Abstract
Poly-3-hydroxybutyrate (PHB) is a bio-based and biodegradable polymer produced by microbial fermentation from wide variety of feedstocks. Methanotrophic organism Methylosinus trichosporium has been reported for the production of PHB using alternate feedstock like methane which is economical and abundantly available. PHB was produced from methane by cultivating M. trichosporium in a bioreactor with continuous cell recycle system. Different gas sparging strategies were evaluated for impact on cell biomass and PHB production. The fermentation was conducted in different modes including batch and continuous culture with and without membrane recycle. Using membrane based recycle system the biomass increased from 1.5 to maximum of 7.31 g DCW/L in 27 days. Although PHB content was comparable in both batch mode and membrane based recycle system, a 4.9 fold increase in biomass production enhanced the PHB titer by 6.1 fold as compared to batch culture. Continuous cell recycle processes provide an opportunity to increase the competitiveness of gas fermentation based processes for methanotrophs based PHB production.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
Availability of Data and Materials
Data will be provided on request.
References
Cantera, S., R. Munoz, R. Lebrero, J. C. Lopez, Y. Rodriguez, and P. A Garcia-Encina (2018) Technologies for the bioconversion of methane into more valuable products. Curr. Opin. Biotechnol. 50: 128–135.
Garcia-Perez, T., J. C. Lopez, F. Passos, R. Lebrero, S. Revah, and R. Munoz (2018) Simultaneous methane abatement and PHB production by Methylocystis hirsuta in a novel gas-recycling bubble column bioreactor. Chem. Eng. J. 334: 691–697.
Hwang, I. Y., D. H. Hur, J. H. Lee, C. H. Park, I. S. Chang, J. W. Lee, and E. Y. Lee (2015) Batch conversion of methane to methanol using Methylosinus trichosporium OB3b as biocatalyst. J. Microbiol. Biotechnol. 25: 375–380.
Zhang, T., J. Zhou, X. Wang, and Y. Zhang (2017) Coupled effects of methane monooxygenase and nitrogen source on growth and poly-β-hydroxybutyrate (PHB) production of Methylosinus trichosporium OB3b. J. Environ. Sci. (China) 52: 49–57.
Asenjo, J. A. and J. S. Suk (1986) Microbial conversion of methane into poly-β-hydroxybutyrate (PHB): growth and intracellular product accumulation in a type II methanotroph. J. Ferment. Technol. 64: 271–278.
Myung, J., M. Kim, M. Pan, C. S. Criddle, and S. K. Y. Tang (2016) Low energy emulsion-based fermentation enabling accelerated methane mass transfer and growth of poly(3-hydroxybutyrate)-accumulating methanotrophs. Bioresour. Technol. 207: 302–307.
Wendlandt, K. D., M. Jechorek, J. Helm, and U. Stottmeister (1998) Production of PHB with a high molecular mass from methane. Polym. Degrad. Stab. 59: 191–194.
Kulkarni, P. P., S. B. Chavan, M. S. Deshpande, D. Sagotra, P. S. Kumbhar, and A. R. Ghosalkar (2022) Enrichment of Methylocystis dominant mixed culture from rice field for PHB production. J. Biotechnol. 343: 62–70.
Dos Santos Rodrigues, A. and B. V. E Andréa Medeiros Salgado (2009) Analysis of methane biodegradation by Methylosinus trichosporium OB3b. Braz. J. Microbiol. 40: 301–307.
Kulkarni, P. P., V. K. Khonde, M. S. Deshpande, T. R. Sabale, P. S. Kumbhar, and A. R. Ghosalkar (2021) Selection of methanotrophic platform for methanol production using methane and biogas. J. Biosci. Bioeng. 132: 460–468.
Shah, N. N., M. L. Hanna, and R. T. Taylor (1996) Batch cultivation of Methylosinus trichosporium OB3b: V. characterization of poly-β-hydroxybutyrate production under methane-dependent growth conditions. Biotechnol. Bioeng. 49: 161–171.
Henard, C. A., H. Smith, N. Dowe, M. G. Kalyuzhnaya, P. T. Pienkos, and M. T. Guarnieri (2016) Bioconversion of methane to lactate by an obligate methanotrophic bacterium. Sci. Rep. 6: 21585.
Sheets, J. P., X. Ge, Y. F. Li, Z. Yu, and Y. Li (2016) Biological conversion of biogas to methanol using methanotrophs isolated from solid-state anaerobic digestate. Bioresour. Technol. 201: 50–57.
Chidambarampadmavathy, K., O. P. Karthikeyan, and K. Heimann (2017) Sustainable bio-plastic production through landfill methane recycling. Renew. Sustain. Energy Rev. 71: 555–562.
Strong, P. J., B. Laycock, S. N. S. Mahamud, P. D. Jensen, P. A. Lant, G. Tyson, and S. Pratt (2016) The opportunity for highperformance biomaterials from methane. Microorganisms 4: 11.
Rahnama, F., E. Vasheghani-Farahani, F. Yazdian, and S. A. Shojaosadati (2012) PHB production by Methylocystis hirsuta from natural gas in a bubble column and a vertical loop bioreactor. Biochem. Eng. J. 65: 51–56.
Akaraonye, E., T. Keshavarz, and I. Roy (2010) Production of polyhydroxyalkanoates: The future green materials of choice. J. Chem. Technol. Biotechnol. 85: 732–743.
Ghoddosi, F., H. Golzar, F. Yazdian, K. Khosravi-Darani, and E. Vasheghani-Farahani (2019) Effect of carbon sources for PHB production in bubble column bioreactor: emphasis on improvement of methane uptake. J. Environ. Chem. Eng. 7: 102978.
