The heterologous production of terpenes by the thermophile Parageobacillus thermoglucosidasius in a consolidated bioprocess using waste bread |
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| Affiliation: | 1. Laboratory of Microbial Ecology and Technology (LETMI), National Institute of Applied Sciences and Technology (INSAT), University of Carthage, Tunisia;2. High School of Food Industry of Tunis (ESIAT), University of Carthage, Tunisia;3. Laboratoire de Traitement et de Valorisation des Rejets Hydriques (LTVRH), Center of Water Researches and Technologies (CERTE), University of Carthage, Tourist Route Soliman, BP 273-8020 Nabeul, Tunisia;1. Center for Process Systems Engineering, Department of Chemical Engineering, Imperial College London, South Kensington Campus, London SW7 2AZ, England, UK;2. Department of Biology & Biochemistry, University of Bath, Bath, Somerset, England, UK;1. DBT-ICGEB Center for Advanced Bioenergy Research, International Centre for Genetic Engineering and Biotechnology, Aruna Asaf Ali Marg, 110067 New Delhi, India;2. Department of Biological and Medical Sciences, Oxford Brookes University, Gipsy Lane, OX3 0BP, Oxford, UK;1. Institute of Molecular Enzyme Technology, Heinrich Heine University Düsseldorf, Forschungszentrum Jülich, Jülich, Germany;2. Institute of Bio- and Geosciences IBG-1, Forschungszentrum Jülich, Jülich, Germany;3. Central Division of Analytical Chemistry ZEA-3: Analytik/Biospec, Forschungszentrum Jülich, Jülich, Germany;4. Cluster of Excellence on Plant Sciences (CEPLAS), Germany;5. Bioeconomy Science Center (BioSC), Forschungszentrum Jülich, Jülich, Germany |
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| Abstract: | Parageobacillus thermoglucosidasius is a genetically tractable thermophile that grows rapidly at elevated temperatures, with a doubling time at 65 °C comparable to the shortest doubling times of Escherichia coli. It is capable of using a wide variety of substrates, including carbohydrate oligomers, and has been developed for the industrial production of ethanol. In this study, P. thermoglucosidasius NCIMB11955 has been engineered to produce the sesquiterpene τ-muurolol by introduction of a heterologous mevalonate pathway constructed using genes from several thermophilic archaea together with a recently characterised thermostable τ-muurolol synthase. P. thermoglucosidasius naturally uses the methylerythritol phosphate pathway for production of the terpene precursor, isopentenyl pyrophosphate, while archaea use a version of the mevalonate pathway. By introducing the orthogonal archaeal pathway it was possible to increase the flux through to sesquiterpene biosynthesis. Construction of such a large metabolic pathway created problems with genetic vector introduction and stability, so recombinant plasmids were introduced by conjugation, and a thermostable serine integrase system was developed for integration of large pathways onto the chromosome. Finally, by making the heterologous pathway maltose-inducible we demonstrate that the new strain is capable of using waste bread directly as an autoinduction carbon source for the production of terpenes in a consolidated bioprocess. |
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| Keywords: | Consolidated bioprocess Thermophile Terpenes |
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