenBank database and assigned the accession number OM510337 (16S RNA) and OM523097 (gyrA). The datasets generated during and/or analyzed during the current study are available from the corresponding author on reasonable request.
References
Afzal M, Qureshi MZ, Khan S, Khan MI, Ikram H, Ashraf A, Iqbal A, Qureshi NA (2019) Production, purification and optimization of cellulase by Bacillus licheniformis HI-08 isolated from the hindgut of wood-feeding termite. Int J Agric Biol 21:125–134
Ahmed SF, Mofijur M, Chowdhury SN, Nahrin M, Rafa N, Chowdhury AT, Nuzhat S, Ong HC (2022) Pathways of lignocellulosic biomass deconstruction for biofuel and value-added products production. Fuel 318:123618. https://doi.org/10.1016/j.fuel.2022.123618
Akita M, Kayatama K, Hatada Y, Ito S, Horikoshi K (2005) A novel β-glucanase gene from Bacillus halodurans C-125. FEMS Microbiol Lett 248:9–15. https://doi.org/10.1016/j.femsle.2005.05.009
Anwar Z, Gulfraz M, Irshad M (2014) Agro-industrial lignocellulosic biomass a key to unlock the future bio-energy: a brief review. J Radiat Res Appl Sci 7:163–173. https://doi.org/10.1016/j.jrras.2014.02.003
Bayer EA, Lamed R, Himmel ME (2007) The potential of cellulases and cellulosomes for cellulosic waste management. Curr Opin Biotechnol 18:237–245. https://doi.org/10.1016/j.copbio.2007.04.004
Bhayani JA, Ballicora MA (2022) Determination of dissociation constants of protein ligands by thermal shift assay. Biochem Biophys Res Commun 590:1–6. https://doi.org/10.1016/j.bbrc.2021.12.041
Boyce A, Walsh G (2007) Production, purification and application-relevant characterisation of an endo-1,3(4)-β-glucanase from Rhizomucor miehei. Appl Microbiol Biotechnol 76:835–841. https://doi.org/10.1007/s00253-007-1058-x
Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254. https://doi.org/10.1006/abio.1976.9999
Buratti C, Mousavi S, Barbanera M, Lascaro E, Cotana F, Bufacchi M (2016) Thermal behaviour and kinetic study of the olive oil production chain residues and their mixtures during co-combustion. Bioresour Technol 214:266–275. https://doi.org/10.1016/j.biortech.2016.04.097
Coker JA (2016) Extremophiles and biotechnology: current uses and prospects. F1000Res 5:396
Dar MA, Pawar KD, Rajput BP, Rahi P, Pandit RS (2019) Purification of a cellulase from cellulolytic gut bacterium, Bacillus tequilensis G9 and its evaluation for valorization of agro-wastes into added value byproducts. Biocatal Agric Biotechnol 20:101219. https://doi.org/10.1016/j.bcab.2019.101219
Dos Santos YQ, De Veras BO, De Franca AFJ, Gorlach-Lira K, Velasques J, Migliolo L, Dos Santos EA (2018) A new salt-tolerant thermostable cellulase from a marine Bacillus sp. strain. J Microbiol Biotechnol 28:1078–1085. https://doi.org/10.4014/jmb.1802.02037
Dotsenko G, Lange L (2017) Enzyme enhanced protein recovery from green biomass pulp. Waste Biomass Valori 8:1257–1264. https://doi.org/10.1007/s12649-016-9718-7
Fox GE, Wisotzkey JD, Jurtshuk P Jr (1992) How close is close: 16S rRNA sequence identity may not be sufficient to guarantee species identity. Int J Syst Evol Microbiol 42:166–170. https://doi.org/10.1099/00207713-42-1-166
Gabani P, Copeland E, Chandel AK, Singh OV (2012) Ultraviolet-radiation-resistant isolates revealed cellulose-degrading species of Cellulosimicrobium cellulans (UVP 1) and Bacillus pumilus (UVP 4). Biotechnol App Biochem 59:395–404. https://doi.org/10.1002/bab.1038
