Abstract
In this report, protease-producing haloalkaliphilic bacteria from sea water have been investigated. Seven bacterial strains belonging to Geomicrobium halophilum, Oceanobacillus oncorhynchi, and Oceanobacillus khimchii were isolated and characterized based on their colony characteristics, cell morphology, biochemical properties, antibiotic sensitivity, and 16S rRNA gene sequence analysis. The analysis based on 53 phenotypic characters (cluster analysis) generated a phenogram establishing correlation with the phylogenetic placement. A maximal level of phenotypic similarity among the protease producing bacteria was evident at only 46% in the phenogram. An overall coherence was evident between the patterns reflected in phenogram and phylogram. Four isolates of different geographical locations belonging to Geomicrobium halophilum were scattered in the phenogram, displaying significant variations in their phenotypes. Further, stability and catalysis of the extracellular alkaline proteases of these isolates were studied. The bacteria were able to grow in the range of pH 7-10 and temperatures between 37-55 °C. The proteases were active and stable at pH 7-13 with the optima between 9.5 and 10.5, although a rapid loss of activity was observed at pH 6.5 and below. Most of the proteases had temperature optima at around 60 °C, while they were stable in the range of 37-60 °C, with the retention of 100-80% activities after 1h of incubation at 60-90 °C. As a unique feature, the enzymes were resistant to urea denaturation (2-8 M) for a period of 2-24 h. The addition of NaCl had profound effect on the resistance against chemical denaturation.
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References
Amoozegar MA, Siroosi M, Atashgahi S, Smidt H, Ventosa A (2017) Systematics of haloarchaea and biotechnological potential of their hydrolytic enzymes. Microbiology 63:623–645. https://doi.org/10.1099/mic.0.000463
Arahal DR, Dewhirst FE, Paster BJ, Volcani BE, Ventosa A (1996) Phylogenetic analyses of some extremely halophilic archaea isolated from Dead Sea water, determined on the basis of their 16S rRNA sequences. Appl Environ Microbiol 62:3779–3786. https://doi.org/10.1128/aem.62.10.3779-3786.1996
Barzkar N (2020) Marine microbial alkaline protease: an efficient and essential tool for various industrial applications. Int J Biol Macromol 161:1216–1229. https://doi.org/10.1016/j.ijbiomac.2020.06.072
Baweja M, Singh PK, Sadaf A, Tiwari R, Nain L, Khare SK, Shukla P (2017) Cost effective characterization process and molecular dynamic simulation of detergent compatible alkaline protease from Bacillus pumilus strain MP27. Process Biochem 58:199–203. https://doi.org/10.1016/j.procbio.2017.04.024
Bhatt HB, Begum AM, Chintalapati S, Chintalapati VR, Singh SP (2017) Desertibacillus haloalkaliphilus gen. nov. sp. nov., isolated from a saline desert. Int J Syst Evol Microbiol 67:4435–4442. https://doi.org/10.1099/ijsem.0.002310
Bhatt HB, Gohel SD, Singh SP (2018) Phylogeny, novel bacterial lineage and enzymatic potential of haloalkaliphilic bacteria from the saline coastal desert of Little Rann of Kutch, Gujarat, India. 3 Biotech 8: 53. https://doi.org/10.1007/s13205-017-1075-0
Bhatt HB, Singh SP (2016) Phylogenetic and phenogram based diversity of haloalkaliphilic bacteria from the saline desert. In: Bhukya B, Tangutur AD (eds) Microbial biotechnology-technological challenges and developmental trends. Apple Academic Press, Oakville, pp 373–386. 10.1201/b19978-24
Bhatt HB, Singh SP (2020) Cloning, expression, and structural elucidation of a biotechnologically potential alkaline serine protease from a newly isolated haloalkaliphilic Bacillus lehensis JO-26. Front Microbiol 11:941. https://doi.org/10.3389/fmicb.2020.00941
Bradford MM (1976) A rapid and sensitive method for quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254. https://doi.org/10.1016/0003-2697(76)90527-3
Briki S, Hamdi O, Landoulsi A (2016) Enzymatic dehairing of goat skins using alkaline protease from Bacillus sp. SB12. Protein Expr Purif 121:9–16. https://doi.org/10.1016/j.pep.2015.12.021
Cappuccino J, Sherman N (2004) Microbiology: A Laboratory Manual. 7th edn Benjamin Cummings Science publishing, California.
