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Burkholderia xenovorans LB400 possesses a functional polyhydroxyalkanoate anabolic pathway encoded by the pha genes and synthesizes poly(3-hydroxybutyrate) under nitrogen-limiting conditions

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Abstract

Polyhydroxyalkanoates (PHAs) are biodegradable bioplastics that are synthesized by diverse bacteria. In this study, the synthesis of PHAs by the model aromatic-degrading strain Burkholderia xenovorans LB400 was analyzed. Twelve pha genes including three copies of phaC and five copies of the phasin-coding phaP genes are distributed among the three LB400 replicons. The phaC1ABR gene cluster that encodes the enzymes of the PHA anabolic pathway is located at chromosome 1 of strain LB400. During the growth of strain LB400 on glucose under nitrogen limitation, the expression of the phaC1, phaA, phaP1, phaR, and phaZ genes was induced. Under nitrogen limitation, PHA accumulation in LB400 cells was observed by fluorescence microscopy after Nile Red staining. GC-MS analyses revealed that the PHA accumulated under nitrogen limitation was poly(3-hydroxybutyrate) (PHB). LB400 cells grown on glucose as the sole carbon source under nitrogen limitation accumulated 40 ± 0.96% PHB of the cell dry weight, whereas no PHA was observed in cells grown in control medium. The functionality of the phaC1 gene from strain LB400 was further studied using heterologous expression in a Pseudomonas putida KT40C1ZC2 mutant strain derived from P. putida KT2440 that is unable to synthesize PHAs. Interestingly, KT40C1ZC2[pVNC1] cells that express the phaC1 gene from strain LB400 were able to synthesize PHB (33.5% dry weight). This study indicates that B. xenovorans LB400 possesses a functional PHA synthetic pathway that is encoded by the pha genes and is capable of synthesizing PHB.

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Acknowledgments

The authors thank Maria Auxiliadora Prieto for the generous support in heterologous expression assays and Vincent Collins for critical reading of the manuscript.

Funding

The present work was supported by PhD scholarship CONICYT 21100268 (VU), Programa de Investigación Asociativa (PIA) Anillo ACT172128 GAMBIO (MS), FONDECYT 1151174 and 1110992 (MS), USM 131342 and 131562 (MS), USM Pie>A (PV), USM CN&SB (MS), and CYTED-PRIBOP grants (MS).

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  1. Viviana Urtuvia and Pamela Villegas contributed equally to this work.

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  1. Laboratorio de Microbiología Molecular y Biotecnología Ambiental, Departamento de Química & Centro de Biotecnología, Universidad Técnica Federico Santa María, Avenida España 1680, 2390123, Valparaíso, Chile

    Viviana Urtuvia, Pamela Villegas, Sebastián Fuentes, Myriam González & Michael Seeger

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  1. Viviana Urtuvia
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  2. Pamela Villegas
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  3. Sebastián Fuentes
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Correspondence to Michael Seeger.

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Urtuvia, V., Villegas, P., Fuentes, S. et al. Burkholderia xenovorans LB400 possesses a functional polyhydroxyalkanoate anabolic pathway encoded by the pha genes and synthesizes poly(3-hydroxybutyrate) under nitrogen-limiting conditions. Int Microbiol 21, 47–57 (2018). https://doi.org/10.1007/s10123-018-0004-3

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