Characterization of an extreme alkaline-stable keratinase from the draft genome of feather-degrading Bacillus sp.JM7 from deep-sea
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摘要
Bacillus sp. JM7, a strain isolated from the deep-sea of the South China Sea, was found to efficiently degrade 79.4% native chicken feather within 30 h. Scanning electron microscopy analysis showed that JM7 strain could gradually degrade feather by modifying the microstructure of feather keratin. A total of 25 protease genes were predicted from the draft genome of JM7 strain, among which a predicted subtilisin-like serine protease(designated as Ker02562) was further characterized for its keratinolytic activity. The recombinant Ker02562 functioned at a wide range of temperatures from 30℃ to 60℃, with an optimum at 40–50℃. Ker02562 was highly active at various pHs ranging from 5.0 to 13.0, with a maximum activity observed at pH 7.0–9.0. Remarkably, recombinant Ker02562 was stable in extreme alkaline environments(pH 10–13), which was much better than most other reported keratinases. Collectively, these favorable properties could make Bacillus sp. JM7 and Ker02562 attractive to be applied in the detergent formulation and feather bioconversion.
Bacillus sp. JM7, a strain isolated from the deep-sea of the South China Sea, was found to efficiently degrade 79.4% native chicken feather within 30 h. Scanning electron microscopy analysis showed that JM7 strain could gradually degrade feather by modifying the microstructure of feather keratin. A total of 25 protease genes were predicted from the draft genome of JM7 strain, among which a predicted subtilisin-like serine protease(designated as Ker02562) was further characterized for its keratinolytic activity. The recombinant Ker02562 functioned at a wide range of temperatures from 30℃ to 60℃, with an optimum at 40–50℃. Ker02562 was highly active at various pHs ranging from 5.0 to 13.0, with a maximum activity observed at pH 7.0–9.0. Remarkably, recombinant Ker02562 was stable in extreme alkaline environments(pH 10–13), which was much better than most other reported keratinases. Collectively, these favorable properties could make Bacillus sp. JM7 and Ker02562 attractive to be applied in the detergent formulation and feather bioconversion.
Bacillus sp. JM7, a strain isolated from the deep-sea of the South China Sea, was found to efficiently degrade 79.4% native chicken feather within 30 h. Scanning electron microscopy analysis showed that JM7 strain could gradually degrade feather by modifying the microstructure of feather keratin. A total of 25 protease genes were predicted from the draft genome of JM7 strain, among which a predicted subtilisin-like serine protease(designated as Ker02562) was further characterized for its keratinolytic activity. The recombinant Ker02562 functioned at a wide range of temperatures from 30℃ to 60℃, with an optimum at 40–50℃. Ker02562 was highly active at various pHs ranging from 5.0 to 13.0, with a maximum activity observed at pH 7.0–9.0. Remarkably, recombinant Ker02562 was stable in extreme alkaline environments(pH 10–13), which was much better than most other reported keratinases. Collectively, these favorable properties could make Bacillus sp. JM7 and Ker02562 attractive to be applied in the detergent formulation and feather bioconversion.