García GB, de Hoces MC, García CM, Palomino MTC, Gálvez AR, Martín-Lara MÁ (2014) Characterization and modeling of pyrolysis of the two-phase olive mill solid waste. Fuel Process Technol 126:104–111. https://doi.org/10.1016/j.fuproc.2014.04.020
Gares M, Hiligsmann S, Kacem Chaouche N (2020) Lignocellulosic biomass and industrial bioprocesses for the production of second generation bio-ethanol, does it have a future in Algeria? SN App Sci 2:1–19. https://doi.org/10.1007/s42452-020-03442-2
Garg R, Srivastava R, Brahma V, Verma L, Karthikeyan S, Sahni G (2016) Biochemical and structural characterization of a novel halotolerant cellulase from soil metagenome. Sci Rep 6:39634. https://doi.org/10.1038/srep39634
Gaur R, Tiwari S (2015) Isolation, production, purification and characterization of an organic-solvent-thermostable alkalophilic cellulase from Bacillus vallismortis RG-07. BMC Biotechnol 15:1–12. https://doi.org/10.1186/s12896-015-0129-9
Ghio S, Bradanini MB, Garrido MM, Ontañon OM, Piccinni FE, de Villegas RMD, Talia PM, Campos E (2020) Synergic activity of Cel8Pa β-1,4 endoglucanase and Bg1Pa β-glucosidase from Paenibacillus xylanivorans A59 in beta-glucan conversion. Biotechnol Rep 28:e00526. https://doi.org/10.1016/j.btre.2020.e00526
Gu X, Lu H, Zhang L, Meng X (2021) A Thermophilic GH5 endoglucanase from Aspergillus fumigatus and its synergistic hydrolysis of mannan-containing polysaccharides. Catalysts 11:862. https://doi.org/10.3390/catal11070862
Gupta M, Sharma M, Singh S, Gupta P, Bajaj BK (2015) Enhanced production of cellulase from Bacillus licheniformis K-3 with potential for saccharification of rice straw. Energy Technol 3:216–224. https://doi.org/10.1002/ente.201402137
Hakamada Y, Endo K, Takizawa S, Kobayashi T, Shirai T, Yamane T, Ito S (2002) Enzymatic properties, crystallization, and deduced amino acid sequence of an alkaline endoglucanase from Bacillus circulans. Biochim Biophys Acta Gen Subj 1570:174–180. https://doi.org/10.1016/S0304-4165(02)00194-0
Hrmova M, Fincher GB (2001) Structure-function relationships of β-D-glucan endo-and exohydrolases from higher plants. Plant Mol Biol 47:73–91. https://doi.org/10.1023/A:1010619128894
Huang Z, Ni G, Zhao X, Wang F, Qu M (2021) Characterization of a GH8 β-1,4-Glucanase from Bacillus subtilis B111 and Its Saccharification Potential for Agricultural Straws. J Microbiol Biotechnol 31:1446–1454. https://doi.org/10.4014/jmb.2105.05026
Ibrahim SNMM, Amalia SE, Agatha B, Lestari MY, Saputra AD, Ikmala NLF, Ryanto FHA, Dewi PA, Anam MK, Nafidiastri FA, Trikurniadewi N, Kumalasari DP, Tri E, Hariyanto G, Suwarni SA, Ni’matuzahroh, (2018) Screening and identifying of cellulolytic bacteria from Alas Purwo National Park. In AIP Conf Proc 2002:020064. https://doi.org/10.1063/1.5050160
Ibrahim AM, Hamouda RA, El-Naggar NEA, Al-Shakankery FM (2021) Bioprocess development for enhanced endoglucanase production by newly isolated bacteria, purification, characterization and in-vitro efficacy as anti-biofilm of Pseudomonas aeruginosa. Sci Rep 11:1–24. https://doi.org/10.1038/s41598-021-87901-9
Iqbal KMJ, Akhtar N, Khan MO, Khan MI (2022) Mix-method modelling of actors’ capacity for environmental sustainability and climate compatible development in energy sector. Environ Sci Pollut Res. https://doi.org/10.1007/s11356-022-19399-1
