Investigation of polyhydroxyalkanoates (PHAs) biosynthesis from mixed culture enriched by valerate-dominant hydrolysate

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.

Graphical abstract

Keywords

Polyhydroxyalkanoates (PHAs) / Valerate / Mixed culture / 3-hydroxyvalerate (3HV) / Propionate

References

[1] Anderson A J, Dawes E A. Occurrence, metabolism, metabolic role, and industrial uses of bacterial polyhydroxyalkanoates. Microbiological Reviews, 1990, 54(4): 450–472

[2] Kleerebezem R, van Loosdrecht M C M. Mixed culture biotechnology for bioenergy production. Current Opinion in Biotechnology, 2007, 18(3): 207–212

[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

[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

[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

[6] Sudesh K, Abe H, Doi Y. Synthesis, structure and properties of polyhydroxyalkanoates: biological polyesters. Progress in Polymer Science, 2000, 25(10): 1503–1555

[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

[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

[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

[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

[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

[12] Gobi K, Vadivelu V M. Dynamics of polyhydroxyalkanoate accumulation in aerobic granules during the growth-disintegration cycle. Bioresource Technology, 2015, 196: 731–735

[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

[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

[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

[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

[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

[18] Rajagopal R, Béline F. Anaerobic hydrolysis and acidification of organic substrates: determination of anaerobic hydrolytic potential. Bioresource Technology, 2011, 102(10): 5653–5658

[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

[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

[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

[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

[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

[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

[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

[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

[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

[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

[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

[31] Steinbüchel A, Valentin H E. Diversity of bacterial polyhydroxyalkanoic acids. FEMS Microbiology Letters, 1995, 128(3): 219–228

[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

[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

[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

[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

Acknowledgements

The authors would like to thank the funding of National Key Research on Water Environment Pollution Control in China (No. 2012ZX07313-001).

The authors declare that they have no conflict of interest.

Electronic Supplementary Material

Supplementary material is available in the online version of this article at http://dx.doi.org/10.1007/s11783-017-0896-8 and is accessible for authorized users.

Funding

RIGHTS & PERMISSIONS

2017 Higher Education Press and Springer-Verlag Berlin Heidelberg