引文
Aehle W.2006.Enzymes in Industry:Products and Applications.2nd ed.Hoboken USA:John Wiley&Sons
Balaji S,Kumar M S,Karthikeyan R,et al.2008.Purification and characterization of an extracellular keratinase from a hornmeal-degrading Bacillus subtilis MTCC(9102).World Journal of Microbiology and Biotechnology,24:2741-2745,doi:10.1007/s11274-008-9782-7
Brandelli A.2008.Bacterial keratinases:useful enzymes for bioprocessing agroindustrial wastes and beyond.Food and Bioprocess Technology,1(2):105-116,doi:10.1007/s11947-007-0025-y
Brandelli A,Daroit D J,Riffel A.2010.Biochemical features of microbial keratinases and their production and applications.Applied Microbiology and Biotechnology,85(6):1735-1750,doi:10.1007/s00253-009-2398-5
Brandelli A,Riffel A.2005.Production of an extracellular keratinase from Chryseobacterium sp.growing on raw feathers.Electronic Journal of Biotechnology,8(1):35-42
Bressollier P,Letourneau F,Urdaci M,et al.1999.Purification and characterization of a keratinolytic serine proteinase from Streptomyces albidoflavus.Applied and Environmental Microbiology,65(6):2570-2576
Cao Zhangjun,Zhang Qi,Wei Dongkai,et al.2009.Characterization of a novel Stenotrophomonas isolate with high keratinase activity and purification of the enzyme.Journal of Industrial Microbiology and Biotechnology,36(2):181-188,doi:10.1007/s10295-008-0469-8
Corrêa A P F,Daroit D J,Brandelli A.2010.Characterization of a keratinase produced by Bacillus sp.P7 isolated from an Amazonian environment.International Biodeterioration and Biodegradation,64(1):1-6,doi:10.1016/j.ibiod.2009.06.015
da Gioppo N M R,Moreira-Gasparin F G,Costa A M,et al.2009.Influence of the carbon and nitrogen sources on keratinase production by Myrothecium verrucaria in submerged and solid state cultures.Journal of Industrial Microbiology and Biotechnology,36(5):705-711,doi:10.1007/s10295-009-0540-0
Delcher A L,Bratke K A,Powers E C,et al.2007.Identifying bacterial genes and endosymbiont DNA with Glimmer.Bioinformatics,23(6):673-679,doi:10.1093/bioinformatics/btm009
Farag A M,Hassan M A.2004.Purification,characterization and immobilization of a keratinase from Aspergillus oryzae.Enzyme and Microbial Technology,34(2):85-93,doi:10.1016/j.enzmictec.2003.09.002
Forgács G,Lundin M,Taherzadeh M J,et al.2013.Pretreatment of chicken feather waste for improved biogas production.Applied Biochemistry and Biotechnology,169(7):2016-2028,doi:10.1007/s12010-013-0116-3
Gao Chao,Jin Min,Yi Zhiwei,et al.2015.Characterization of a recombinant thermostable arylsulfatase from deep-sea bacterium Flammeovirga pacifica.Journal of Microbiology and Biotechnology,25:1894-1901,doi:10.4014/jmb.1504.04028
Goldstein J,Newbury D E,Echlin P,et al.1981.Scanning Electron Microscopy and X-ray Microanalysis:a Text for Biologists,Materials Scientists,and Geologists.US:Springer
Gong Jinsong,Wang Yue,Zhang Dandan,et al.2015.Biochemical characterization of an extreme alkaline and surfactant-stable keratinase derived from a newly isolated actinomycete Streptomyces aureofaciens K13.RSC Advances,5:24691-24699,doi:10.1039/C4RA16423G
Gradi?ar H,Kern S,Friedrich J.2000.Keratinase of Doratomyces microsporus.Applied Microbiology and Biotechnology,53(2):196-200,doi:10.1007/s002530050008
Gupta R,Sharma R,Beg Q K.2013.Revisiting microbial keratinases:next generation proteases for sustainable biotechnology.Critical Reviews in Biotechnology,33(2):216-228,doi:10.3109/07388551.2012.685051
Habbeche A,Saoudi B,Jaouadi B,et al.2014.Purification and biochemical characterization of a detergent-stable keratinase from a newly thermophilic actinomycete Actinomadura keratinilytica strain Cpt29 isolated from poultry compost.Journal of Bioscience and Bioengineering,117(4):413-421,doi:10.1016/j.jbiosc.2013.09.006
