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Enhancing the Production of Polyhydroxybutyrate, a Biodegradable Polymer by an Optimized Process Using a Novel Klebsiella Pneumonia Strain

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Abstract

Polyhydroxybutyrate (PHB), a sustainable biodegradable polymer with various industrial and medical applications, produced by microorganisms is a good replacement of conventional plastics, a major environmental contaminant. The aim of this study is to isolate and evaluate the PHB producing ability of the microorganisms isolated from various soil sources. By employing different staining methods such as Sudan black B, Nile red and Nile blue A, the primary screening of PHB producers was performed. Among the 104 soil isolates, four strains (SNA1, SNA2, SNA3 and SNA4) exhibited positive results for all the staining tests. On further studies, among these the strain SNA2 produced highest level of PHB (64% ) from its whole cell dry weight, while glucose (20 g/L) and urea (2 g/L) were used as the carbon and nitrogen sources respectively at pH 7, 30 °C after 48 h of incubation. The Fourier transform infrared spectroscopy (FTIR) and Proton Nuclear Magnetic Resonance (1H NMR) results confirmed that the polymer produced was PHB. The 16S rRNA sequencing revealed results that the strain SNA2 was Klebsiella pneumonia. The production ability of the novel Klebsiella pneumonia SNA2 was further enhanced by optimizing various process parameters such as carbon and nitrogen sources, pH, temperature and incubation period. The process optimization resulted in enhanced PHB synthesis (up to 75%) with Xylose as carbon source and urea as the nitrogen source with pH 7.5 at 35 °C after 36 h. In addition, an attempt to produce a biofilm from the extracted polymer was also successful. Thus, the novel bacterial isolate Klebsiella pneumonia SNA2 was proved to be a challenging host for the synthesis of PHB and the scale up studies are under progress. Whole genome sequencing, identification of pathway genes and metabolic engineering strategies were believed to enhance the PHB synthesis further to the industrial scale.

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Acknowledgements

The authors would like to thank Dr. U. Koushi Kumar for his help and constant support in the analysis part. Authors would also like to acknowledge all the faculty in department of Biotechnology, Vel Tech Rangarajan Dr. Sagunthala R&D Institute of Science and Technology, for the help extended for the research.

Funding

The part of this research was supported by the seed fund provided by office of Dean R&D, Vel Tech Rangarajan Dr. Sagunthala R&D Institute of Science and Technology.

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  1. Department of Biotechnology, Vel Tech Rangarajan Dr Sagunthala R&D Institute of Science and Technology, Chennai, Tamil Nadu, India

    Nirmala Nithya Raju & Mugesh Sankaranarayanan

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  1. Nirmala Nithya Raju
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Both the authors contributed to the study conception and design. Material preparation, data collection and analysis were performed by NNR. The first draft of the manuscript was written by NNR and the corrections and modifications were done by MS. Both the authors read and approved the final manuscript.

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Correspondence to Mugesh Sankaranarayanan.

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Raju, N.N., Sankaranarayanan, M. Enhancing the Production of Polyhydroxybutyrate, a Biodegradable Polymer by an Optimized Process Using a Novel Klebsiella Pneumonia Strain. Top Catal 67, 181–191 (2024). https://doi.org/10.1007/s11244-023-01877-6

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