Recovery of cyanophycin granule polypeptide from activated sludge: carbon source dependence and aggregation-induced luminescence characteristics

doi:10.1007/s11783-024-1776-7

Front. Environ. Sci. Eng.

2024, Vol. 18

Issue (2) : 16 https://doi.org/10.1007/s11783-024-1776-7

RESEARCH ARTICLE

Recovery of cyanophycin granule polypeptide from activated sludge: carbon source dependence and aggregation-induced luminescence characteristics

Kui Zou, Hongyuan Liu, Bo Feng, Taiping Qing, Peng Zhang(

)

College of Environment and Resources, Xiangtan University, Xiangtan 411105, China

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Abstract

● Sodium acetate significantly enriched the CGP synthetase-encoding gene.

● The highest CGP yield was obtained from activated sludge fed with sodium acetate.

● Biofilm is more conducive to CGP accumulation compared with floc sludge.

● Aggregation-induced luminescence of CGP was first reported.

In the sewage treatment process, facilitating the conversion of pollutants into value-added resources holds great potential for reducing the amount of greenhouse gas emissions and promoting economic circulation. Cyanophycin granule polypeptide (CGP), a recently discovered high value-added biopolymer present in activated sludge, has provided new avenues for the recovery of resources. However, the mechanisms that regulate CGP synthesis and the characteristics of this biopolymer in activated sludge remain unclear thus far. This study investigated the synthesis of CGP, polyhydroxyalkanoates (PHA), and alginate-like exopolysaccharides (ALE) in various microbial aggregates under different carbon sources feeding conditions. Our results showed that the CGP yields was superior that of PHA and ALE when subjected to identical carbon source feeding conditions. Furthermore, biofilm was more conducive to CGP accumulation than floc sludge. Compared with glucose and methanol, sodium acetate significantly enriched the CGP synthetase-encoding gene (cphA_abundance = ~17419), resulting in the highest CGP yield (average 107.1 mg/g MLSS) in both biofilm and floc sludge. This study is the first to reported the characteristic fluorescence of CGP (Ex/Em = ~360/450 nm) caused by the aggregated luminescence of arginine on the side chains. Overall, this study highlights the potential application of CGP as a fluorescent material and offers insights into CGP recovery from activated sludge in wastewater treatment plants.

Keywords Cyanophycin Polyhydroxyalkanoate Alginate-like exopolysaccharides Bioresource recovery Aggregation-induced luminescence

Corresponding Author(s): Peng Zhang

Issue Date: 18 September 2023

Cite this article:

Kui Zou,Hongyuan Liu,Bo Feng, et al. Recovery of cyanophycin granule polypeptide from activated sludge: carbon source dependence and aggregation-induced luminescence characteristics[J]. Front. Environ. Sci. Eng., 2024, 18(2): 16.

URL:

https://academic.hep.com.cn/fese/EN/10.1007/s11783-024-1776-7
https://academic.hep.com.cn/fese/EN/Y2024/V18/I2/16

Fig.1 Water quality and biopolymer yields in biofilm and floc sludge reactors fed with different carbon sources. (a) Effluent quality during reactor operation, (b) difference analysis of PHA, CGP, and ALE yields, (c) total biopolymer yields in floc sludge and biofilm.

Fig.2 FTIR spectra of three biopolymers (a) CGP, (b) PHA, and (c) ALE (dark line, floc sludge; light line, biofilm); deconvolution results of CGP in the amide I band under (d) NaAc, (e) GLC, and (f) MeOH as carbon source feeding conditions.

Fig.3 Fluorescence properties of CGP. Fluorescence micrograph of CGP from activated sludge (a), 3D-EEM fluorescence spectra of CGP from Anabaena sp. FACHB-110 (b), floc sludge (c–e) and biofilm (f–h) fed with NaAc, GLC, and MeOH, respectively.

Fig.4 Distribution of synthesis and degradation genes in floc sludge and biofilm under different carbon source feeding conditions. (a) CGP, (b) PHA, and (c) ALE.

Fig.5 (a) Correlation analysis of CGP, PHA and ALE extraction with α-diversity of microbial community, (b) phylum level (Top 15), and (c) genus level (Top 15).