Cheng Q, Xu F, Hu N, Liu X, Liu Z (2015) A novel Ca2+-dependent alkaline serine-protease (Bvsp) from Bacillus sp. with high fibrinolytic activity. J Mol Catal B Enzym 117:69–74. https://doi.org/10.1016/j.molcatb.2015.04.006
Contesini FJ, Melo RR, Sato HH (2018) An overview of Bacillus proteases: from production to application. Crit Rev Biotechnol 38:321–334. https://doi.org/10.1080/07388551.2017.1354354
Dalmaso GZ, Ferreira D, Vermelho AB (2015) Marine extremophiles: a source of hydrolases for biotechnological applications. Mar Drugs 13:1925–1965. https://doi.org/10.3390/md13041925
Demergasso C, Casamayor EO, Chong G, Galleguillos P, Escudero L, Pedrós-Alió C (2004) Distribution of prokaryotic genetic diversity in athalassohaline lakes of the Atacama Desert, Northern Chile. FEMS Microbiol Ecol 48:57–69. https://doi.org/10.1016/j.femsec.2003.12.013
Dodia MS, Bhimani HG, Rawal CM, Joshi RH, Singh SP (2008a) Salt dependent resistance against chemical denaturation of alkaline protease from newly isolated haloalkaliphilic Bacillus sp. Bioresour Technol 99:6223–6227. https://doi.org/10.1016/j.biortech.2007.12.020
Dodia MS, Joshi RH, Patel RK, Singh SP (2006) Characterization and stability of extracellular alkaline proteases from moderately halophilic and alkaliphilic Bacteria isolated from saline habitat of coastal Gujarat, India. Braz J Microbiol 37:276–282. https://doi.org/10.1590/S1517-83822006000300015
Dodia MS, Rawal CM, Bhimani HG, Joshi RH, Khare SK, Singh SP (2008b) Purification and stability characteristics of an alkaline serine protease from a newly isolated Haloalkaliphilic bacterium sp. AH-6. J Ind Microbiol Biotechnol 35:121–131. https://doi.org/10.1007/s10295-007-0273-x
Dorra G, Ines K, Imen BS, Laurent C, Sana A, Olfa T, Pascal C, Thierry J, Ferid L (2018) Purification and characterization of a novel high molecular weight alkaline protease produced by an endophytic Bacillus halotolerans strain CT2. Int J Biol Macromol 111:342–351. https://doi.org/10.1016/j.ijbiomac.2018.01.024
Duckworth AW, Grant WD, Jones BE, Steenbergen RV (1996) Phylogenetic diversity of Soda Lake alkaliphiles. FEMS Microbiol Ecol 19:181–191. https://doi.org/10.1111/j.1574-6941.1996.tb00211.x
Dumorné K, Córdova DC, Astorga-Eló M, Renganathan P (2017) Extremozymes: a potential source for industrial applications. J Microbiol Biotechnol 27(4):649–659. https://doi.org/10.4014/jmb.1611.11006
Dwivedi P, Sharma AK, Singh SP (2021) Biochemical properties and repression studies of an alkaline serine protease from a haloalkaliphilic actinomycete, Nocarpdiopsis dassonvillei subsp. albirubida OK-14. Biocatal Agric Biotechnol 9:102059. https://doi.org/10.1016/j.bcab.2021.102059
Echigo A, Minegishi H, Mizuki T, Kamekura M, Usami R (2010) Geomicrobium halophilum gen. nov., sp. nov., a moderately halophilic and alkaliphilic bacterium isolated from soil. Int J Syst Evol Microbiol. 60(4):990-5. https://doi.org/10.1099/ijs.0.63483-0
Foti MJ, Sorokin DY, Zacharova EE, Pimenov NV, Kuenen JG, Muyzer G (2008) Bacterial diversity and activity along a salinity gradient in soda lakes of the Kulunda steppe (Altai, Russia). Extremophiles 12:133–145. https://doi.org/10.1007/s00792-007-0117-7
Gimenez MI, Studdert CA, Sanchez J, De Castro RE (2000) Extracellular protease of Natrialba magadii: purification and biochemical characterization. Extremophiles 4:181–188. https://doi.org/10.1007/s007920070033
Gohel SD, Singh SP (2018) Thermodynamics of a Ca2+ dependent, highly thermostable and detergent compatible purified alkaline serine protease from Nocardiopsis xinjiangensis strain OM-6. Int J Biol Macromol 113:565–574. https://doi.org/10.1016/j.ijbiomac.2018.02.157
