Please wait a minute...
Frontiers of Environmental Science & Engineering

ISSN 2095-2201

ISSN 2095-221X(Online)

CN 10-1013/X

Postal Subscription Code 80-973

2018 Impact Factor: 3.883

Front. Environ. Sci. Eng.    2017, Vol. 11 Issue (1) : 5    https://doi.org/10.1007/s11783-017-0896-8
RESEARCH ARTICLE |
Investigation of polyhydroxyalkanoates (PHAs) biosynthesis from mixed culture enriched by valerate-dominant hydrolysate
Jiuxiao Hao,Xiujin Wang,Hui Wang()
State Key Joint Laboratory of Environment Simulation and Pollution Control, School of Environment, Tsinghua University, Beijing 100084, China
 Download: PDF(411 KB)   HTML
 Export: BibTeX | EndNote | Reference Manager | ProCite | RefWorks
Abstract

Valerate-hydrolysate enriched culture showed great ability to produce 3HV and 3H2MV.

Valerate-hydrolysate enriched culture had more Brevundimonas in the community.

Mixed iso-/n-valerate was effective at balancing microbial growth and PHAs synthesis.

Co-substrates of valerate and propionate raised the fraction of 3HV and 3H2MV.

The production of polyhydroxyalkanoates (PHAs) with a high fraction of 3-hydroxyvalerate (3HV) and 3-hydroxy-2-methylvalerate (3H2MV) from mixed culture enriched by valerate-dominant hydrolysate was evaluated in this study. After long-term enrichment, the culture showed strong ability to synthesize 3HV and 3H2MV, even with acetate-dominant substrate. The ultilization of single or mixed iso-/n-valerate by the enriched culture showed that the mixture of iso-valerate and n-valerate was more efficient substrate than any single in terms of balancing microbial growth and PHAs synthesis. Besides, through comparing the kinetics and stoichiometry of the tests supplying valerate and propionate, the enriched culture with equivalent valerate and propionate (1:1 molar ratio) exhibited superior PHAs production performances to pure valerate or propionate, attaining more than 70 mol% of 3HV and 3H2MV. The above findings reveal that valerate-dominant hydrolysate is a kind of suitable substrate to enrich PHAs producing culture with great capability to synthesize 3HV and 3H2MV monomers, thus improving product properties than pure poly(3-hydroxybutyrate) (P3HB); also 3HV and 3H2MV production behaviors can be regulated by the type of odd-carbon VFAs in the substrate.