1	K Adames , K Euting , A Broker , A Steinbuchel . (2013). Investigations on three genes in Ralstonia eutropha H16 encoding putative cyanophycin metabolizing enzymes. Applied Microbiology and Biotechnology, 97(8): 3579–3591 https://doi.org/10.1007/s00253-012-4599-6
2	C Amorim de Carvalho, A Ferreira Dos Santos, T J Tavares Ferreira, V N Sousa Aguiar Lira, A R Mendes Barros, A Bezerra Dos Santos (2021). Resource recovery in aerobic granular sludge systems: Is it feasible or still a long way to go? Chemosphere, 274: 129881 https://doi.org/10.1016/j.chemosphere.2021.129881
3	H Chen , Z Chen , X Chu , Y Deng , S Qing , C Sun , Q Wang , H Zhou , H Cheng , W Zhan , Y Wang . (2022a). Temperature mediated the balance between stochastic and deterministic processes and reoccurrence of microbial community during treating aniline wastewater. Water Research, 221: 118741 https://doi.org/10.1016/j.watres.2022.118741
4	X Chen , Y J Lee , T Yuan , Z Lei , Y Adachi , Z Zhang , Y Lin , M C M Van Loosdrecht . (2022b). A review on recovery of extracellular biopolymers from flocculent and granular activated sludges: cognition, key influencing factors, applications, and challenges. Bioresource Technology, 363: 127854 https://doi.org/10.1016/j.biortech.2022.127854
5	Y D Chen , Z Yang , N Q Ren , S H Ho . (2020). Optimizing the production of short and medium chain fatty acids (SCFAs and MCFAs) from waste activated sludge using different alkyl polyglucose surfactants, through bacterial metabolic analysis. Journal of Hazardous Materials, 384: 121384 https://doi.org/10.1016/j.jhazmat.2019.121384
6	M K D Dueholm , M Besteman , E J Zeuner , M Riisgaard-Jensen , M E Nielsen , S Z Vestergaard , S Heidelbach , N S Bekker , P H Nielsen . (2023). Genetic potential for exopolysaccharide synthesis in activated sludge bacteria uncovered by genome-resolved metagenomics. Water Research, 229: 119485 https://doi.org/10.1016/j.watres.2022.119485
7	M Frommeyer , L Wiefel , A Steinbuchel . (2016). Features of the biotechnologically relevant polyamide family “cyanophycins” and their biosynthesis in prokaryotes and eukaryotes. Critical Reviews in Biotechnology, 36(1): 153–164 https://doi.org/10.3109/07388551.2014.946467
8	A González-Martínez , K Calderon , J Gonzalez-Lopez . (2016). New concepts of microbial treatment processes for the nitrogen removal: effect of protein and amino acids degradation. Amino Acids, 48(5): 1123–1130 https://doi.org/10.1007/s00726-016-2185-4
9	X Hao , D Wu , J Li , R Liu , M Van Loosdrecht . (2022). Making waves: a sea change in treating wastewater―Why thermodynamics supports resource recovery and recycling. Water Research, 218: 118516 https://doi.org/10.1016/j.watres.2022.118516
10	R Huang , J Xu , L Xie , H Wang , X Ni . (2022). Energy neutrality potential of wastewater treatment plants: a novel evaluation framework integrating energy efficiency and recovery. Frontiers of Environmental Science & Engineering, 16(9): 117 https://doi.org/10.1007/s11783-022-1549-0
11	C Khomlaem , H Aloui , W G Oh , B S Kim . (2021). High cell density culture of Paracoccus sp. LL1 in membrane bioreactor for enhanced co-production of polyhydroxyalkanoates and astaxanthin. International Journal of Biological Macromolecules, 192: 289–297 https://doi.org/10.1016/j.ijbiomac.2021.09.180
12	N K Kim , N Mao , R Lin , D Bhattacharyya , M C M Van Loosdrecht , Y Lin . (2020). Flame retardant property of flax fabrics coated by extracellular polymeric substances recovered from both activated sludge and aerobic granular sludge. Water Research, 170: 115344 https://doi.org/10.1016/j.watres.2019.115344
13	H Li , J Cheng , H Deng , E Zhao , B Shen , J W Y Lam , K S Wong , H Wu , B S Li , B Z Tang . (2015). Aggregation-induced chirality, circularly polarized luminescence, and helical self-assembly of a leucine-containing AIE luminogen. Journal of Materials Chemistry. C, Materials for Optical and Electronic Devices, 3(10): 2399–2404 https://doi.org/10.1039/C4TC02726D
14	H Li , J Zhang , L Shen , Z Chen , Y Zhang , C Zhang , Q Li , Y Wang . (2019). Production of polyhydroxyalkanoates by activated sludge: correlation with extracellular polymeric substances and characteristics of activated sludge. Chemical Engineering Journal, 361: 219–226 https://doi.org/10.1016/j.cej.2018.12.066
15	J Li , X Hao , W Gan , M C M Van Loosdrecht , Y Wu . (2021a). Recovery of extracellular biopolymers from conventional activated sludge: potential, characteristics and limitation. Water Research, 205: 117706 https://doi.org/10.1016/j.watres.2021.117706
16	J Li , X Hao , W C M Gan , M Van Loosdrecht , Y Wu . (2022). Controlling factors and involved mechanisms on forming alginate like extracellular polymers in flocculent sludge. Chemical Engineering Journal, 439: 135792 https://doi.org/10.1016/j.cej.2022.135792