Jaouadi N Z,Rekik H,Badis A,et al.2013.Biochemical and molecular characterization of a serine keratinase from Brevibacillus brevis US575 with promising keratin-biodegradation and hidedehairing activities.PLoS One,8(10):e76722,doi:10.1371/journal.pone.0076722
Jeong J H,Jeon Y D,Lee O M,et al.2010.Characterization of a multifunctional feather-degrading Bacillus subtilis isolated from forest soil.Biodegradation,21(6):1029-1040,doi:10.1007/s10532-010-9363-y
Kublanov I V,Tsiroulnikov K B,Kaliberda E N,et al.2009.Keratinase of an anaerobic thermophilic bacterium Thermoanaerobacter sp.strain 1004-09 isolated from a hot spring in the Baikal rift zone.Microbiology,78(1):67-75,doi:10.1134/S0026261709010093
Lagesen K,Hallin P,R?dland E A,et al.2007.RNAmmer:consistent and rapid annotation of ribosomal RNA genes.Nucleic Acids Research,35(9):3100-3108,doi:10.1093/nar/gkm160
Li Ruiqiang,Li Yingrui,Kristiansen K,et al.2008.SOAP:short oligonucleotide alignment program.Bioinformatics,24(5):713-714,doi:10.1093/bioinformatics/btn025
Liu Qingyang,Zhang Tiehan,Song Nan,et al.2014.Purification and characterization of four key enzymes from a feather-degrading Bacillus subtilis from the gut of tarantula Chilobrachys guangxiensis.International Biodeterioration and Biodegradation,96:26-32,doi:10.1016/j.ibiod.2014.08.008
Nam G W,Lee D W,Lee H S,et al.2002.Native-feather degradation by Fervidobacterium islandicum AW-1,a newly isolated keratinase-producing thermophilic anaerobe.Archives of Microbiology,178(6):538-547,doi:10.1007/s00203-002-0489-0
Paul T,Das A,Mandal A,et al.2014.Production and purification of keratinase using chicken feather bioconversion by a newly isolated Aspergillus fumigatus TKF1:detection of valuable metabolites.Biomass Conversion and Biorefinery,4(2):137-148,doi:10.1007/s13399-013-0090-6
Schattner P,Brooks A N,Lowe T M.2005.The tRNAscan-SE,snoscan and snoGPS web servers for the detection of tRNAs and snoR-NAs.Nucleic Acids Research,33:686-689,doi:10.1093/nar/gki366
Suntornsuk W,Tongjun J,Onnim P,et al.2005.Purification and characterisation of keratinase from a thermotolerant feather-degrading bacterium.World Journal of Microbiology and Biotechnology,21:1111-1117,doi:10.1007/s11274-005-0078-x
Syed D G,Lee J C,Li Wenjun,et al.2009.Production,characterization and application of keratinase from Streptomyces gulbargensis.Bioresource Technology,100(5):1868-1871,doi:10.1016/j.biortech.2008.09.047
Thys R C S,Brandelli A.2006.Purification and properties of a keratinolytic metalloprotease from Microbacterium sp..Journal of Applied Microbiology,101(6):1259-1268,doi:10.1111/jam.2006.101.issue-6
Tiwary E,Gupta R.2010.Medium optimization for a novel 58kDa dimeric keratinase from Bacillus licheniformis ER-15:Biochemical characterization and application in feather degradation and dehairing of hides.Bioresource Technology,101(15):6103-6110,doi:10.1016/j.biortech.2010.02.090
Wang Ping,Wang Qiang,Cui Li,et al.2011.The combined use of cutinase,keratinase and protease treatments for wool bio-antifelting.Fibers and Polymers,12(6):760-764,doi:10.1007/s12221-011-0760-6
Zaghloul T I.1998.Cloned Bacillus subtilis alkaline protease(aprA)gene showing high level of keratinolytic activity.Applied Biochemistry and Biotechnology,70(1):199-205
Zhou Guilan,Jin Min,Cai Yaping,et al.2015.Characterization of a thermostable and alkali-stableα-amylase from deep-sea bacterium Flammeovirga pacifica.International Journal of Biological Macromolecules,80:676-682,doi:10.1016/j.ijbiomac.2015.07.042
