Purpose of Review
This review paper focuses on the recent advances made in the area of polyhydroxyalkanoate (PHA) production for their commercialization purposes. Based on recent literature reports, the paper summarizes the major challenges faced by researchers concerning the industrial aspects of bioprocessing and downstream recovery, and the development of superior strains via metabolic engineering approaches for enhanced PHA production.
Previous reports have shown that the researchers are now shifting towards upcycling of plastic waste and recycling of organic waste for both bioenergy and bio-polyester production, next-generation industrial biotechnology techniques, and metabolic engineering strategies of ribosome-binding site optimization, CRISPR/Cas9, and engineering cell growth pattern and shapes for downstream recovery, for high-throughput and cost-efficient PHA production at industrial scale.
The recent findings indicate that the use of organic waste substrates, development of high-yielding microbial strains, and multistage cultivation strategies with cell recycling could help in minimizing the production costs of PHA and facilitate their commercialization. Nowadays, the continuous and non-sterile bioreactor processes based on mixed cultures or extremophiles, and/or mixed substrates, along with environmentally benign low-cost downstream recovery strategies are recommended. Therefore, the trend is towards the application of combined technologies to maximize PHA production while reducing production costs.
This is a preview of subscription content, log in to check access.
Buy single article
Instant access to the full article PDF.
Price includes VAT for USA
Subscribe to journal
Immediate online access to all issues from 2019. Subscription will auto renew annually.
This is the net price. Taxes to be calculated in checkout.
Papers of particular interest, published recently, have been highlighted as: •• Of major importance
Vijay R, Tarika K. Production of polyhydroxyalkanoates (PHAs) using synthetic biology and metabolic engineering approaches. Res J Biotechnol. 2018;13(1):99–109.
•• Pérez-Rivero C, Lopez-Gomez JP, Roy I. A sustainable approach for the downstream processing of bacterial polyhydroxyalkanoates: state-of-the-art and latest developments. Biochem Eng J. 2019;150:107283 This review paper provides an overview of all the recent approaches involved in downstream recovery of PHA. This paper specially focuses on the green technologies for PHA recovery with scale-up possibilities. The strengths and weaknesses of all the strategies have been discussed.
Rodriguez-Perez S, Serrano A, Pantión AA, Alonso-Fariñas B. Challenges of scaling-up PHA production from waste streams. A review. J Environ Manage. 2018;205:215–30.
Pagliano G, Ventorino V, Panico A, Pepe O. Integrated systems for biopolymers and bioenergy production from organic waste and by-products: a review of microbial processes. Biotechnol Biofuels. 2017;10(1):113.
Gahlawat G. Polyhydroxyalkanoates biopolymers: production strategies. In: Springer briefs in molecular science: biobased polymers: Springer International Publishing; 2019.
Gahlawat G, Soni KS. Study on sustainable recovery and extraction of polyhydroxyalkanoates (PHAs) produced by Cupriavidus necator using waste glycerol for medical applications. Chem Biochem Eng Q. 2019;33:99–110.
Ong SY, Idris ZL, Pyary S, Sudesh K. A novel biological recovery approach for PHA employing selective digestion of bacterial biomass in animals. Appl Microbiol Biotechnol. 2018;102:2117–27.
Zhang X, Lin Y, Wu Q, Wang Y, Chen GQ. Synthetic biology and genome-editing tools for improving PHA metabolic engineering. Trends Biotechnol. 2019;38:689–700. https://doi.org/10.1016/j.tibtech.2019.10.006.
Elhadi D, Lv L, Jiang XR, Wu H, Chen GQ. CRISPRi engineering E. coli for morphology diversification. Metab Eng. 2016;38:358–69.
Ye J, Hu D, Che X, Jiang X, Li T, Chen J, et al. Engineering of Halomonas bluephagenesis for low cost production of poly (3-hydroxybutyrate-co-4-hydroxybutyrate) from glucose. Metab Eng. 2018;47:143–52.
