Evaluation of the secondary structures of protein in the extracellular polymeric substances extracted from activated sludge by different methods
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摘要
The changes of protein secondary structures in the extracellular polymeric substances(EPS) extracted from activated sludge by four different methods were studied by analyzing the amide I region(1700–1600 cm-1) of the Fourier transform infrared spectra and model protein test. The results showed the molecular weight distribution of organic matter extracted by centrifugation, heating and cation exchange resin(CER) was similar, while the EPS extracted by centrifugation(Control) and CER had similar fluorescent organic matter. The protein secondary structures of extracted EPS by the four methods were different. The similarities of protein secondary structures between the EPS extracted by CER with the Control were the highest among the four extracted EPS. Although the EPS yield extracted by formaldehyde + NaOH method were the highest, its protein secondary structures had the lowest similarity with those extracted by the Control. Additionally, the effects of centrifugation and CER extraction on the secondary structures of bovine serum albumin were also lower than that of other extraction processes. CER enables the second maximum extraction of EPS and maximum retention of the original secondary structure of proteins.
The changes of protein secondary structures in the extracellular polymeric substances(EPS) extracted from activated sludge by four different methods were studied by analyzing the amide I region(1700–1600 cm-1) of the Fourier transform infrared spectra and model protein test. The results showed the molecular weight distribution of organic matter extracted by centrifugation, heating and cation exchange resin(CER) was similar, while the EPS extracted by centrifugation(Control) and CER had similar fluorescent organic matter. The protein secondary structures of extracted EPS by the four methods were different. The similarities of protein secondary structures between the EPS extracted by CER with the Control were the highest among the four extracted EPS. Although the EPS yield extracted by formaldehyde + NaOH method were the highest, its protein secondary structures had the lowest similarity with those extracted by the Control. Additionally, the effects of centrifugation and CER extraction on the secondary structures of bovine serum albumin were also lower than that of other extraction processes. CER enables the second maximum extraction of EPS and maximum retention of the original secondary structure of proteins.
The changes of protein secondary structures in the extracellular polymeric substances(EPS) extracted from activated sludge by four different methods were studied by analyzing the amide I region(1700–1600 cm-1) of the Fourier transform infrared spectra and model protein test. The results showed the molecular weight distribution of organic matter extracted by centrifugation, heating and cation exchange resin(CER) was similar, while the EPS extracted by centrifugation(Control) and CER had similar fluorescent organic matter. The protein secondary structures of extracted EPS by the four methods were different. The similarities of protein secondary structures between the EPS extracted by CER with the Control were the highest among the four extracted EPS. Although the EPS yield extracted by formaldehyde + NaOH method were the highest, its protein secondary structures had the lowest similarity with those extracted by the Control. Additionally, the effects of centrifugation and CER extraction on the secondary structures of bovine serum albumin were also lower than that of other extraction processes. CER enables the second maximum extraction of EPS and maximum retention of the original secondary structure of proteins.
引文
APHA(Ed.),1998.Standard Methods for the Examination of Water and Wastewater,20th ed APHA,Washington,DC,USA.
Avitabile,C.,D'Andrea,L.D.,Romanelli,A.,2014.Circular dichroism studies on the interactions of antimicrobial peptides with bacterial cells.Sci.Rep.4,4293.
Bourven,I.,Simon,S.,Guibaud,G.,2013.Influence of extraction method on size exclusion chromatography fingerprints of EPS from wastewater sludges.Environ.Technol.34(1-4),321-332.
Chabaliná,L.D.,Pastor,M.R.,Rico,D.P.,2013.Characterization of soluble and bound EPS obtained from 2 submerged membrane bioreactors by 3D-EEM and HPSEC.Talanta 115,706-712.
Chen,W.,Westerhoff,P.,Leenheer,J.A.,Booksh,K.,2003.Fluorescence excitation-emission matrix regional integration to quantify spectra for dissolved organic matter.Environ.Sci.Technol.37,5701-5710.