Keywords Polyhydroxyalkanoates (PHAs)      Valerate      Mixed culture      3-hydroxyvalerate (3HV)      Propionate     
PACS:     
Fund: 
Corresponding Authors: Hui Wang   
Issue Date: 23 December 2016
 Cite this article:   
Jiuxiao Hao,Xiujin Wang,Hui Wang. Investigation of polyhydroxyalkanoates (PHAs) biosynthesis from mixed culture enriched by valerate-dominant hydrolysate[J]. Front. Environ. Sci. Eng., 2017, 11(1): 5.
 URL:  
http://academic.hep.com.cn/fese/EN/10.1007/s11783-017-0896-8
http://academic.hep.com.cn/fese/EN/Y2017/V11/I1/5
VFAs/(C mmol·L1) biomass enrichment batch tests a)
valerate-dominant hydrolysate
(#1)
acetate-dominant hydrolysate b)
(#2)
single iso-Val
(#3)
single n-Val
(#4)
mixed iso-/n-Val c)
(#5)
pure Val c)
(#5)
pure Pro
(#6)
Val-Pro 1:1
(#7)
total 100 100 100 100 100 100 100 100
acetate 25 50 0 0 0 0 0 0
propionate 12.5 25 0 0 0 0 100 50
butyrate 12.5 25 0 0 0 0 0 0
iso-valerate 25 0 100 0 50 50 0 25
n-valerate 25 0 0 100 50 50 0 25
Tab.1  Composition of VFAs in different substrates for PHAs culture enrichment and batch experiments
Fig.1  Cyclic profiles of (a) substrate utilization and PHAs production in two SBRs, and the usage of individual VFA in (b) valerate-hydrolysate SBR and (c) acetate-hydrolysate SBR (“Valerate” includes iso-valerate and n-valerate, bars= S.D., n = 3)
Fig.2  Concentrations of CDW, PHAs and active biomass (X) during one cycle in (a) valerate-hydrolysate SBR and (b) acetate-hydrolysate SBR, and (c) compositions of synthesized PHAs (The red and blue dashed lines mark the end of feast phase, bars= S.D., n = 3)
parameter valerate-hydrolysate
enriched culture
acetate-hydrolysate
enriched culture
species richness species richness
Phylum level a) Proteobacteria 85.25% Proteobacteria 71.00%
Bacteroidetes 14.51% Bacteroidetes 28.64%
Genus level a) Brevundimonas 80.90% Brevundimonas 63.81%
Sphingobacterium 14.05% Sphingobacterium 27.78%
Paenochrobactrum 1.42%
Devosia 1.31%
Pseudaminobacter 1.03%
Chao 1 richness b) 297.38 384.77
ACE richness b) 334.82 399.15
Tab.2  Relative taxonomic abundance at phylum and genus levels for valerate-hydrolysate enriched culture and acetate-hydrolysate enriched culture, and their species richness
Fig.3  (a) Substrate utilization, (b) PHAs production and (c) PHAs composition from batch test by valerate-hydrolysate enriched culture with acetate-hydrolysate (bars= S.D., n = 3)
Fig.4  PHAs production behaviors of valerate-hydrolysate enriched culture with (a) iso-valerate, (b) n-valerate, and (c) mixed iso-/n-valerate, as well as (d) their PHAs compositions at the moment with maximum PHAs concentration (bars= S.D., n = 3)
substrate b) qValc) /(C mmol·(L·h)1) qProc) /(C mmol·(L·h)1) PHAmax /(g·L1) Xmax /(g·L1) YPHA/Sd) /(C mmol per C mmol) YX/Sd) /(C mmol per C mmol) 3HB/3HV/3H2MV /mol% qPHAd) /(g·(L·h)1) q3HV24h /(g·(L·h)1) q3H2MV24h /(g·(L·h)1)
24h 36h
pure valerate 1.63 30 h: 0.54 48 h: 0.82 0.45 0.3 65.21/25.84/8.95 53.26/35.10/11.64 0.018 0.0058 0.0022
pure propionate 2.27 18 h: 0.36 18 h: 1.48 0.38 1.45 66.31/19.29/14.40 62.84/18.13/19.03 0.02 0.0026 0.0021
valerate-propionate 1:1 1.41 1.32 30 h: 0.95 12 h: 0.95 0.48 0.74 28.42/67.27/9.31 27.15/68.51/4.34 0.032 0.0246 0.0038
Tab.3  Kinetics and stoichiometry of batch PHAs production tests from valerate-hydrolysate enriched culture with valerate and propionate as substrate a)
1 Anderson A J, Dawes E A. Occurrence, metabolism, metabolic role, and industrial uses of bacterial polyhydroxyalkanoates. Microbiological Reviews, 1990, 54(4): 450–472
pmid: 2087222
2 Kleerebezem R, van Loosdrecht M C M. Mixed culture biotechnology for bioenergy production. Current Opinion in Biotechnology, 2007, 18(3): 207–212
https://doi.org/10.1016/j.copbio.2007.05.001 pmid: 17509864
3 Johnson K, Jiang Y, Kleerebezem R, Muyzer G, van Loosdrecht M C M. Enrichment of a mixed bacterial culture with a high polyhydroxyalkanoate storage capacity. Biomacromolecules, 2009, 10(4): 670–676
https://doi.org/10.1021/bm8013796 pmid: 19193058
4 Hanson A J, Guho N M, Paszczynski A J, Coats E R. Community proteomics provides functional insight into polyhydroxyalkanoate production by a mixed microbial culture cultivated on fermented dairy manure. Applied Microbiology and Biotechnology, 2016, 100(18): 7957–7976
https://doi.org/10.1007/s00253-016-7576-7 pmid: 27147532