•• Koller M, Braunegg G. Advanced approaches to produce polyhydroxyalkanoate (PHA) biopolyesters in a sustainable and economic fashion. EuroBiotech J. 2018;2:89–103 The writer has given the elaborated overview of second-generation carbon substrates for the bioplastic production, which is generated from industrial waste and provides sustainable means for acquiring raw materials.
Pan C, Tan GY, Ge L, Chen CL, Wang JY. Two-stage microbial conversion of crude glycerol to 1,3-propanediol and polyhydroxyalkanoates after pretreatment. J Environ Mange. 2019;232:615–24.
Amini M, Yousefi-Massumabad H, Younesi H, Abyar H, Bahramifar N. Production of the polyhydroxyalkanoate biopolymer by Cupriavidus necator using beer brewery wastewater containing maltose as a primary carbon source. J Environ Chem Eng. 2019;8(1):103588.
Heepkaew P, Suwannasilp BB. Polyhydroxyalkanoate production using two-stage continuous stirred tank activated sludge systems with glycerol as a carbon source. J Chem Technol Biotechnol. 2019. https://doi.org/10.1002/jctb.6304.
Joyline M, Aruna K. Production and characterization of polyhydroxyalkanoates (PHA) by Bacillus megaterium strain JHA using inexpensive agro-industrial wastes. Int J Rec Sci Res. 2019;10(7):33359–74.
Dañez JCA, Requiso PJ, Alfafara CG, Nayve FRP Jr, Ventura JRS. Optimization of fermentation factors for polyhydroxybutyrate (PHB) production using Bacillus megaterium PNCM 1890 in simulated glucose-xylose hydrolysates from agricultural residues. Phili. J. Sci. 2020;149:163–75.
Obruca S, Benesova P, Marsalek L, Marova I. Use of lignocellulosic materials for PHA production. Chem Biochem Eng. 2015;29:135–44.
Johnston B, Radecka I, Hill D, Chiellini E, Ilieva V, Sikorska W, et al. The microbial production of polyhydroxyalkanoates from waste polystyrene fragments attained using oxidative degradation. Polym. 2018;10:1–22.
López JC, Arnáiz E, Merchán L, Lebrero R, Muñoz R. Biogas-based polyhydroxyalkanoates production by Methylocystis hirsuta : a step further in anaerobic digestion biorefineries. Chem Eng J. 2018;333:529–36.
Zohri NAA, Kamal El-Dean AM, Abuo-Dobara MI, Ali MI, et al. Production of polyhydroxyalkanoate by local strain of Bacillus megaterium AUMC b 272 utilizing sugar beet wastewater and molasses. Egy Sug J. 2019;13:45–70.
Domingos JM, Puccio S, Martinez GA, Amaral N, Reis MA, Bandini M, et al. Cheese whey integrated valorisation: production, concentration and exploitation of carboxylic acids for the production of polyhydroxyalkanoates by a fed-batch culture. Chem Eng J. 2018;336:7–53.
Marques Monteiro Amaro TM, Rosa F, Comi G, Iacumin L. Prospects for the use of whey for polyhydroxyalkanoate (PHA) production. Front Microbiol. 2018;10:1–12. https://doi.org/10.3389/fmicb.2019.00992.
Poblete-Castro I, Wittmann C, Nikel PI. Biochemistry, genetics and biotechnology of glycerol utilization in Pseudomonas sp. Microb Biotechnol. 2019;13(1):32–53.
Garcia-Gonzalez L, De Wever H. Valorization of CO2-rich off-gases to biopolymers through biotechnological process. FEMS Microbio Lett. 2017;364(20):1–8.
Chen GQ, Jiang XR. Next generation industrial biotechnology based on extremophilic bacteria. Curr Opin Biotechnol. 2018;50:94–100.
Zhang X, Lin Y, Chen GQ. Halophiles as chassis for bioproduction. Adv Biosyst. 2018;2:1–12.
Oliveira CS, Silva CE, Carvalho G, Reis MA. Strategies for efficiently selecting PHA producing mixed microbial cultures using complex feedstocks: feast and famine regime and uncoupled carbon and nitrogen availabilities. New Biotechnol. 2016;37:69–79.
Yu L, Wu F, Chen G. Next-generation industrial biotechnology-transforming the current industrial biotechnology into competitive processes. Biotechnol J. 2019;14(9):1800437.
