Volume 11 Issue 1
Jan.  2021
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Article Navigation >  Journal of Environmental Engineering Technology  > 2021  >  11(1): 144-150
YANG Kai, ZHANG Shuo, CUI Kangping, HEI Shengqiang, LÜ Xuemin, SONG Guangqing, ZHANG Xian, HUANG Xia. Evaluation of protein quantification methods for extracellular polymers in activated sludge[J]. Journal of Environmental Engineering Technology, 2021, 11(1): 144-150. doi: 10.12153/j.issn.1674-991X.20200046
Citation: YANG Kai, ZHANG Shuo, CUI Kangping, HEI Shengqiang, LÜ Xuemin, SONG Guangqing, ZHANG Xian, HUANG Xia. Evaluation of protein quantification methods for extracellular polymers in activated sludge[J]. Journal of Environmental Engineering Technology, 2021, 11(1): 144-150. doi: 10.12153/j.issn.1674-991X.20200046
shu

Evaluation of protein quantification methods for extracellular polymers in activated sludge

doi: 10.12153/j.issn.1674-991X.20200046
  • 1.

    School of Resources and Environmental Engineering,Hefei University of Technology

  • 2.

    State Key Laboratory of Environmental Simulation and Pollution Control, School of Environment, Tsinghua University

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  • Corresponding author: HUANG Xia E-mail: xhuang@tsinghua.edu.cn
  • Received Date: 2020-03-09
  • Publish Date: 2021-01-20
    Abstract
  • The accurate quantification of proteins in extracellular polymeric substances (EPS) is of great significance for studying the properties of activated sludge. To provide a basis for the selection of protein quantification methods in EPS, the test performance of Modified Lowry method, BCA method, Bradford method was systematically evaluated, including the test range, accuracy, precision and the anti-interference of some components (Ca2+, Mg2+,reducing sugar and humic acid) in EPS. The results indicated that the Modified Lowry method showed a good linear correlation between protein concentration and absorbance in the range of protein concentration of 1-10 and 10-100 mg/L (R2>0.99); and the test range of BCA method and Bradford method was wider. Modified Lowry method had the highest accuracy and precision, BCA method ranked the second, and Bradford method was the worst. Besides, Modified Lowry method was affected by Ca2+, Mg2+and reducing sugar; BCA method was affected by reducing sugar and humic acid; and Bradford method was affected by humic acid. Therefore, the choice of protein quantification method in EPS should be comprehensively considered from the two aspects of sample properties (protein content range, interference species and concentration) and test method performance (test range, accuracy, precision and anti-interference).

     