Chodankar,S.,Aswal,V.K.,Kohlbrecher,J.,Vavrin,R.,Wagh,A.G.,2008.Structural evolution during protein denaturation as induced by different methods.Phys.Rev.E 77,031901.
Drozdetskiy,A.,Cole,C.,Procter,J.,Barton,G.J.,2015.JPred4:a protein secondary structure prediction server.Nucleic Acids Res.43(W1),W389-W394.
Dubois,M.,Gilles,K.A.,Hamilton,J.K.,Rebers,P.A.,Smith,F.,1956.Calorimetric method for determination of sugars and related substances.Anal.Chem.28,350-356.
Feng,L.J.,Wang,J.J.,Liu,S.C.,Sun,X.D.,Yuan,X.Z.,Wang,S.G.,2018.Role of extracellular polymeric substances in the acute inhibition of activated sludge by polystyrene nanoparticles.Environ.Pollut.238,859-865.
Güler,G.,Vorob'ev,M.M.,Vogel,V.,M?ntele,W.,2016.Proteolytically-induced changes of secondary structural protein conformation of bovine serum albumin monitored by Fourier transform infrared(FT-IR)and UV-circular dichroism spectroscopy.Spectrochim.Acta A 161,8-18.
Guo,X.,Liu,J.,Xiao,B.,2014.Evaluation of the damage of cell wall and cell membrane for various extracellular polymeric substance extractions of activated sludge.J.Biotech.188,130-135.
Hong,P.N.,Honda,R.,Noguchi,M.,Ito,T.,2017.Optimum selection of extraction methods of extracellular polymeric substances in activated sludge for effective extraction of the target components.Biochem.Eng.J.127,136-146.
Kavitha,S.,Kumar,S.A.,Yogalakshmi,K.N.,Kaliappan,S.,Banu,J.R.,2013.Effect of enzyme secreting bacterial pretreatment on Enhancement of aerobic digestion potential of WAS interceded through EDTA.Bioresour.Technol.150,210-219.
Kavitha,S.,Kumar,S.A.,Kaliappan,S.,Yeom,I.T.,Banu,J.R.,2014a.Improving the amenability of municipal waste activated sludge for biological pretreatment by phase-separated sludge disintegration method.Bioresour.Technol.164,700-706.
Kavitha,S.,Jayashree,C.,Kumar,S.A.,Yeom,I.T.,Banu,J.R.,2014b.The enhancement of anaerobic biodegradability of waste activated sludge by surfactant mediated biological pretreatment.Bioresour.Technol.168,159-166.
Kavitha,S.,Kaliappan,S.,Kumar,S.A.,Yeom,I.T.,Banu,J.R.,2015a.Effect of NaCl induced floc disruption on biological disintegration of sludge for enhanced biogas production.Bioresour.Technol.192,807-811.
Kavitha,S.,Kumar,S.A.,Kaliappan,S.,Yeom,I.T.,Banu,J.R.,2015b.Achieving profitable biological sludge disintegration through phase separation and predicting its anaerobic biodegradability by non linear regression modelling.Chem.Eng.J.279,478-487.
Kwaambwa,H.M.,Maikokera,R.,2008.Infrared and circular dichroism spectroscopic characterization of secondary structure components of a water treatment coagulant protein extracted from Moringa oleifera seeds.Colloid Surf.64,118-125.
Li,R.,Wu,Z.,Wang,Y.,Ding,L.,Wang,Y.,2016.Role of pH-induced structural change in protein aggregation in foam fractionation of bovine serum albumin.Biotechnol.Rep.9,46-52.
Liu,H.,Fang,H.H.P.,2002.Extraction of extracellular polymeric substances(EPS)of sludges.J.Biotechnol.95,249-256.
Lowry,N.J.,Rosebrough,A.L.,Farr Randall,R.J.,1951.Protein measurement with the Folin phenol reagent.J.Biol.Chem.1993,265-275.
Lu,Q.,Chang,M.,Yu,Z.,Zhou,S.,2016.The effects of three commonly used extraction methods on the redox properties of extracellular polymeric substances from activated sludge.Environ.Technol.36,2884-2891.