Follonier S, Riesen R, Zinn M. Pilot-scale production of functionalized mcl-PHA from grape pomace 1094 supplemented with fatty acids. Chem Biochem Eng Q. 2015;29:113–21.
Huong KH, Azuraini MJ, Aziz NA, Amirul AAA. Pilot scale production of poly(3-hydroxybutyrate-co-4-hydroxybutyrate) biopolymers with high molecular weight and elastomeric properties. J Biosci Bioeng. 2017;124:76–83.
Werker A, Bengtsson S, Korving L, Hjort M, Anterrieu S, Alexandersson T, et al. Consistent production of high quality PHA using activated sludge harvested from full scale municipal wastewater treatment–PHARIO. Water Sci Technol. 2018;78:2256–69.
Koller M, Vadlja D, Braunegg G, Atlić A, Horvat P. Formal-and high-structured kinetic process modelling and footprint area analysis of binary imaged cells: tools to understand and optimize multistage-continuous PHA biosynthesis. The EuroBiotech J. 2017;1(3):1–9.
Haas C, El-Najjar T, Virgolini N, Smerilli M, Neureiter M. High cell-density production of poly (3-hydroxybutyrate) in a membrane bioreactor. New Biotechnol. 2018;37:117–22.
Chen Z, Huang L, Wen Q, Guo Z. Efficient polyhydroxyalkanoate (PHA) accumulation by a new continuous feeding mode in three-stage mixed microbial culture (MMC) PHA production process. J Biotechnol. 2015;209:68–75.
Vadija D, Koller M, Novak M, Braunegg G, Horvat P. Footprint area analysis of binary imaged Cupriavidus necator cells to study PHB production at balanced, transient, and limited growth conditions in a cascade process. Appl Microbiol Biotechnol. 2016;23:10065–80.
•• Pillai AB, Kumar AJ, Kumarapillai H. Synthetic biology and metabolic engineering approaches for improved production and recovery of bacterial polyhydroxyalkanoates. In: Next generation biomanufacturing technologies: American Chemical Society; 2019. This review addresses the critical aspects of metabolic engineering strategies in polyhydroxyalkanoate production. Recent approaches of synthetic biology are discussed detailing the engineering pathways involved in improvement of PHA production.
Liu MH, Chen YJ, Lee CY. Characterization of medium-chain-length polyhydroxyalkanoate biosynthesis by Pseudomonas mosselii TO7 using crude glycerol. Biosci Biotechnol Biochem. 2018;82(3):532–9.
Kim HS, Oh YH, Jang YA, Kang KH, David Y, Yu JH, et al. Recombinant Ralstonia eutropha engineered to utilize xylose and its use for the production of poly (3-hydroxybutyrate) from sunflower stalk hydrolysate solution. Microb Cell Factories. 2016;15(1):95.
Favaro L, Basaglia M, Casella S. Improving polyhydroxyalkanoate production from inexpensive carbon sources by genetic approaches: a review. Biofuels Bioprod Biorefin. 2019;13(1):208–27.
Yang S, Li S, Jia X. Production of medium chain length polyhydroxyalkanoate from acetate by engineered Pseudomonas putida KT2440. J Ind Microbiol Biotechnol. 2019;46(6):793–800.
Chen J, Li W, Zhang ZZ, Tan TW, Li ZJ. Metabolic engineering of Escherichia coli for the synthesis of polyhydroxyalkanoates using acetate as a main carbon source. Microb Cell Factories. 2018;17(1):102.
Kourmentza C, Plácido J, Venetsaneas N, Burniol-Figols A, Varrone C, Gavala HN, et al. Recent advances and challenges towards sustainable polyhydroxyalkanoate (PHA) production. Bioeng. 2017;4(2):55.
Jiang XR, Wang H, Shen R, Chen GQ. Engineering the bacterial shapes for enhanced inclusion bodies accumulation. Metab Eng. 2015;29:227–37.
Wu H, Chen J, Chen GQ. Engineering the growth pattern and cell morphology for enhanced PHB production by Escherichia coli. Appl Microbiol Biotechnol. 2016a;100(23):9907–16.