Goris J, Konstantinidis KT, Klappenbach JA, Coenye T, Vandamme P, Tiedje JM (2007) DNA–DNA hybridization values and their relationship to whole-genome sequence similarities. Int J Syst Evol Microbiol 57(1):81–91. https://doi.org/10.1099/ijs.0.64483-0
Haddar A, Agrebi R, Bougatef A, Hmidet N, Sellami-Kamoun A, Nasri M (2009) Two detergent stable alkaline serine-proteases from Bacillus mojavensis A21: purification, characterization and potential application as a laundry detergent additive. Bioresour Technol 100:3366–3373. https://doi.org/10.1016/j.biortech.2009.01.061
Hagihara B (1958) The Enzymes Vol. 4. Academic Press Inc., New York.
Haki GD, Rakshit SK (2003) Developments in industrially important thermostable enzymes. Bioresour Technol 89:17–34. https://doi.org/10.1016/S0960-8524(03)00033-6
Hammer O, Harper DAT, Ryan PD (2001) PAST-Palaeontological statistics. www.uv.es/~pardomv/pe/2001_1/past/pastprog/past.pdf, acessado em, 25(07): 2009.
Hedi A, Sadfi N, Fardeau ML, Rebib H, Cayol JL, Ollivier B, Boudabous A (2009) Studies on the biodiversity of halophilic microorganisms isolated from El-Djerid Salt Lake (Tunisia) under aerobic conditions. Int J Microbiol. https://doi.org/10.1155/2009/731786
Hutcheon GW, Vasisht N, Bolhuis A (2005) Characterization of a highly stable α-amylase from the halophilic archaeon Haloarcula hispanica. Extremophiles 9(6):487–495. https://doi.org/10.1007/s00792-005-0471-2
Jaouadi B, Ellouz-Chaabouni S, Rhimi M, Bejar S (2008) Biochemical and molecular characterization of a detergent-stable serine alkaline protease from Bacillus pumilus CBS with high catalytic efficiency. Biochimie. 90(9):1291–1305. https://doi.org/10.1016/j.biochi.2008.03.004
Jellouli K, Ghorbel-Bellaaj O, Ayed HB, Manni L, Agrebi R, Nasri M (2011) Alkaline-protease from Bacillus licheniformis MP1: purification, characterization and potential application as a detergent additive and for shrimp waste deproteinization. Process Biochem 46(6):1248–1256. https://doi.org/10.1016/j.procbio.2011.02.012
Joshi S, Satyanarayana T (2013) Characteristics and applications of a recombinant alkaline serine protease from a novel bacterium Bacillus lehensis. Bioresour Technol 131:76–85. https://doi.org/10.1016/j.biortech.2012.12.124
Karan R, Khare SK (2010) Purification and characterization of a solvent-stable protease from Geomicrobium sp. EMB2. Environmental technology. 31(10):1061-72. https://doi.org/10.1080/09593331003674556
Karan R, Singh SP, Kapoor S, Khare SK (2011) A novel organic solvent tolerant protease from a newly isolated Geomicrobium sp. EMB2 (MTCC 10310): production optimization by response surface methodology. N Biotechnol 28(2):136–145. https://doi.org/10.1016/j.nbt.2010.10.007
Kennedy AC, Papendick RI (1995) Microbial characteristics of soil quality. J Soil Water Conserv 50(3):243–248
Kennedy J, Marchesi JR, Dobson AD (2008) Marine metagenomics: Strategies for the discovery of novel enzymes with biotechnological applications from marine environments. Microb Cell Fact 7(27). https://doi.org/10.1186/1475-2859-7-27
Kim TY, Kim SJ, Park SJ, Kim JG, Cha IT, Jung MY, Lee SA, Roh SW, Yim KJ, Itoh T, Rhee SK (2013) Natronomonas gomsonensis sp. nov. isolated from a solar saltern. Antonie Van Leeuwenhoek 104:627–635. https://doi.org/10.1007/s10482-013-9970-9
Kimura M (1980) A simple method for estimating evolutionary rates of base substitutions through comparative studies of nucleotide sequences. J Mol Evol 16:111–120. https://doi.org/10.1007/BF01731581