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    • References(29)
  • [1]
    KAVITHA S, JAYASHREE C, ADISH K S, et al. The enhancement of anaerobic biodegradability of waste activated sludge by surfactant mediated biological pretreatment[J]. Bioresource Technology, 2014,168(3):159-166.
    [2]
    WANG B B, LIU X T, CHEN J M, et al. Composition and functional group characterization of extracellular polymeric substances (EPS) in activated sludge:the impacts of polymerization degree of proteinaceous substrates[J]. Water Research, 2018,129:133-142.
    pmid: 29145083
    [3]
    CHEN L, TIAN Y, CAO C Q, et al. Interaction energy evaluation of soluble microbial products (SMP) on different membrane surfaces:role of the reconstructed membrane topology[J]. Water Research, 2012,46(8):2693-2704.
    pmid: 22406287
    [4]
    NI B J, RITTMANN B E, YU H Q. Soluble microbial products and their implications in mixed culture biotechnology[J]. Trends in Biotechnology, 2011,29(9):454-463.
    pmid: 21632131
    [5]
    KUNACHEVA C, STUCKEY D C. Analytical methods for soluble microbial products (SMP) and extracellular polymers (ECP) in wastewater treatment systems:a review[J]. Water Research, 2014,61(Suppl):1-18.
    [6]
    ZHANG P, SHEN Y, GUO J S, et al. Extracellular protein analysis of activated sludge and their functions in wastewater treatment plant by shotgun proteomics[J]. Scientific Reports, 2015,5:12041.
    doi: 10.1038/srep12041 pmid: 26160685
    [7]
    HOU X, LIU S, ZHANG Z. Role of extracellular polymeric substance in determining thehigh aggregation ability of anammox sludge[J]. Water Research, 2015,75:51-62.
    doi: 10.1016/j.watres.2015.02.031 pmid: 25746962
    [8]
    YIN C, MENG F, CHEN G H. Spectroscopic characterization of extracellular polymeric substances from a mixed culture dominated by ammonia-oxidizing bacteria[J]. Water Research, 2015,68:740-749.
    doi: 10.1016/j.watres.2014.10.046 pmid: 25462778
    [9]
    WU B, NI B J, HORVAT K, et al. Occurrence state and molecular structure analysis of extracellular proteins with implications to the dewaterability of waste activated sludge[J]. Environmental Science & Technology, 2017,51(16):9235-9243.
    doi: 10.1021/acs.est.7b02861 pmid: 28741346
    [10]
    FELIPE M O, LEMA J, RAMON M, et al. Role of exopolymeric protein on the settleability of nitrifying sludges[J]. Bioresource Technology, 2004,94(1):43-48.
    doi: 10.1016/j.biortech.2003.11.010 pmid: 15081485
    [11]
    XIAO B Y, LIU Y, LUO M, et al. Evaluation of the secondary structures of protein in the extracellular polymeric substances extracted from activated sludge by different methods[J]. Journal of Environmental Sciences, 2019(6):128-136.
    [12]
    BRADSTREET R B. Kjeldahl method for organic nitrogen[J]. Analytical Chemistry, 1954,26(1):185-187.
    [13]
    HIRT R C. Ultraviolet spectrophotometry[J]. Analytical Chemistry, 1956,28(4):579-583.
    [14]
    GORNALL A G, BARDWILL C J, DAVID M N. Determination of serum protein by means of the Biuret reaction[J]. Journal of Biological Chemistry, 1949,177:751-766.
    [15]
    FRØLUND B, GRIEBE T, NIELSEN P H. Enzymatic-activity in the activated-sludge floc matrix[J]. Applied Microbiology and Biotechnology, 1995,43(4):755-761.
    doi: 10.1007/BF00164784 pmid: 7546613
    [16]
    LOWRY O H, ROSEBROUGH N J, FARR A L, et al. Protein measurement with the Folin phenol reagent[J]. Journal of Biological Chemistry, 1951,193(1):265-275.
    [17]
    SMITH P K, KROHN R I, HERMANSON G T. Measurement of protein using bicinchoninic acid[J]. Analytical Biochemistry, 1985,163(1):76-85.
    [18]
    BRANFORD M M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding[J]. Analytical Biochemistry, 1976,72(1/2):248-254.
    [19]
    YUAN H X, QIN F, GUO W Q, et al. Oxidative stress and spermatogenesis suppression in the testis of cadmium-treated Bombyx mori larvae[J]. Environmental Science and Pollution Research, 2015,23(6):5763-5770.
    doi: 10.1007/s11356-015-5818-3 pmid: 26585454
    [20]
    DONG Q, PARKER W, DAGNEW M. Impact of FeCl3,dosing on AnMBR treatment of municipal wastewater[J]. Water Research, 2015,80:281-293.
    doi: 10.1016/j.watres.2015.04.025 pmid: 26005788
    [21]
    KASZYCKI P, DUBICKA-LISOWSKA A, AUGUSTYNOWICZ J, et al. Callitriche cophocarpa (water starwort) proteome under chromate stress:evidence for induction of a quinone reductase[J]. Environmental Science and Pollution Research, 2018,25(10):1-15.
    [22]
    SHEN Y X, XIAO K, LIANG P, et al. Improvement on the modified Lowry method against interference of divalent cations in soluble protein measurement[J]. Applied Microbiology and Biotechnology, 2013,97(9):4167-4178.
    doi: 10.1007/s00253-013-4783-3 pmid: 23474613
    [23]
    FELZ S, VERMEULEN P, van LOOSDRECHT M C M, et al. Chemical characterization methods for the analysis of structural extracellular polymeric substances (EPS)[J]. Water Research, 2019,157(15):201-208.
    [24]
    LE C C, KUNACHEVA C, STUCKEY D C. “Protein” measurement in biological wastewater treatment systems:a critical evaluation[J]. Environmental Science & Technology, 2016,50(6):3074-3081.
    doi: 10.1021/acs.est.5b05261 pmid: 26893149
    [25]
    MORENO M R D, SMITH J F, SMITH R V. Mechanism studies of coomassie blue and silver staining of proteins[J]. Journal of Pharmaceutical Sciences, 2010,75(9):907-911.
    doi: 10.1002/jps.2600750919 pmid: 2431134
    [26]
    HENRIST C, MATHIEU J P, VOGELS C, et al. Morphological study of magnesium hydroxide nanoparticles precipitated in dilute aqueous solution[J]. Journal of Crystal Growth, 2003,249(1/2):321-330.
    [27]
    LÜ J, QIU L, QU B. Controlled growth of three morphological structures of magnesium hydroxide nanoparticles by wet precipitation method[J]. Journal of Crystal Growth, 2004,267(3/4):676-684.
    [28]
    WAFFENSCHMIDT S, JAENICKE L. Assay of reducing sugars in the nanomole range with 2,2’-bicinchoninate[J]. Analytical Biochemistry, 1987,165(2):337-340.
    doi: 10.1016/0003-2697(87)90278-8 pmid: 3425902
    [29]
    刘新超, 李俊, 谢丽, 等. 腐殖酸表征方法研究进展[J]. 净水技术, 2009,28(3):6-9.
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