Zhang XC, Guo Y, Liu X, Chen XG, Wu Q, Chen GQ. Engineering cell wall synthesis mechanism for enhanced PHB accumulation in E. coli. Metab Eng. 2018;45:32–42.
Wu H, Fan Z, Jiang X, Chen J, Chen GQ. Enhanced production of polyhydroxybutyrate by multiple dividing E. coli. Microb Cell Fact. 2016b;15(1):128.
Ouyang P, Wang H, Hajnal I, Wu Q, Guo Y, Chen GQ. Increasing oxygen availability for improving poly(3-hydroxybutyrate) production by Halomonas. Metab Eng. 2018;45:20–31.
Chen X, Yin J, Ye J, Zhang H, Che X, Ma Y, et al. Engineering Halomonas bluephagenesis TD01 for non-sterile production of poly (3-hydroxybutyrate-co-4-hydroxybutyrate). Bioresour Technol. 2017;244:534–41.
Li M, Chen X, Che X, Zhang H, Wu LP, Du H, et al. Engineering Pseudomonas entomophila for synthesis of copolymers with defined fractions of 3-hydroxybutyrate and medium-chain-length 3-hydroxyalkanoates. Metab Eng. 2019;52:253–62.
Lv L, Ren YL, Chen JC, Wu Q, Chen GQ. Application of CRISPRi for prokaryotic metabolic engineering involving multiple genes, a case study: controllable P (3HB-co-4HB) biosynthesis. Metab Eng. 2015;29:160–8.
Chen Y, Chen XY, Du HT, Zhang X, Ma YM, Chen JC, et al. Chromosome engineering of the TCA cycle in Halomonas bluephagenesis for production of copolymers of 3-hydroxybutyrate and 3-hydroxyvalerate (PHBV). Metab Eng. 2019;54:69–82.
Jung HR, Yang SY, Moon YM, Choi TR, Song HS, Bhatia S, et al. Construction of efficient platform Escherichia coli strains for polyhydroxyalkanoate production by engineering branched pathway. Polym. 2019;11(3):509.
Li D, Lv L, Chen JC, Chen GQ. Controlling microbial PHB synthesis via CRISPRi. Appl Microbiol Biotechnol. 2017;101(14):5861–7.
Shen R, Yin J, Ye JW, Xiang RJ, Ning ZY, Huang WZ, et al. Promoter engineering for enhanced P (3HB-co-4HB) production by Halomonas bluephagenesis. ACS Synth Biol. 2018;7(8):1897–906.
Li T, Li T, Ji W, Wang Q, Zhang H, Chen GQ, et al. Engineering of core promoter regions enables the construction of constitutive and inducible promoters in Halomonas sp. Biotechnol J. 2016;11(2):219–27.
Zhao F, Liu X, Kong A, Zhao Y, Fan X, Ma T, et al. Screening of endogenous strong promoters for enhanced production of medium-chain-length polyhydroxyalkanoates in Pseudomonas mendocina NK-01. Sci Rep. 2019;9(1):1–13.
NAA Z, Ng LM, Foong CP, Tai YT, Nanthini J, Sudesh K. Complete genome sequence of a novel polyhydroxyalkanoate (PHA) producer, Jeongeupia sp. USM3 (JCM 19920) and characterization of its PHA synthases. Curr Microbiol. 2020. https://doi.org/10.1007/s00284-019-01852-z.
Wang Y, Chung A, Chen GQ. Synthesis of medium-chain-length polyhydroxyalkanoate homopolymers, random copolymers, and block copolymers by an engineered strain of Pseudomonas entomophila. Adv Healthc Mater. 2017;6(7):1601017.
Chen GQ, Jiang XR. Engineering bacteria for enhanced polyhydroxyalkanoates (PHA) biosynthesis. Synth Syst Biotechnol. 2017;2(3):192–7.
Meng DC, Chen GQ. Synthetic biology of polyhydroxyalkanoates (PHA). In: Synthetic biology–metabolic engineering. Cham: Springer; 2017. p. 147–74.