Luo,K.,Yang,Q.,Li,Q.M.,Chen,H.B.,Zeng,G.M.,2013.Novel insights into enzymatic-enhanced anaerobic digestion of waste activated sludge by three-dimensional excitation and emission matrix fluorescence spectroscopy.Chemosphere 91,579-585.
Matsuo,K.,Sakurada,Y.,Yonehara,R.,Kataoka,M.,Gekko,K.,2007.Secondary-structure analysis of denatured proteins by vacuum-ultraviolet circular dichroism spectroscopy.Biophys.J.92,4088-4096.
Nouha,K.,Kumar,R.S.,Balasubramanian,S.,Tyaji,R.D.,2018.Critical review of EPS production,synthesis and composition for sludge flocculation.J.Environ.Sci.66,225-245.
Perczel,A.,Park,K.,Fasman,G.,1992.Analysis of the circular dichroism spectrum of proteins using the convex constraint algorithm:a practical guide.Anal.Biochem.203(1),83-93.
R Development Core Team,2015.R:A Language and Environment for Statistical Computing Vienna,Austria.
Sheng,G.P.,Yu,H.Q.,2006.Characterization of extracellular polymeric substances of aerobic and anaerobic sludge using three-dimensional excitation and emission matrix fluorescence spectroscopy.Water Res.40,1233-1239.
Sheng,G.P.,Yu,H.Q.,Li,X.Y.,2010.Extracellular polymeric substances(EPS)of microbial aggregates in biological wastewater treatment systems:a review.Biotechnol.Adv.28,882-894.
Simon,S.,Pa?ro,B.,Villain,M.,D'Abzac,P.,Van Hullebusch,E.,Lens,P.,2009.Evaluation of size exclusion chromatography(SEC)for the characterization of extracellular polymeric substances(EPS)in anaerobic granular sludges.Bioresour.Technol.100,6258-6268.
Stuart,B.,2004.Infrared Spectroscopy:Fundamentals and Applications.John Wiley&Sons.
Sun,Y.,Clinkenbeard,K.D.,Clarke,C.,Cudd,L.,Highlander,S.K.,Dabo,S.M.,1999.Pasteurella haemolytica leukotoxin induced apoptosis of bovine lymphocytes involves DNA fragmentation.Vet.Microbiol.65,153-166.
Wang,Z.W.,Wu,Z.C.,Tang,S.J.,2009.Characterization of dissolved organic matter in a submerged membrane bioreactor by using three-dimensional excitation and emission matrix fluorescence spectroscopy.Water Res.43,1533-1540.
Wang,S.,Peng,J.,Ma,J.,Xu,J.,2016a.Protein secondary structure prediction using deep convolutional neural fields.Sci.Reports.6,18962.
Wang,X.,Li,Y.,Liu,J.,Ren,N.Q.,Qu,J.,2016b.Augmentation of protein-derived acetic acid production by heat-alkalineinduced changes in protein structure and conformation.Water Res.88,595-603.
Wang,B.B.,Liu,X.T.,Chen,J.M.,Peng,D.C.,He,F.,2018.Composition and functional group characterization of extracellular polymeric substances(EPS)in activated sludge:the impacts of polymerization degree of proteinaceous substrates.Water Res.129,133-142.
Wu,B.,Ni,B.J.,Horvat,K.,Song,L.,Chai,X.,Dai,X.,et al.,2017.Occurrence state and molecular structure analysis of extracellular proteins with implications on the dewaterability of waste-activated sludge.Environ.Sci.Technol.51,9235-9243.
Xiao,N.,Chen,Y.,Chen,A.,Feng,L.,2014.Enhanced bio-hydrogen production from protein wastewater by altering protein structure and amino acids acidification type.Sci.Rep.4,3992.
Xiao,B.,Liu,C.,Jun,X.,Guo,X.,2015.Evaluation of the microbial cell structure damages in alkaline pretreatment of waste activated sludge.Bioresour.Technol.196,109-115.