Lam KS (2006) Discovery of novel metabolites from marine actinomycetes. Curr Opin Microbiol 9:245–251. https://doi.org/10.1016/j.mib.2006.03.004
Littlechild JA (2015) Enzymes from extreme environments and their industrial applications. Front Bioeng Biotechnol 3:161. https://doi.org/10.3389/fbioe.2015.00161
Margesin R, Schinner F (2001) Potential of halotolerant and halophilic microorganisms for biotechnology. Extremophiles 5:73–83. https://doi.org/10.1007/s007920100184
Oren A (2002) Molecular ecology of extremely halophilic Archaea and Bacteria. FEMS Microbiol Ecol 39:1–7. https://doi.org/10.1111/j.1574-6941.2002.tb00900.x
Pandey S, Rakholiya KD, Raval VH, Singh SP (2012) Catalysis and stability of an alkaline protease from a haloalkaliphilic bacterium under non-aqueous conditions as a function of pH, salt and temperature. J Biosci Bioeng 114(3):251–256. https://doi.org/10.1016/j.jbiosc.2012.03.003
Pandey S, Sharma AK, Solanki KP, Singh SP (2018) Catalysis and stability of an extracellular α-amylase from a haloalkaliphilic bacterium as a function of the organic solvents at different pH, salt concentrations and temperatures. Indian J Geo-Mar Sci 47:240–248 http://nopr.niscair.res.in/handle/123456789/43431
Patel RK, Dodia MS, Joshi RH, Singh SP (2006b) Purification and characterization of alkaline protease from a newly isolated haloalkaliphilic Bacillus sp. Process Biochem 41:2002–2009. https://doi.org/10.1007/s12275-011-0427-4
Patel RK, Dodia MS, Joshi RH, Singh SP (2006a) Production of extracellular halo-alkaline protease from a newly isolated Haloalkaliphilic Bacillus sp. isolated from seawater in Western India. World J Microbiol Biotechnol 24:375–382. https://doi.org/10.1007/s11274-005-9044-x
Patel RK, Dodia MS, Singh SP (2005) Extracellular alkaline protease from a newly isolated haloalkaliphilic Bacillus sp.: Production and optimization. Process Biochem 40:3569–3575. https://doi.org/10.1016/j.procbio.2005.03.049
Phillips K, Zaidan F, Elizondo OR, Lowe KL (2012) Phenotypic characterization and 16S rDNA identification of culturable non-obligate halophilic bacterial communities from a hypersaline lake, La Sal del Rey, in extreme South Texas (USA). Aquat Biosyst 8:5. https://doi.org/10.1186/2046-9063-8-5
Polosina YY, Zamyatkin DF, Kostyukova AS, Filimonov VV, Fedorov OV (2002) Stability of Natrialba magadii NDP kinase: comparisons with other halophilic proteins. Extremophiles. 6(2):135–142. https://doi.org/10.1007/s007920100232
Purohit MK, Singh SP (2014) Cloning, over expression and functional attributes of serine proteases from Oceanobacillus iheyensis OM A18 and Haloalkaliphilic bacterium OM E12. Process Biochem 49(1):61–68. https://doi.org/10.1016/j.procbio.2013.07.009
Qeshmi FI, Homaei A, Fernandes P, Javadpour S (2018) Marine microbial L-asparaginase: biochemistry, molecular approaches and applications in tumor therapy and in food industry. Microbiol Res 208:99–112. https://doi.org/10.1016/j.micres.2018.01.011
Rathore DS, Sheikh MA, Gohel SD, Singh SP (2021) Genetic and Phenotypic Heterogeneity of the Nocardiopsis alba Strains of Seawater. Curr Microbiol 78(4):1377–1387. https://doi.org/10.1007/s00284-021-02420-0
Rathore DS, Singh SP (2021) Kinetics of growth and co-production of amylase and protease in novel marine actinomycete, Streptomyces lopnurensis KaM5. Folia Microbiol 66(3):303–316. https://doi.org/10.1007/s12223-020-00843-z
Raval VH, Bhatt HB, Singh SP (2018) Adaptation Strategies in Halophilic Bacteria. In Extremophiles (pp. 137-164). CRC Press.