Choi, J.-il and S. Y. Lee (1997) Process analysis and economic evaluation for poly(3-hydroxybutyrate) production by fermentation. Bioprocess. Eng. 17: 335–342.
Zaldivar Carrillo, J. A., L. Y. Stein, and D. Sauvageau (2018) Defining nutrient combinations for optimal growth and polyhydroxybutyrate production by Methylosinus trichosporium OB3b using response surface methodology. Front. Microbiol. 9: 1513.
Khosravi-Darani, K., F. Yazdian, F. Babapour, and A. R. Amirsadeghi (2019) Poly(3-hydroxybutyrate) production from natural gas by a methanotroph native bacterium in a bubble column bioreactor. Chem. Biochem. Eng. Q. 33: 69–77.
Salem, R., M. Soliman, A. Fergala, F. G. Audette, and A. ElDyasti (2021) Screening for methane utilizing mixed communities with high polyhydroxybutyrate (PHB) production capacity using different design approaches. Polymers 13: 1579.
Sabale, T., M. Babu, S. Panchwagh, and A. Ghosalkar (2021) Continuous culture of Nitrosomonas eutropha and biomethanol production using whole cell biotransformation. J. Chem. Technol. Biotechnol. 96: 1411–1418.
Wendlandt, K. D., M. Jechorek, J. Helm, and U. Stottmeister (2001) Producing poly-3-hydroxybutyrate with a high molecular mass from methane. J. Biotechnol. 86: 127–133.
Zhang, Y., J. Xin, L. Chen, and C. Xia (2009) The methane monooxygenase intrinsic activity of kinds of methanotrophs. Appl. Biochem. Biotechnol. 157: 431–441.
Strong, P. J., S. Xie, and W. P. Clarke (2015) Methane as a resource: can the methanotrophs add value? Environ. Sci. Technol. 49: 4001–4018.
Cui, P., Y. Shao, Y. Wang, R. Zhao, H. Zhan, and W. Zhong (2021) Co-production of polyhydroxybutyrate (PHB) and coenzyme Q10 (CoQ10) via no-sugar fermentation-a case by Methylobacterium sp. XJLW. Ann. Microbiol. 71: 20.
Patel, S. K. S., P. Mardina, D. Kim, S. Y. Kim, V. C. Kalia, I. W. Kim, and J. K. Lee (2016) Improvement in methanol production by regulating the composition of synthetic gas mixture and raw biogas. Bioresour. Technol. 218: 202–208.
Lee, E. H., K. E. Moon, T. G. Kim, S. D. Lee, and K. S. Cho (2015) Inhibitory effects of sulfur compounds on methane oxidation by a methane-oxidizing consortium. J. Biosci. Bioeng. 120: 670–676.
Soni, B. K., J. Conrad, R. L. Kelley, and V. J. Srivastava (1998) Effect of temperature and pressure on growth and methane utilization by several methanotrophic cultures. Appl. Biochem. Biotechnol. 70: 729–738.
Han, B., T. Su, H. Wu, Z. Gou, X. H. Xing, H. Jiang, Y. Chen, X. Li, and J. C. Murell (2009) Paraffin oil as a “methane vector” for rapid and high cell density cultivation of Methylosinus trichosporium OB3b. Appl. Microbiol. Biotechnol. 83: 669–677.
Pen, N., L. Soussan, M. P. Belleville, J. Sanchez, and D. Paolucci-Jeanjean (2016) Methane hydroxylation by Methylosinus trichosporium OB3b: monitoring the biocatalyst activity for methanol production optimization in an innovative membrane bioreactor. Biotechnol. Bioprocess Eng. 21: 283–293.
Taylor, A., P. Molzahn, T. Bushnell, C. Cheney, M. LaJeunesse, M. Azizian, and L. Semprini (2018) Immobilization of Methylosinus trichosporium OB3b for methanol production. J. Ind. Microbiol. Biotechnol. 45: 201–211.
Zhang, T., J. Zhou, X. Wang, and Y. Zhang (2019) Poly-β-hydroxybutyrate production by Methylosinus trichosporium OB3b at different gas-phase conditions. Iran J. Biotechnol. 17: e1866.
Lee, J., N. Jang, M. Yasin, E. Y. Lee, I. S. Chang, and C. Kim (2016) Enhanced mass transfer rate of methane via hollow fiber membrane modules for Methylosinus trichosporium OB3b fermentation. J. Ind. Eng. Chem. 39: 149–152.
Acknowledgments
The authors are thankful to Mr. Sachin Mane and Mr. Devesh Mahabare from Analytical Science Center of Praj Matrix - R&D Center for providing gas chromatography support.
Funding
The authors gratefully acknowledge Praj Industries Ltd. Pune for financial support to conduct the research work.
Author information
Authors and Affiliations
Contributions
TRS: Conceived, design of research and experiments, performed experiment, samples analysis for PHB, data analysis, manuscript writing. PPK: Samples analysis for PHB, design of research, performed enzyme assay, data analysis, manuscript writing. ARG: Supervised and designed research, data analysis corrected and reviewed manuscript. All authors read and approved the manuscript.
Corresponding author
Ethics declarations
This article does not contain any studies with human participants or animals performed by any of the authors. The authors declare that they have no competing interests.
Additional information
Publisher’s Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Consent to Participate
Not applicable
Consent for Publication
Not applicable
Rights and permissions
About this article
Cite this article
Sabale, T.R., Kulkarni, P.P. & Ghosalkar, A.R. Methane Based Continuous Culture of Methylosinus trichosporium for Production of Poly-3-hydroxybutyrate Using Membrane Recycle System. Biotechnol Bioproc E 28, 519–527 (2023). https://doi.org/10.1007/s12257-022-0383-3
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s12257-022-0383-3