Xie,M.,Wang,C.,Liu,X.,Xiong,R.,Xu,Y.,2017.Characteristics of biochemical and fractal structure of activated sludge with thermochemical lysis.Water Air Soil Poll.228,187.
Yin,C.,Meng,F.,Chen,G.-H.,2015.Spectroscopic characterization of extracellular polymeric substances from a mixed culture dominated by ammonia-oxidizing bacteria.Water Res.68,740-749.
You,G.,Wang,P.,Hou,J.,Wang,C.,Xu,Y.,Miao,L.,et al.,2017.Insights into the short-term effects of Ce O2nanoparticles on sludge dewatering and related mechanism.Water Res.118,93-103.
Zhang,L.,Feng,X.,Zhu,N.,Chen,J.,2007.Role of extracellular protein in the formation and stability of aerobic granules.Enzym.Microb.Technol.41,551-557.
Zhong,K.,Wu,J.,Wang,Z.,Chen,F.,Liao,X.,Hu,X.,et al.,2007.Inactivation kinetics and secondary structural change of PEF-treated POD and PPO.Food Chem.100(1),115-123.
Zhou,J.,Wang,J.J.,Baudon,A.,Chow,A.T.,2013.Improved fluorescence excitation-emission matrix regional integration to quantify spectra for fluorescent dissolved organic matter.J.Environ.Qual.42,925-930.
Zhu,L.,Yu,H.,Liu,Y.,Qi,H.,Xu,X.,2015.Optimization for extracellular polymeric substances extraction of microbial aggregates.Water Sci.Technol.71,1106-1112.
Avitabile,C.,D'Andrea,L.D.,Romanelli,A.,2014.Circular dichroism studies on the interactions of antimicrobial peptides with bacterial cells.Sci.Rep.4,4293.
Bourven,I.,Simon,S.,Guibaud,G.,2013.Influence of extraction method on size exclusion chromatography fingerprints of EPS from wastewater sludges.Environ.Technol.34(1-4),321-332.
Chabaliná,L.D.,Pastor,M.R.,Rico,D.P.,2013.Characterization of soluble and bound EPS obtained from 2 submerged membrane bioreactors by 3D-EEM and HPSEC.Talanta 115,706-712.
Chen,W.,Westerhoff,P.,Leenheer,J.A.,Booksh,K.,2003.Fluorescence excitation-emission matrix regional integration to quantify spectra for dissolved organic matter.Environ.Sci.Technol.37,5701-5710.
Chodankar,S.,Aswal,V.K.,Kohlbrecher,J.,Vavrin,R.,Wagh,A.G.,2008.Structural evolution during protein denaturation as induced by different methods.Phys.Rev.E 77,031901.
Drozdetskiy,A.,Cole,C.,Procter,J.,Barton,G.J.,2015.JPred4:a protein secondary structure prediction server.Nucleic Acids Res.43(W1),W389-W394.
Dubois,M.,Gilles,K.A.,Hamilton,J.K.,Rebers,P.A.,Smith,F.,1956.Calorimetric method for determination of sugars and related substances.Anal.Chem.28,350-356.
Feng,L.J.,Wang,J.J.,Liu,S.C.,Sun,X.D.,Yuan,X.Z.,Wang,S.G.,2018.Role of extracellular polymeric substances in the acute inhibition of activated sludge by polystyrene nanoparticles.Environ.Pollut.238,859-865.
Güler,G.,Vorob'ev,M.M.,Vogel,V.,M?ntele,W.,2016.Proteolytically-induced changes of secondary structural protein conformation of bovine serum albumin monitored by Fourier transform infrared(FT-IR)and UV-circular dichroism spectroscopy.Spectrochim.Acta A 161,8-18.
Guo,X.,Liu,J.,Xiao,B.,2014.Evaluation of the damage of cell wall and cell membrane for various extracellular polymeric substance extractions of activated sludge.J.Biotech.188,130-135.
Hong,P.N.,Honda,R.,Noguchi,M.,Ito,T.,2017.Optimum selection of extraction methods of extracellular polymeric substances in activated sludge for effective extraction of the target components.Biochem.Eng.J.127,136-146.
