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Optimization of growth conditions for the biosynthesis of medium-chain length polyhydroxyalkanoates from Bacillus megaterium DSM 509: experimental analysis, statistical modelling, and characterization

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Abstract

Bioplastics are biobased materials produced from renewable sources like vegetable fats and oils, corn starch, straw, woodchips, sawdust, and recycled food wastes. Microbial polyhydroxyalkanoate (PHA), a type of bioplastics, has unique biocompatibility, non-toxicity, and biodegradability properties, making it suitable as an eco-friendly plastic material. Bacillus megaterium DSM 509 is known to produce short-chain length (scl) PHA. The present study explores the capability of Bacillus megaterium DSM 509 to produce a novel type of PHA, medium-chain length (mcl), and its mixture with scl-PHA. Statistical modelling and experimental results were used to optimize the growth conditions for enhanced production of cost-effective and biocompatible polyhydroxyoctanoates and polyhydroxydecanoates. A 6-run mixture design for glycerol and ammonium sulphate concentrations ratio was performed. A linear regression model was built based on experimental data, and the optimum conditions for different glycerol and nitrogen concentrations were derived and validated. A positive correlation (r2, 0.95, and 0.87) was observed, resulting in the model’s fitness. Optical density studies showed the impact of dissolved oxygen (DO) content on PHA production. It was observed that PHA production was enhanced for each sample at the optimum value of DO. The highest intracellular PHA was produced by microbial culture, having a C/N ratio of 25 (high glycerol and low nitrogen source) as detected on FTIR: peak height at 1740 cm−1 was 2.157. Results on gas chromatographic (GC–MS and GC-FID) analysis confirmed the presence of a mixture of scl and mcl-PHA as well as their monomer composition. The maximum PHA production obtained was 4.73 g.L−1 (60%) of dry cell weight at C/N ratio 25. The nature and physical form of bacterial cells was revealed using transmission electron microscopy (TEM). It is concluded that Bacillus megaterium is capable of producing a combination of scl-PHA and mcl-PHA, while feeding on nitrogen-deficient mineral medium.

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Acknowledgements

Facilities of SEM and FTIR are provided by Unité de Recherche ABTE (Aliments Bioprocédés Toxicologie Environnements), EA 4651, Esplanade de la Paix.

Funding

Higher Education Commission (HEC), Islamabad, Pakistan, supported this work.

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Authors and Affiliations

  1. Department of Biochemistry, Government College Women University, Faisalabad, 38040, Pakistan

    Salma Shahid, Sadia Noreen & Sofia Parveen

  2. Unité de Recherche ABTE (Aliments Bioprocédés Toxicologie Environnements), EA 4651, Esplanade de la Paix, Université de Caen Normandie, 14032 Cedex 5, Caen, France

    David Corroler, Ridha Mosrati, Caroline Amiel & Jean-Luc Guillard

  3. School of Life Science and Food Engineering, Huaiyin Institute of Technology, Huaian, 223003, China

    Muhammad Bilal

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  1. Salma Shahid
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Correspondence to Muhammad Bilal.

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Shahid, S., Corroler, D., Mosrati, R. et al. Optimization of growth conditions for the biosynthesis of medium-chain length polyhydroxyalkanoates from Bacillus megaterium DSM 509: experimental analysis, statistical modelling, and characterization. Biomass Conv. Bioref. 13, 12249–12264 (2023). https://doi.org/10.1007/s13399-021-01986-w

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  • DOI: https://doi.org/10.1007/s13399-021-01986-w

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