Macagnan KL, Alves MI, Moreira AS. Approaches for enhancing extraction of bacterial polyhydroxyalkanoates for industrial applications. In: Kalia VC, editor. Biotechnological applications of polyhydroxyalkanoates. Singapore: Springer Nature; 2019. p. 389–408.
Kumar M, Ghosh P, Khosla K, Thakur IS. Recovery of polyhydroxyalkanoates from municipal secondary wastewater sludge. Bioresour Technol. 2018;255:111–5.
Mohammed S, Panda AN, Ray L. An investigation for recovery of polyhydroxyalkanoates (PHA) from Bacillus sp. BPPI-14 and Bacillus sp. BPPI-19 isolated from plastic waste landfill. Int J Biol Macromol. 2019;134:1085–96.
Mannina G, Presti D, Montiel-Jarillo G, Suárez-Ojeda ME. Bioplastic recovery from wastewater: a new protocol for polyhydroxyalkanoates (PHA) extraction from mixed microbial cultures. Bioresour Technol. 2019;282:361–9.
Leong YK, Show PL, Lan JCW, Loh HS, Yap YJ, Ling TC. Extraction and purification of polyhydroxyalkanoates(PHAs): application of thermoseparating aqueous two-phase extraction. J Polym Res. 2017;24:158–67.
Pillai AB, Kumar AJ, Kumarapillai H. Enhanced production of poly(3-hydroxybutyrate) in recombinant Escherichia coli and EDTA-microwave-assisted cell lysis for polymer recovery. AMB Expr. 2018;8:142–57.
Murugan P, Han L, Gan CY, Maurer FHJ, Sudesh K. A new biological recovery approach for PHA using meal worm, Tenebrio molitor. J Biotechnol. 2016;239:98–105.
Kunasundari B, Arza CR, Maurer FHJ, Murugaiyah V, Kaur G, Sudesh K. Biological recovery and properties of poly(3-hydroxybutyrate) from Cupriavidus necator H16. Sep Purif Technol. 2017;172:1–6.
Gao Y, Feng X, Xian M, Wang Q, Zhao G. Inducible cell lysis systems in microbial production of bio-based chemicals. Appl Microbiol Biotechnol. 2013;97:7121–9.
Alves MI, Macagnan KL, Rodrigues AA, de Assis DA, Torres MM, de Oliveira PD, et al. Poly (3-hydroxybutyrate)-P (3HB): review of production process technology. Ind Biotechnol. 2017;13(4):192–208.
Polyhydroxyalkanoate (PHA) market by type (short chain length, medium chain length), production method (sugar fermentation, vegetable oil fermentation, methane fermentation), application, and region-global forecast to 2024. http://www.marketsandmarkets.com/Market-Reports/pha-market-395.html. Accessed 30 Mar 2020.
Shahzad K, Narodoslawsky M, Sagir M, Ali N, Ali S, Rashid MI, et al. Techno-economic feasibility of waste biorefinery: using slaughtering waste streams as starting material for biopolyester production. Waste Manag. 2017;67:73–85.
Papapostolou A, Karasavvas E, Chatzidoukas C. Oxygen mass transfer limitations set the performance boundaries of microbial PHA production processes–a model-based problem investigation supporting scale-up studies. Biochem Eng J. 2019;148:224–38.
Bandaiphet C, Prasertsan P. Effect of aeration and agitation rates and scale-up on oxygen transfer coefficient, kLa in exopolysaccharide production from Enterobacter cloacae WD7. Carbohydr Polym. 2006;66:216–28.
All authors wants to thank Defence Research & Development Organization Headquarter, New Delhi, India, for providing all the facilities.
Conflict of Interest
The authors declare that they have no conflict of interest.
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
This article is part of the Topical Collection on Bioconversion
About this article
Cite this article
Gahlawat, G., Kumari, P. & Bhagat, N.R. Technological Advances in the Production of Polyhydroxyalkanoate Biopolymers. Curr Sustainable Renewable Energy Rep (2020). https://doi.org/10.1007/s40518-020-00154-4
- Organic waste
- High throughput
- Metabolic engineering
- Downstream recovery