Kavitha,S.,Kumar,S.A.,Yogalakshmi,K.N.,Kaliappan,S.,Banu,J.R.,2013.Effect of enzyme secreting bacterial pretreatment on Enhancement of aerobic digestion potential of WAS interceded through EDTA.Bioresour.Technol.150,210-219.
Kavitha,S.,Kumar,S.A.,Kaliappan,S.,Yeom,I.T.,Banu,J.R.,2014a.Improving the amenability of municipal waste activated sludge for biological pretreatment by phase-separated sludge disintegration method.Bioresour.Technol.164,700-706.
Kavitha,S.,Jayashree,C.,Kumar,S.A.,Yeom,I.T.,Banu,J.R.,2014b.The enhancement of anaerobic biodegradability of waste activated sludge by surfactant mediated biological pretreatment.Bioresour.Technol.168,159-166.
Kavitha,S.,Kaliappan,S.,Kumar,S.A.,Yeom,I.T.,Banu,J.R.,2015a.Effect of NaCl induced floc disruption on biological disintegration of sludge for enhanced biogas production.Bioresour.Technol.192,807-811.
Kavitha,S.,Kumar,S.A.,Kaliappan,S.,Yeom,I.T.,Banu,J.R.,2015b.Achieving profitable biological sludge disintegration through phase separation and predicting its anaerobic biodegradability by non linear regression modelling.Chem.Eng.J.279,478-487.
Kwaambwa,H.M.,Maikokera,R.,2008.Infrared and circular dichroism spectroscopic characterization of secondary structure components of a water treatment coagulant protein extracted from Moringa oleifera seeds.Colloid Surf.64,118-125.
Li,R.,Wu,Z.,Wang,Y.,Ding,L.,Wang,Y.,2016.Role of pH-induced structural change in protein aggregation in foam fractionation of bovine serum albumin.Biotechnol.Rep.9,46-52.
Liu,H.,Fang,H.H.P.,2002.Extraction of extracellular polymeric substances(EPS)of sludges.J.Biotechnol.95,249-256.
Lowry,N.J.,Rosebrough,A.L.,Farr Randall,R.J.,1951.Protein measurement with the Folin phenol reagent.J.Biol.Chem.1993,265-275.
Lu,Q.,Chang,M.,Yu,Z.,Zhou,S.,2016.The effects of three commonly used extraction methods on the redox properties of extracellular polymeric substances from activated sludge.Environ.Technol.36,2884-2891.
Luo,K.,Yang,Q.,Li,Q.M.,Chen,H.B.,Zeng,G.M.,2013.Novel insights into enzymatic-enhanced anaerobic digestion of waste activated sludge by three-dimensional excitation and emission matrix fluorescence spectroscopy.Chemosphere 91,579-585.
Matsuo,K.,Sakurada,Y.,Yonehara,R.,Kataoka,M.,Gekko,K.,2007.Secondary-structure analysis of denatured proteins by vacuum-ultraviolet circular dichroism spectroscopy.Biophys.J.92,4088-4096.
Nouha,K.,Kumar,R.S.,Balasubramanian,S.,Tyaji,R.D.,2018.Critical review of EPS production,synthesis and composition for sludge flocculation.J.Environ.Sci.66,225-245.
Perczel,A.,Park,K.,Fasman,G.,1992.Analysis of the circular dichroism spectrum of proteins using the convex constraint algorithm:a practical guide.Anal.Biochem.203(1),83-93.
R Development Core Team,2015.R:A Language and Environment for Statistical Computing Vienna,Austria.
Sheng,G.P.,Yu,H.Q.,2006.Characterization of extracellular polymeric substances of aerobic and anaerobic sludge using three-dimensional excitation and emission matrix fluorescence spectroscopy.Water Res.40,1233-1239.
Sheng,G.P.,Yu,H.Q.,Li,X.Y.,2010.Extracellular polymeric substances(EPS)of microbial aggregates in biological wastewater treatment systems:a review.Biotechnol.Adv.28,882-894.