Balaji S,Kumar M S,Karthikeyan R,et al.2008.Purification and characterization of an extracellular keratinase from a hornmeal-degrading Bacillus subtilis MTCC(9102).World Journal of Microbiology and Biotechnology,24:2741-2745,doi:10.1007/s11274-008-9782-7
Brandelli A.2008.Bacterial keratinases:useful enzymes for bioprocessing agroindustrial wastes and beyond.Food and Bioprocess Technology,1(2):105-116,doi:10.1007/s11947-007-0025-y
Brandelli A,Daroit D J,Riffel A.2010.Biochemical features of microbial keratinases and their production and applications.Applied Microbiology and Biotechnology,85(6):1735-1750,doi:10.1007/s00253-009-2398-5
Brandelli A,Riffel A.2005.Production of an extracellular keratinase from Chryseobacterium sp.growing on raw feathers.Electronic Journal of Biotechnology,8(1):35-42
Bressollier P,Letourneau F,Urdaci M,et al.1999.Purification and characterization of a keratinolytic serine proteinase from Streptomyces albidoflavus.Applied and Environmental Microbiology,65(6):2570-2576
Cao Zhangjun,Zhang Qi,Wei Dongkai,et al.2009.Characterization of a novel Stenotrophomonas isolate with high keratinase activity and purification of the enzyme.Journal of Industrial Microbiology and Biotechnology,36(2):181-188,doi:10.1007/s10295-008-0469-8
Corrêa A P F,Daroit D J,Brandelli A.2010.Characterization of a keratinase produced by Bacillus sp.P7 isolated from an Amazonian environment.International Biodeterioration and Biodegradation,64(1):1-6,doi:10.1016/j.ibiod.2009.06.015
da Gioppo N M R,Moreira-Gasparin F G,Costa A M,et al.2009.Influence of the carbon and nitrogen sources on keratinase production by Myrothecium verrucaria in submerged and solid state cultures.Journal of Industrial Microbiology and Biotechnology,36(5):705-711,doi:10.1007/s10295-009-0540-0
Delcher A L,Bratke K A,Powers E C,et al.2007.Identifying bacterial genes and endosymbiont DNA with Glimmer.Bioinformatics,23(6):673-679,doi:10.1093/bioinformatics/btm009
Farag A M,Hassan M A.2004.Purification,characterization and immobilization of a keratinase from Aspergillus oryzae.Enzyme and Microbial Technology,34(2):85-93,doi:10.1016/j.enzmictec.2003.09.002
Forgács G,Lundin M,Taherzadeh M J,et al.2013.Pretreatment of chicken feather waste for improved biogas production.Applied Biochemistry and Biotechnology,169(7):2016-2028,doi:10.1007/s12010-013-0116-3
Gao Chao,Jin Min,Yi Zhiwei,et al.2015.Characterization of a recombinant thermostable arylsulfatase from deep-sea bacterium Flammeovirga pacifica.Journal of Microbiology and Biotechnology,25:1894-1901,doi:10.4014/jmb.1504.04028
Goldstein J,Newbury D E,Echlin P,et al.1981.Scanning Electron Microscopy and X-ray Microanalysis:a Text for Biologists,Materials Scientists,and Geologists.US:Springer
Gong Jinsong,Wang Yue,Zhang Dandan,et al.2015.Biochemical characterization of an extreme alkaline and surfactant-stable keratinase derived from a newly isolated actinomycete Streptomyces aureofaciens K13.RSC Advances,5:24691-24699,doi:10.1039/C4RA16423G
Gradi?ar H,Kern S,Friedrich J.2000.Keratinase of Doratomyces microsporus.Applied Microbiology and Biotechnology,53(2):196-200,doi:10.1007/s002530050008
Gupta R,Sharma R,Beg Q K.2013.Revisiting microbial keratinases:next generation proteases for sustainable biotechnology.Critical Reviews in Biotechnology,33(2):216-228,doi:10.3109/07388551.2012.685051
Habbeche A,Saoudi B,Jaouadi B,et al.2014.Purification and biochemical characterization of a detergent-stable keratinase from a newly thermophilic actinomycete Actinomadura keratinilytica strain Cpt29 isolated from poultry compost.Journal of Bioscience and Bioengineering,117(4):413-421,doi:10.1016/j.jbiosc.2013.09.006