17	Y Y Li , X W Huang , X Y Li . (2021b). Use of a packed-bed biofilm reactor to achieve rapid formation of anammox biofilms for high-rate nitrogen removal. Journal of Cleaner Production, 321: 128999 https://doi.org/10.1016/j.jclepro.2021.128999
18	L Lu , J S Guest , C A Peters , X Zhu , G H Rau , Z J Ren . (2018). Wastewater treatment for carbon capture and utilization. Nature Sustainability, 1(12): 750–758 https://doi.org/10.1038/s41893-018-0187-9
19	Z Lu , J Ye , Z Chen , L Xiao , L Lei , B P Han , H W Paerl . (2022). Cyanophycin accumulated under nitrogen-fluctuating and high-nitrogen conditions facilitates the persistent dominance and blooms of Raphidiopsis raciborskii in tropical waters. Water Research, 214: 118215 https://doi.org/10.1016/j.watres.2022.118215
20	T Miyakawa , J Yang , M Kawasaki , N Adachi , A Fujii , Y Miyauchi , T Muramatsu , T Moriya , T Senda , M Tanokura . (2022). Structural bases for aspartate recognition and polymerization efficiency of cyanobacterial cyanophycin synthetase. Nature Communications, 13(1): 5097 https://doi.org/10.1038/s41467-022-32834-8
21	P Qin , H Cui , P Li , S Wang , S Fan , J Lu , M Sun , H Zhang , S Wang , X Su . et al.. (2023). Early stage of biofilm assembly on microplastics is structured by substrate size and bacterial motility. iMeta, 2023: e121 https://doi.org/10.1002/imt2.121
22	U Remminghorst , B H Rehm . (2006). In vitro alginate polymerization and the functional role of alg8 in alginate production by Pseudomonas aeruginosa. Applied and Environmental Microbiology, 72(1): 298–305 https://doi.org/10.1128/AEM.72.1.298-305.2006
23	I Sharon , A S Haque , M Grogg , I Lahiri , D Seebach , A E Leschziner , D Hilvert , T M Schmeing . (2021). Structures and function of the amino acid polymerase cyanophycin synthetase. Nature Chemical Biology, 17(10): 1101–1110 https://doi.org/10.1038/s41589-021-00854-y
24	I Sharon , S Pinus , M Grogg , N Moitessier , D Hilvert , T M Schmeing . (2022). A cryptic third active site in cyanophycin synthetase creates primers for polymerization. Nature Communications, 13(1): 3923 https://doi.org/10.1038/s41467-022-31542-7
25	R D Simon , N H Lawry , G L Mclendon . (1980). Structural characterization of the cyanophycin granule polypeptide of Anabaena cylindrica by circular dichroism and raman spectroscopy. Biochimica et Biophysica Acta. Protein Structure, 626(2): 277–281 https://doi.org/10.1016/0005-2795(80)90121-X
26	C O Tuck , E Perez , I T Horvath , R A Sheldon , M Poliakoff . (2012). Valorization of biomass: deriving more value from waste. Science, 337(6095): 695–699 https://doi.org/10.1126/science.1218930
27	M C van Loosdrecht , D Brdjanovic . (2014). Anticipating the next century of wastewater treatment. Science, 344(6191): 1452–1453 https://doi.org/10.1126/science.1255183
28	L Wang , D H Wang , G H Zhang , D Xu , W X Deng . (2016). Crystal structure, spectroscopic investigation and thermal properties of l-lysine p-toluenesulfonate. Journal of Molecular Structure, 1108: 179–186 https://doi.org/10.1016/j.molstruc.2015.11.019
29	W Wu , B Liu . (2021). Aggregation-induced emission: challenges and opportunities. National Science Review, 8(6): nwaa222 https://doi.org/10.1093/nsr/nwaa222
30	C Zhang, P Hu, Q Liu, Z Lu, B Cao, Y Tang, T Hao (2022). Biopolymer recovery from waste activated sludge toward self-healing mortar crack. Science of the Total Environment, 858(Pt 3): 160107 https://doi.org/10.1016/j.scitotenv.2022.160107
31	X Zhang , Y Lin , Q Wu , Y Wang , G Q Chen . (2020). Synthetic biology and genome-editing tools for improving PHA metabolic engineering. Trends in Biotechnology, 38(7): 689–700 https://doi.org/10.1016/j.tibtech.2019.10.006
32	Z Zhang , Y Zhou , J Zhang , S Xia , S W Hermanowicz . (2016). Effects of short-time aerobic digestion on extracellular polymeric substances and sludge features of waste activated sludge. Chemical Engineering Journal, 299: 177–183 https://doi.org/10.1016/j.cej.2016.04.047
33	J Zhu , J Wang , Y P Chen , T P Qing , P Zhang , B Feng . (2022). Quantitative proteomics and phosphoproteomics elucidate the molecular mechanism of nanostructured TiO2-stimulated biofilm formation. Journal of Hazardous Materials, 432: 128709 https://doi.org/10.1016/j.jhazmat.2022.128709
34	K Ziegler , R Deutzmann , W Lockau . (2002). Cyanophycin synthetase-like enzymes of non-cyanobacterial eubacteria: characterization of the polymer produced by a recombinant synthetase of Desulfitobacterium hafniense. Journal of Biosciences, 57(5–6): 522–529 https://doi.org/10.1515/znc-2002-5-621
35	K Zou , Y Huang , B Feng , T P Qing , P Zhang , Y P Chen . (2022). Cyanophycin granule polypeptide: a neglected high value-added biopolymer, synthesized in activated sludge on a large scale. Applied and Environmental Microbiology, 88(14): e0074222 https://doi.org/10.1128/aem.00742-22

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