Raval VH, Pillai S, Rawal CM, Singh SP (2014) Biochemical and structural characterization of a detergent-stable serine alkaline protease from seawater haloalkaliphilic bacteria. Process Biochem 49(6):955–962. https://doi.org/10.1016/j.procbio.2014.03.014
Raval VH, Rawal CM, Pandey S, Bhatt HB, Dahima BR, Singh SP (2015) Cloning, heterologous expression and structural characterization of an alkaline serine protease from sea water haloalkaliphilic bacterium. Ann Microbiol 65(1):371–381. https://doi.org/10.1007/s13213-014-0869-0
Romano I, Giordano A, Lama L, Nicolaus B, Gambacorta A (2005) Halomonas campaniensis sp. nov. a haloalkaliphilic bacterium isolated from a mineral pool of Campania Region, Italy. Syst Appl Microbiol 28:610–618. https://doi.org/10.1016/j.syapm.2005.03.010
Sambrook J, Russell DW (2001) Molecular Cloning: A Laboratory Manual, 3rd edn. Cold Spring Harbor Laboratory Press, New York
Setyorini E, Takenaka S, Murakami S, Aoki K (2006) Purification and characterization of two novel halotolerant extracellular proteases from Bacillus subtilis strain FP-133. Biosci Biotechnol Biochem 70(2):433–440. https://doi.org/10.1271/bbb.70.433
Sharma AK, Kikani BA, Singh SP (2020) Biochemical, thermodynamic and structural characteristics of a biotechnologically compatible alkaline protease from a haloalkaliphilic, Nocardiopsis dassonvillei OK-18. Int J Biol Macromol 153:680–696. https://doi.org/10.1016/j.ijbiomac.2020.03.006
Singh AK, Chhatpar HS (2011) Purification, characterization and thermodynamics of antifungal protease from Streptomyces sp. A6. J Basic Microbiol 51:424–432. https://doi.org/10.1002/jobm.201000310
Singh SP, Purohit MK, Raval VH, Pandey S, Akbari VG, Rawal CM (2010) Capturing the potential of Haloalkaliphilic bacteria from the saline habitats through culture dependent and metagenomic approaches. Applied Microbiology and Microbial Biotechnology; Méndez-Vilas, A., Ed. 2010:81-7.
Singh SP, Raval V, Purohit MK (2013) Strategies for the Salt Tolerance in Bacteria and Archeae and its Implications in Developing Crops for Adverse Conditions. In Plant Acclimation to Environmental Stress (pp. 85-99). Springer, New York, NY.
Singh SP, Raval VH, Purohit MK, Thumar JT, Gohel SD, Pandey S, ... & Rawal CM (2012) Haloalkaliphilic bacteria and actinobacteria from the saline habitats: new opportunities for biocatalysis and bioremediation. In Microorganisms in Environmental Management (pp. 415-429). Springer, Dordrecht.