Kang SW, Park YS, Lee JS, Hong SI, Kim SW (2004) Production of cellulases and hemicellulases by Aspergillus niger KK2 from lignocellulosic biomass. Bioresour Technol 91:153–156. https://doi.org/10.1016/s0960-8524(03)00172-x
Kausar H, Ismail MR, Saud HM, Habib SH, Othman R, Mia G (2016) A novel lignocellulolytic bacterium for bioconversion of rice straw. Pak J Agric Sci 53:523–533
Khalid A, Ye M, Wei C, Dai B, Yang R, Huang S, Wang Z (2021) Production of β-glucanase and protease from Bacillus velezensis strain isolated from the manure of piglets. Prep Biochem Biotechnol 51:497–510. https://doi.org/10.1080/10826068.2020.1833344
Khandeparker R, Numan MT (2008) Bifunctional xylanases and their potential use in biotechnology. J Ind Microbiol Biotechnol 35:635–644. https://doi.org/10.1007/s10295-008-0342-9
Kim BK, Kim HJ, Lee JW (2013) Rapid statistical optimization of cultural conditions for mass production of carboxymethylcellulase by a newly isolated marine bacterium, Bacillus velezensis A-68 from rice hulls. J Life Sci 23:757–769. https://doi.org/10.5352/JLS.2013.23.6.757
Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685. https://doi.org/10.1038/227680a0
Lee YJ, Kim BK, Lee BH, Jo KI, Lee NK, Chung CH, Lee YC, Lee JW (2008) Purification and characterization of cellulase produced by Bacillus amyoliquefaciens DL-3 utilizing rice hull. Bioresour Technol 99:378–386. https://doi.org/10.1016/j.biortech.2006.12.013
Lineweaver H, Burk D (1934) The determination of enzyme dissociation constants. J Am Chem Soc 56:658–666. https://doi.org/10.1021/ja01318a036
Liu X, Ren B, Chen M, Wang H, Kokare CR, Zhou X, Wang J, Dai H, Song F, Liu M, Wang J, Wang S, Zhang L (2010) Production and characterization of a group of bioemulsifiers from the marine Bacillus velezensis strain H3. App Microbiol Biotechnol 87:1881–1893. https://doi.org/10.1007/s00253-010-2653-9
Liu D, Zhang R, Yang X, Xu Y, Tang Z, Tian W, Shen Q (2011) Expression, purification and characterization of two thermostable endoglucanases cloned from a lignocellulosic decomposing fungi Aspergillus fumigatus Z5 isolated from compost. Protein Expr Purif 79:176–186. https://doi.org/10.1016/j.pep.2011.06.008
Liu Y, Guo H, Wu Y, Qin W (2018) Purification and characterizations of a novel recombinant Bacillus velezensis endoglucanase by aqueous two-phase system. Bioresour Bioprocess 5:1–11. https://doi.org/10.1186/s40643-018-0204-x
Liu Y, Guo H, Gu J, Qin W (2019) Optimize purification of a cellulase from Bacillus velezensis A4 by aqueous two-phase system (ATPS) using response surface methodology. Process Biochem 87:196–203. https://doi.org/10.1016/j.procbio.2019.08.017
McCleary BV, McGeough P (2015) A comparison of polysaccharide substrates and reducing sugar methods for the measurement of endo-1,4-β-xylanase. App Biochem Biotechnol 177:1152–1163. https://doi.org/10.1007/s12010-015-1803-z
Miller GL (1959) Use of dinitrosalicylic acid reagent for determination of reducing sugar. Anal Chem 31:426–428
Miotto LS, de Rezende CA, Bernardes A, Serpa VI, Tsang A, Polikarpov I (2014) The characterization of the endoglucanase Cel12A from Gloeophyllum trabeum reveals an enzyme highly active on β-glucan. PLoS ONE 9:e108393. https://doi.org/10.1371/journal.pone.0108393
Nair AS, Al-Battashi H, Al-Akzawi A, Annamalai N, Gujarathi A, Al-Bahry S, Dhillon GS, Nallusamy S (2018) Waste office paper: a potential feedstock for cellulase production by a novel strain Bacillus velezensis ASN1. Waste Manag 79:491–500. https://doi.org/10.1016/j.wasman.2018.08.014