5 Jiang Y, Chen Y, Zheng X. Efficient polyhydroxyalkanoates production from a waste-activated sludge alkaline fermentation liquid by activated sludge submitted to the aerobic feeding and discharge process. Environmental Science & Technology, 2009, 43(20): 7734–7741
https://doi.org/10.1021/es9014458 pmid: 19921887
6 Sudesh K, Abe H, Doi Y. Synthesis, structure and properties of polyhydroxyalkanoates: biological polyesters. Progress in Polymer Science, 2000, 25(10): 1503–1555
https://doi.org/10.1016/S0079-6700(00)00035-6
7 Rehm B H, Steinbüchel A. Biochemical and genetic analysis of PHA synthases and other proteins required for PHA synthesis. International Journal of Biological Macromolecules, 1999, 25(1–3): 3–19
https://doi.org/10.1016/S0141-8130(99)00010-0 pmid: 10416645
8 Arcos-Hernández M V, Laycock B, Donose B C, Pratt S, Halley P, Al-Luaibi S, Werker A, Lant P A. Physicochemical and mechanical properties of mixed culture polyhydroxyalkanoate (PHBV). European Polymer Journal, 2013, 49(4): 904–913
https://doi.org/10.1016/j.eurpolymj.2012.10.025
9 Slater S, Houmiel K L, Tran M, Mitsky T A, Taylor N B, Padgette S R, Gruys K J. Multiple b-ketothiolases mediate poly(b-hydroxyalkanoate) copolymer synthesis in Ralstonia eutropha. Journal of Bacteriology, 1998, 180(8): 1979–1987
pmid: 9555876
10 Steinbüchel A, Lütke-Eversloh T. Metabolic engineering and pathway construction for biotechnological production of relevant polyhydroxyalkanoates in microorganisms. Biochemical Engineering Journal, 2003, 16(2): 81–96
https://doi.org/10.1016/S1369-703X(03)00036-6
11 Albuquerque M G, Martino V, Pollet E, Avérous L, Reis M A. Mixed culture polyhydroxyalkanoate (PHA) production from volatile fatty acid (VFA)-rich streams: effect of substrate composition and feeding regime on PHA productivity, composition and properties. Journal of Biotechnology, 2011, 151(1): 66–76
https://doi.org/10.1016/j.jbiotec.2010.10.070 pmid: 21034785
12 Gobi K, Vadivelu V M. Dynamics of polyhydroxyalkanoate accumulation in aerobic granules during the growth-disintegration cycle. Bioresource Technology, 2015, 196: 731–735
https://doi.org/10.1016/j.biortech.2015.07.083 pmid: 26235884
13 Khanna S, Srivastava A K. Production of poly(3-hydroxybutyric-co-3-hydroxyvaleric acid) having a high hydroxyvalerate content with valeric acid feeding. Journal of Industrial Microbiology & Biotechnology, 2007, 34(6): 457–461
https://doi.org/10.1007/s10295-007-0207-7 pmid: 17268758
14 Chang H F, Chang W C, Tsai C Y. Synthesis of poly(3-hydroxybutyrate/3-hydroxyvalerate) from propionate-fed activated sludge under various carbon sources. Bioresource Technology, 2012, 113: 51–57
https://doi.org/10.1016/j.biortech.2011.12.138 pmid: 22277212
15 Lemos P C, Serafim L S, Reis M A. Synthesis of polyhydroxyalkanoates from different short-chain fatty acids by mixed cultures submitted to aerobic dynamic feeding. Journal of Biotechnology, 2006, 122(2): 226–238
https://doi.org/10.1016/j.jbiotec.2005.09.006 pmid: 16253370
16 Albuquerque M G, Torres C A, Reis M A. Polyhydroxyalkanoate (PHA) production by a mixed microbial culture using sugar molasses: effect of the influent substrate concentration on culture selection. Water Research, 2010, 44(11): 3419–3433
https://doi.org/10.1016/j.watres.2010.03.021 pmid: 20427069
17 Bengtsson S, Werker A, Christensson M, Welander T. Production of polyhydroxyalkanoates by activated sludge treating a paper mill wastewater. Bioresource Technology, 2008, 99(3): 509–516
https://doi.org/10.1016/j.biortech.2007.01.020 pmid: 17360180
18 Rajagopal R, Béline F. Anaerobic hydrolysis and acidification of organic substrates: determination of anaerobic hydrolytic potential. Bioresource Technology, 2011, 102(10): 5653–5658
https://doi.org/10.1016/j.biortech.2011.02.068 pmid: 21444204
19 Zhou A, Guo Z, Yang C, Kong F, Liu W, Wang A. Volatile fatty acids productivity by anaerobic co-digesting waste activated sludge and corn straw: effect of feedstock proportion. Journal of Biotechnology, 2013, 168(2): 234–239
https://doi.org/10.1016/j.jbiotec.2013.05.015 pmid: 23751505
20 Hao J, Wang H. Volatile fatty acids productions by mesophilic and thermophilic sludge fermentation: biological responses to fermentation temperature. Bioresource Technology, 2015, 175: 367–373
https://doi.org/10.1016/j.biortech.2014.10.106 pmid: 25459844
21 Xiong H, Chen J, Wang H, Shi H. Influences of volatile solid concentration, temperature and solid retention time for the hydrolysis of waste activated sludge to recover volatile fatty acids. Bioresource Technology, 2012, 119: 285–292