Sinha R, Khare SK (2013) Characterization of detergent compatible protease of a halophilic Bacillus sp. EMB9: differential role of metal ions in stability and activity. Bioresour Technol 145:357–361. https://doi.org/10.1016/j.biortech.2012.11.024
Sinsuwan S, Rodtong S, Yongsawatdigul J (2010) Purification and characterization of a salt-activated and organic solvent-stable heterotrimer proteinase from Virgibacillus sp. SK33 isolated from thai fish sauce. J Agric Food Chem 58(1):248–256. https://doi.org/10.1021/jf902479k
Stackebrandt E, Ebers J (2006) Taxonomic parameters revisited: tarnished gold standards. Microbiol Today 33:152–155
Tavano OL, Berenguer-Murcia A, Secundo F, Fernandez-Lafuente R (2018) Biotechnological applications of proteases in food technology. Compr Rev Food Sci Food Saf 17(2):412–436. https://doi.org/10.1111/1541-4337.12326
Thakrar FJ, Singh SP (2019) Catalytic, thermodynamic and structural properties of an immobilized and highly thermostable alkaline protease from a haloalkaliphilic actinobacteria, Nocardiopsis alba TATA-5. Bioresour Technol 278:150–158. https://doi.org/10.1016/j.biortech.2019.01.058
Whon TW, Jung MJ, Roh SW, Nam YD, Park EJ, Shin KS, Bae JW (2010) Oceanobacillus kimchii sp. nov. isolated from a traditional Korean fermented food. J. Microbiol. 48(6):862-6. https://doi.org/10.1007/s12275-010-0214-7
Yumoto I, Hirota K, Nodasaka Y, Nakajima K (2005) Oceanobacillus oncorhynchi sp. nov., a halotolerant obligate alkaliphile isolated from the skin of a rainbow trout (Oncorhynchus mykiss), and emended description of the genus Oceanobacillus. Int J Syst Evol Microbiol. 55(4):1521-4. https://doi.org/10.1099/ijs.0.013268-0
Zeng R, Zhang R, Zhao J, Lin N (2003) Cold active serine alkaline protease from the Psychrophilic bacterium Pseudomonas strain DY-A: enzyme purification and characterization. Extremophiles 7:335–337. https://doi.org/10.1007/s00792-003-0323-x
Zhang C, Kim SK (2010) Research and application of marine microbial enzymes: status and prospects. Mar Drugs 8(6):1920–1934. https://doi.org/10.3390/md8061920
Zhou C, Qin H, Chen X, Zhang Y, Xue Y, Ma Y (2018) A novel alkaline protease from alkaliphilic Idiomarina sp. C9-1 with potential application for eco-friendly enzymatic dehairing in the leather industry. Sci Rep 8(1):1–8. https://doi.org/10.1038/s41598-018-34416-5
Acknowledgements
VR is thankful to Scientific and Engineering Research Board (SERB) (YSS/2015/001255), New Delhi, India for the Young Scientist Project. HB is thankful to the Council of Scientific and Industrial Research (CSIR), New Delhi, India for the award of Direct Senior Research Fellowship (CSIR–SRF) and the award of the UGC-Meritorious Fellowship. We acknowledge the support under the UGC-Centre of Advanced Study Program (No.F.5-4/2012(SAP-II), DBT Multi-Institutional Project, DST-FIST program (N0.SR/FST/LSI-281/2006) and MoES Net-Working Project of the Govt. of India. SPS acknowledges International Travel Supports from SERB-DST, UGC, CSIR and DBT to present his research in Hamburg (Germany), Brisbane (Australia), Cape town (South Africa), and Kyoto (Japan). SPS further acknowledges the award of the UGC-BSR Faculty Fellowship. Financial and intracerebral support from the Saurashtra university is also duly acknowledged.
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All authors contributed to study conception and design. RHJ performed experiments related to enzyme stability and catalysis. HBB contributed to analysis of numerical taxonomy and phylogeny. RHJ and VHR compiled and interpreted the data and wrote the manuscript. HBB also analyzed the data and edited the manuscript. SPS supervised the work, helped in analyzing the data and edited the manuscript.
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Highlights
• Marine bacteria characterized on the basis of morphology, biochemical characters, antibiotic sensitivity and enzyme secretion
• Bacterial diversity assessed by phylogenetics and numerical taxonomy
• Stability of proteases at high temperatures, wide pH range and in Urea indicates their potential applications in detergent, leather, food, dairy, and cosmetics industries
• Salt dependent resistance against denaturants was revealed
• Study significantly adds to the diversity and enzymatic potential of the haloalkaliphilic bacteria of sea water
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Raval, V.H., Joshi, R.H., Bhatt, H.B. et al. Phenotypic characteristics, phylogenetic analysis and characterization of alkaline proteases of marine bacteria Geomicrobium halophilum, Oceanobacillus oncorhynchi, and Oceanobacillus khimchii. Biologia 77, 2405–2422 (2022). https://doi.org/10.1007/s11756-022-01095-7
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DOI: https://doi.org/10.1007/s11756-022-01095-7