Pérez J, Munoz-Dorado J, De la Rubia TDLR, Martinez J (2002) Biodegradation and biological treatments of cellulose, hemicellulose and lignin: an overview. Int Microbiol 5:53–63. https://doi.org/10.1007/s10123-002-0062-3
Phachan N, Fiala K, Apirakakorn J (2017) Isolation of cellulolytic clostridia and their performance for one-step butanol production from sugarcane bagasse. Energy Procedia 138:163–168. https://doi.org/10.1016/j.egypro.2017.10.144
Pham VHT, Kim J, Shim J, Chang S, Chung W (2022) Coconut mesocarp-based lignocellulosic waste as a substrate for cellulase production from high promising multienzyme-producing Bacillus amyloliquefaciens FW2 without pretreatments. Microorganisms 10:327. https://doi.org/10.3390/microorganisms10020327
Planas A (2000) Bacterial 1,3–1,4-β-glucanases: structure, function and protein engineering. Biochim Biophys Acta Prot Struct Mol Enzym 1543:361–382. https://doi.org/10.1016/s0167-4838(00)00231-4
Plecha S, Hall D, Tiquia-Arashiro SM (2013) Screening for novel bacteria from the bioenergy feedstock switchgrass (Panicum virgatum L.). Environ Technol 34:1895–1904. https://doi.org/10.1080/09593330.2013.818701
Pramanik SK, Mahmud S, Paul GK, Jabin T, Naher K, Salah Uddin Md, Zaman S, Abu Saleh MA (2021) Fermentation optimization of cellulase production from sugarcane bagasse by Bacillus pseudomycoides and molecular modeling study of cellulase. Curr Res Microb Sci 2:100013. https://doi.org/10.1016/j.crmicr.2020.100013
Rahman SS, Siddique R, Tabassum N (2017) Isolation and identification of halotolerant soil bacteria from coastal Patenga area. BMC Res Notes 10:1–6. https://doi.org/10.1186/s13104-017-2855-7
Rastogi RP, Pandey A, Larroche C, Madamwar D (2018) Algal green energy—R&D and technological perspectives for biodiesel production. Renew Sustain Energy Rev 82:2946–2969. https://doi.org/10.1016/j.rser.2017.10.038
Rathnan RK, John D (2021) Isolation, screening, identification and optimized production of extracellular cellulase from Bacillus subtilis using cellulosic waste as carbon source. J Microbiol Biotechnol Food Sci 2021:2383–2386
Rattu G, Joshi S, Satyanarayana T (2016) Bifunctional recombinant cellulase–xylanase (rBhcell-xyl) from the polyextremophilic bacterium Bacillus halodurans TSLV1 and its utility in valorization of renewable agro-residues. Extremophiles 20:831–842. https://doi.org/10.1007/s00792-016-0870-6
Rodríguez G, Lama A, Rodríguez R, Jiménez A, Guillén R, Fernández-Bolaños J (2008) Olive stone an attractive source of bioactive and valuable compounds. Bioresour Technol 99:5261–5269. https://doi.org/10.1016/j.biortech.2007.11.027
Ruiz-Garcia C, Bejar V, Martinez-Checa F, Llamas I, Quesada E (2005) Bacillus velezensis sp. nov., a surfactant-producing bacterium isolated from the river Velez in Malaga, southern Spain. Int J Syst Evol Microbiol 55:191–195. https://doi.org/10.1099/ijs.0.63310-0
Sadhu S, Saha P, Sen SK, Mayilraj S, Maiti TK (2013) Production, purification and characterization of a novel thermotolerant endoglucanase (CMCase) from Bacillus strain isolated from cow dung. Springerplus 2:1–10. https://doi.org/10.1186/2193-1801-2-10
Saritha M, Arora A, Lata, (2012) Biological pretreatment of lignocellulosic substrates for enhanced delignification and enzymatic digestibility. Indian J Microbiol 52:122–130. https://doi.org/10.1007/s12088-011-0199-x