Simon,S.,Pa?ro,B.,Villain,M.,D'Abzac,P.,Van Hullebusch,E.,Lens,P.,2009.Evaluation of size exclusion chromatography(SEC)for the characterization of extracellular polymeric substances(EPS)in anaerobic granular sludges.Bioresour.Technol.100,6258-6268.
Stuart,B.,2004.Infrared Spectroscopy:Fundamentals and Applications.John Wiley&Sons.
Sun,Y.,Clinkenbeard,K.D.,Clarke,C.,Cudd,L.,Highlander,S.K.,Dabo,S.M.,1999.Pasteurella haemolytica leukotoxin induced apoptosis of bovine lymphocytes involves DNA fragmentation.Vet.Microbiol.65,153-166.
Wang,Z.W.,Wu,Z.C.,Tang,S.J.,2009.Characterization of dissolved organic matter in a submerged membrane bioreactor by using three-dimensional excitation and emission matrix fluorescence spectroscopy.Water Res.43,1533-1540.
Wang,S.,Peng,J.,Ma,J.,Xu,J.,2016a.Protein secondary structure prediction using deep convolutional neural fields.Sci.Reports.6,18962.
Wang,X.,Li,Y.,Liu,J.,Ren,N.Q.,Qu,J.,2016b.Augmentation of protein-derived acetic acid production by heat-alkalineinduced changes in protein structure and conformation.Water Res.88,595-603.
Wang,B.B.,Liu,X.T.,Chen,J.M.,Peng,D.C.,He,F.,2018.Composition and functional group characterization of extracellular polymeric substances(EPS)in activated sludge:the impacts of polymerization degree of proteinaceous substrates.Water Res.129,133-142.
Wu,B.,Ni,B.J.,Horvat,K.,Song,L.,Chai,X.,Dai,X.,et al.,2017.Occurrence state and molecular structure analysis of extracellular proteins with implications on the dewaterability of waste-activated sludge.Environ.Sci.Technol.51,9235-9243.
Xiao,N.,Chen,Y.,Chen,A.,Feng,L.,2014.Enhanced bio-hydrogen production from protein wastewater by altering protein structure and amino acids acidification type.Sci.Rep.4,3992.
Xiao,B.,Liu,C.,Jun,X.,Guo,X.,2015.Evaluation of the microbial cell structure damages in alkaline pretreatment of waste activated sludge.Bioresour.Technol.196,109-115.
Xie,M.,Wang,C.,Liu,X.,Xiong,R.,Xu,Y.,2017.Characteristics of biochemical and fractal structure of activated sludge with thermochemical lysis.Water Air Soil Poll.228,187.
Yin,C.,Meng,F.,Chen,G.-H.,2015.Spectroscopic characterization of extracellular polymeric substances from a mixed culture dominated by ammonia-oxidizing bacteria.Water Res.68,740-749.
You,G.,Wang,P.,Hou,J.,Wang,C.,Xu,Y.,Miao,L.,et al.,2017.Insights into the short-term effects of Ce O2nanoparticles on sludge dewatering and related mechanism.Water Res.118,93-103.
Zhang,L.,Feng,X.,Zhu,N.,Chen,J.,2007.Role of extracellular protein in the formation and stability of aerobic granules.Enzym.Microb.Technol.41,551-557.
Zhong,K.,Wu,J.,Wang,Z.,Chen,F.,Liao,X.,Hu,X.,et al.,2007.Inactivation kinetics and secondary structural change of PEF-treated POD and PPO.Food Chem.100(1),115-123.
Zhou,J.,Wang,J.J.,Baudon,A.,Chow,A.T.,2013.Improved fluorescence excitation-emission matrix regional integration to quantify spectra for fluorescent dissolved organic matter.J.Environ.Qual.42,925-930.
Zhu,L.,Yu,H.,Liu,Y.,Qi,H.,Xu,X.,2015.Optimization for extracellular polymeric substances extraction of microbial aggregates.Water Sci.Technol.71,1106-1112.