https://doi.org/10.1016/j.biortech.2012.05.126 pmid: 22750494
22 Jia Q, Wang H, Wang X. Dynamic synthesis of polyhydroxyalkanoates by bacterial consortium from simulated excess sludge fermentation liquid. Bioresource Technology, 2013, 140: 328–336
https://doi.org/10.1016/j.biortech.2013.04.105 pmid: 23711941
23 Doi Y, Kunioka M, Nakamura Y, Soga K. Nuclear magnetic resonance studies on poly (b-hydroxybutyrate) and a copolyester of b-hydroxybutyrate and b-hydroxyvalerate isolated from Alcaligenes eutrophus H16. Macromolecules, 1986, 19(11): 2860–2864
https://doi.org/10.1021/ma00165a033
24 Feng L, Chen Y, Zheng X. Enhancement of waste activated sludge protein conversion and volatile fatty acids accumulation during waste activated sludge anaerobic fermentation by carbohydrate substrate addition: the effect of pH. Environmental Science & Technology, 2009, 43(12): 4373–4380
https://doi.org/10.1021/es8037142 pmid: 19603649
25 Bhuwal A K, Singh G, Aggarwal N K, Goyal V, Yadav A. Isolation and screening of polyhydroxyalkanoates producing bacteria from pulp, paper, and cardboard industry wastes. International Journal of Biomaterials, 2013, 2013
26 Silva J A, Tobella L M, Becerra J, Godoy F, Martínez M A. Biosynthesis of poly-b-hydroxyalkanoate by Brevundimonas vesicularis LMG P-23615 and Sphingopyxis macrogoltabida LMG 17324 using acid-hydrolyzed sawdust as carbon source. Journal of Bioscience and Bioengineering, 2007, 103(6): 542–546
https://doi.org/10.1263/jbb.103.542 pmid: 17630126
27 Terrill T, Douglas G, Foote A, Purchas R, Wilson G, Barry T. Effect of condensed tannins upon body growth, wool growth and rumen metabolism in sheep grazing sulla (Hedysarum coronarium) and perennial pasture. Journal of Agricultural Science, 1992, 119(2): 265–273
https://doi.org/10.1017/S0021859600014192
28 Hilliou L, Teixeira P F, Machado D, Covas J A, Oliveira C S, Duque A F, Reis M A. Effects of fermentation residues on the melt processability and thermomechanical degradation of PHBV produced from cheese whey using mixed microbial cultures. Polymer Degradation & Stability, 2016, 128: 269–277
https://doi.org/10.1016/j.polymdegradstab.2016.03.031
29 Ucisik A S, Henze M. Biological hydrolysis and acidification of sludge under anaerobic conditions: the effect of sludge type and origin on the production and composition of volatile fatty acids. Water Research, 2008, 42(14): 3729–3738
https://doi.org/10.1016/j.watres.2008.06.010 pmid: 18703214
30 Gou M, Zeng J, Wang H Z, Tang Y Q, Shigematsu T, Morimura S, Kida K. Microbial community structure and dynamics of starch-fed and glucose-fed chemostats during two years of continuous operation. Frontiers of Environmental Science & Engineering, 2016, 10(2): 368–380
https://doi.org/10.1007/s11783-015-0815-9
31 Steinbüchel A, Valentin H E. Diversity of bacterial polyhydroxyalkanoic acids. FEMS Microbiology Letters, 1995, 128(3): 219–228
https://doi.org/10.1016/0378-1097(95)00125-O
32 Michinaka A, Arou J, Onuki M, Satoh H, Mino T. Analysis of polyhydroxyalkanoate (PHA) synthase gene in activated sludge that produces PHA containing 3-hydroxy-2-methylvalerate. Biotechnology and Bioengineering, 2007, 96(5): 871–880
https://doi.org/10.1002/bit.21085 pmid: 16933327
33 Oehmen A, Keller-Lehmann B, Zeng R J, Yuan Z, Keller J. Optimisation of poly-b-hydroxyalkanoate analysis using gas chromatography for enhanced biological phosphorus removal systems. Journal of Chromatography. A, 2005, 1070(1–2): 131–136
https://doi.org/10.1016/j.chroma.2005.02.020 pmid: 15861796
34 Chen Z, Guo Z, Wen Q, Huang L, Bakke R, Du M. A new method for polyhydroxyalkanoate (PHA) accumulating bacteria selection under physical selective pressure. International Journal of Biological Macromolecules, 2015, 72: 1329–1334
https://doi.org/10.1016/j.ijbiomac.2014.10.027 pmid: 25450834
35 Liu C, Li J Z, Wang S, Nies L. A syntrophic propionate-oxidizing microflora and its bioaugmentation on anaerobic wastewater treatment for enhancing methane production and COD removal. Frontiers of Environmental Science & Engineering, 2016, 10(4): 19
https://doi.org/10.1007/s11783-016-0856-8
[1] FSE-16059-OF-HJX_suppl_1 Download
[1] Kishore Gopalakrishnan, Javad Roostaei, Yongli Zhang. Mixed culture of Chlorella sp. and wastewater wild algae for enhanced biomass and lipid accumulation in artificial wastewater medium[J]. Front. Environ. Sci. Eng., 2018, 12(4): 14-.
Viewed
Full text


Abstract

Cited

  Shared   
  Discussed