Schimming S, Schwarz WH, Staudenbauer WL (1991) Properties of a thermoactive β-1,3–1,4-glucanase (lichenase) from Clostridium thermocellum expressed in Escherichia coli. Biochem Biophys Res Commun 177:447–452. https://doi.org/10.1016/0006-291x(91)92004-4
Sharma M, Bajaj BK (2014) Cellulase production from Bacillus subtilis MS 54 and its potential for saccharification of biphasic-acid-pretreated rice straw. J Biobased Mater Bioenergy 8:449–456. https://doi.org/10.1166/jbmb.2014.1458
Silva TP, Fabiana S, Dos Santos CWV, Franco M, Caetano LC, Pereira HJV (2018) Production, purification, characterization and application of a new halotolerant and thermostable endoglucanase of Botrytis ricini URM 5627. Bioresour Technol 270:263–269. https://doi.org/10.1016/j.biortech.2018.09.022
Singh S, Moholkar VS, Goyal A (2013) Isolation, identification, and characterization of a cellulolytic Bacillus amyloliquefaciens strain SS35 from rhinoceros dung. Int Sch Res Notices. https://doi.org/10.1155/2013/728134
Sulyman AO, Igunnu A, Malomo SO (2020) Isolation, purification and characterization of cellulase produced by Aspergillus niger cultured on Arachis hypogaea shells. Heliyon 6:e05668. https://doi.org/10.1016/j.heliyon.2020.e05668
Sun Y, Cheng J (2002) Hydrolysis of lignocellulosic materials for ethanol production: a review. Bioresour Technol 83:1–11. https://doi.org/10.1016/S0960-8524(01)00212-7
Sun J, Wang H, Lv W, Ma C, Lou Z, Yao H, Dai Y (2012) Cloning and expression of a thermostable β-1,3–1,4-glucanase from Bacillus amyloliquefaciens ATCC 23350. Ann Microbiol 62:1235–1242. https://doi.org/10.1007/s13213-011-0366-7
Teather RM, Wood PJ (1982) Use of Congo red-polysaccharide interactions in enumeration and characterization of cellulolytic bacteria from the bovine rumen. App Environ Microbiol 43:777–780. https://doi.org/10.1128/aem.43.4.777-780.1982
Tesfaye B, Tefera T (2017) Extraction of essential oil from neem seed by using soxhlet extraction methods. Int J Adv Engi Manag Sci 3:239870
Thomas L, Ram H, Singh VP (2018) Inducible cellulase production from an organic solvent tolerant Bacillus sp. SV1 and evolutionary divergence of endoglucanase in different species of the genus Bacillus. Braz J Microbiol 49:429–442. https://doi.org/10.1016/j.bjm.2017.05.010
Trivedi N, Gupta V, Kumar M, Kumari P, Reddy CRK, Jha B (2011) An alkali-halotolerant cellulase from Bacillus flexus isolated from green seaweed Ulva lactuca. Carbohydr Polym 83:891–897. https://doi.org/10.1016/j.carbpol.2010.08.069
Van Soest PJ (1994) Nutritional ecology of the ruminant. Cornell University Press. https://doi.org/10.7591/9781501732355
Volpe R, Messineo A, Millan M, Volpe M, Kandiyoti R (2015) Assessment of olive wastes as energy source: pyrolysis, torrefaction and the key role of H loss in thermal breakdown. Energy 82:119–127. https://doi.org/10.1016/j.energy.2015.01.011
Wang JL, Ruan H, Zhang HF, Zhang Q, Zhang HB, He GQ, Shen SR (2007) Characterization of a thermostable and acidic-tolerable β-glucanase from aerobic fungi Trichoderma koningii ZJU-T. J Food Sci 72:C452–C456. https://doi.org/10.1111/j.1750-3841.2007.00549.x
Xu J, He B, Wu B, Wang B, Wang C, Hu L (2014) An ionic liquid tolerant cellulase derived from chemically polluted microhabitats and its application in in situ saccharification of rice straw. Bioresour Technol 157:166–173. https://doi.org/10.1016/j.biortech.2014.01.102