Jaouadi N Z,Rekik H,Badis A,et al.2013.Biochemical and molecular characterization of a serine keratinase from Brevibacillus brevis US575 with promising keratin-biodegradation and hidedehairing activities.PLoS One,8(10):e76722,doi:10.1371/journal.pone.0076722
Jeong J H,Jeon Y D,Lee O M,et al.2010.Characterization of a multifunctional feather-degrading Bacillus subtilis isolated from forest soil.Biodegradation,21(6):1029-1040,doi:10.1007/s10532-010-9363-y
Kublanov I V,Tsiroulnikov K B,Kaliberda E N,et al.2009.Keratinase of an anaerobic thermophilic bacterium Thermoanaerobacter sp.strain 1004-09 isolated from a hot spring in the Baikal rift zone.Microbiology,78(1):67-75,doi:10.1134/S0026261709010093
Lagesen K,Hallin P,R?dland E A,et al.2007.RNAmmer:consistent and rapid annotation of ribosomal RNA genes.Nucleic Acids Research,35(9):3100-3108,doi:10.1093/nar/gkm160
Li Ruiqiang,Li Yingrui,Kristiansen K,et al.2008.SOAP:short oligonucleotide alignment program.Bioinformatics,24(5):713-714,doi:10.1093/bioinformatics/btn025
Liu Qingyang,Zhang Tiehan,Song Nan,et al.2014.Purification and characterization of four key enzymes from a feather-degrading Bacillus subtilis from the gut of tarantula Chilobrachys guangxiensis.International Biodeterioration and Biodegradation,96:26-32,doi:10.1016/j.ibiod.2014.08.008
Nam G W,Lee D W,Lee H S,et al.2002.Native-feather degradation by Fervidobacterium islandicum AW-1,a newly isolated keratinase-producing thermophilic anaerobe.Archives of Microbiology,178(6):538-547,doi:10.1007/s00203-002-0489-0
Paul T,Das A,Mandal A,et al.2014.Production and purification of keratinase using chicken feather bioconversion by a newly isolated Aspergillus fumigatus TKF1:detection of valuable metabolites.Biomass Conversion and Biorefinery,4(2):137-148,doi:10.1007/s13399-013-0090-6
Schattner P,Brooks A N,Lowe T M.2005.The tRNAscan-SE,snoscan and snoGPS web servers for the detection of tRNAs and snoR-NAs.Nucleic Acids Research,33:686-689,doi:10.1093/nar/gki366
Suntornsuk W,Tongjun J,Onnim P,et al.2005.Purification and characterisation of keratinase from a thermotolerant feather-degrading bacterium.World Journal of Microbiology and Biotechnology,21:1111-1117,doi:10.1007/s11274-005-0078-x
Syed D G,Lee J C,Li Wenjun,et al.2009.Production,characterization and application of keratinase from Streptomyces gulbargensis.Bioresource Technology,100(5):1868-1871,doi:10.1016/j.biortech.2008.09.047
Thys R C S,Brandelli A.2006.Purification and properties of a keratinolytic metalloprotease from Microbacterium sp..Journal of Applied Microbiology,101(6):1259-1268,doi:10.1111/jam.2006.101.issue-6
Tiwary E,Gupta R.2010.Medium optimization for a novel 58kDa dimeric keratinase from Bacillus licheniformis ER-15:Biochemical characterization and application in feather degradation and dehairing of hides.Bioresource Technology,101(15):6103-6110,doi:10.1016/j.biortech.2010.02.090
Wang Ping,Wang Qiang,Cui Li,et al.2011.The combined use of cutinase,keratinase and protease treatments for wool bio-antifelting.Fibers and Polymers,12(6):760-764,doi:10.1007/s12221-011-0760-6
Zaghloul T I.1998.Cloned Bacillus subtilis alkaline protease(aprA)gene showing high level of keratinolytic activity.Applied Biochemistry and Biotechnology,70(1):199-205
Zhou Guilan,Jin Min,Cai Yaping,et al.2015.Characterization of a thermostable and alkali-stableα-amylase from deep-sea bacterium Flammeovirga pacifica.International Journal of Biological Macromolecules,80:676-682,doi:10.1016/j.ijbiomac.2015.07.042