Xu T, Zhu T, Li S (2016) β-1,3–1,4-glucanase gene from Bacillus velezensis ZJ20 exerts antifungal effect on plant pathogenic fungi. World J Microbiol Biotechnol 32:1–9. https://doi.org/10.1007/s11274-015-1985-0
Yan J, Liu W, Li Y, Lai HL, Zheng Y, Huang JW, Chen CC, Chen Y, Jin J, Li H, Guo RT (2016) Functional and structural analysis of Pichia pastoris-expressed Aspergillus niger 1,4-β-endoglucanase. Biochem Biophys Res Commun 475:8–12. https://doi.org/10.1016/j.bbrc.2016.05.012
Ye M, Sun L, Yang R, Wang Z, Qi K (2017) The optimization of fermentation conditions for producing cellulase of Bacillus amyloliquefaciens and its application to goose feed. R Soc Open Sci 4:171012. https://doi.org/10.1098/rsos.171012
Yoo DH, Lee BH, Chang PS, Lee HG, Yoo SH (2007) Improved quantitative analysis of oligosaccharides from lichenase-hydrolyzed water-soluble barley β-glucans by high-performance anion-exchange chromatography. J Agric Food Chem 55:1656–1662. https://doi.org/10.1021/jf062603l
Zarei O, Dastmalchi S, Hamzeh-Mivehroud M (2016) A simple and rapid protocol for producing yeast extract from Saccharomyces cerevisiae suitable for preparing bacterial culture media. Iran J Pharm Res 15:907
Zhang M, Chen XL, Zhang ZH, Sun CY, Chen LL, He HL, Zhou BC, Zhang YZ (2009) Purification and functional characterization of endo-β-mannanase MAN5 and its application in oligosaccharide production from konjac flour. Appl Microbiol Biotechnol 83:865–873. https://doi.org/10.1007/s00253-009-1920-0
Zhao K, Guo LZ, Lu WD (2012) Extracellular production of novel halotolerant, thermostable, and alkali-stable carboxymethyl cellulase by marine bacterium Marinimicrobium sp. LS-A18. App Biochem Biotechnol 168:550–567. https://doi.org/10.1007/s12010-012-9796-3
Acknowledgements
We Acknowledge Dr. Roumaissa Zenzen for helping us to collect the olive mill wastes.
Funding
This work was supported by the Ministry of Higher Education and Scientific Research of Algeria, and Wallonie-Bruxelles International (WBI).
Author information
Authors and Affiliations
Contributions
HD: investigation, data curation, methodology, writing—original draft, writing—review and editing. SF: project administration, supervisor, investigation, methodology, validation, formal analysis, writing—review and editing. CVW: investigation, methodology, validation, formal analysis, writing—review and editing. YO: data curation, formal analysis, and methodology. SH: project administration, formal analysis, and validation. BC: formal analysis and software. RC: writing—review and editing, validation. mg: project contribution. NKC: project administration, supervisor, investigation, methodology, validation, formal analysis, writing—review and editing.
Corresponding author
Ethics declarations
Conflict of interest
The authors declare that they have no competing interests.
Additional information
Communicated by Erko Stackebrandt.
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.
About this article
Cite this article
Djelid, H., Flahaut, S., Vander Wauven, C. et al. Production of a halotolerant endo-1,4-β-glucanase by a newly isolated Bacillus velezensis H1 on olive mill wastes without pretreatment: purification and characterization of the enzyme. Arch Microbiol 204, 681 (2022). https://doi.org/10.1007/s00203-022-03300-2
Received:
Revised:
Accepted:
Published:
DOI: https://doi.org/10.1007/s00203-022-03300-2