This project will use protein engineering to improve enzymes required for the efficient production of isoprene from renewable resources. Isoprene is an industrial chemical widely used in the polymer industry, and mainly produced from petrochemical feedstocks.
Sustainable solutions for isoprene synthesis have been proposed, which rely on the engineering of biosynthetic isoprene pathways in microbes. These pathways use isoprene synthase (IspS) to convert dimethylallyl pyrophosphate (DMAPP) into isoprene. IspS is relatively inefficient, with low turnover and high KM, as well as poor soluble microbial expression. To achieve commercially relevant yields of isoprene, improvement of IspS is necessary.
The aim of this project is to develop an isoprene synthase with improved solubility and catalytic efficiency, to be integrated into an engineered pathway for the microbial production of isoprene. First, you will investigate the isoprene production ability of a range of plant isoprene synthase candidates. Codon optimised IspS genes will be synthesised and their soluble expression in E. coli will be investigated. After identification of a suitable candidate, site-directed mutagenesis based on homology models will be used to improve catalytic activity and solubility. Medium-sized libraries will be screened on lysates for the conversion of DMAPP to isoprene, using established GC methods.
The project will involve skills relevant to enzyme engineering for biotechnology applications, including molecular biology and microbiology, protein expression and purification, computational methods, and competence in analytical techniques (especially HPLC, GC, GCMS). Training will be provided where necessary. Good communication skills are important.
This PhD project is part of a University-funded Doctoral Training Programme in Synthetic Biology and associated with Nottingham’s new BBSRC/EPSRC Synthetic Biology Research Centre. Students will benefit from a diverse range of training opportunities, including specialist workshops, lectures and seminars, as well as participation in Nottingham’s yearly BBSRC DTP Spring School event.
Applications are invited from UK/EU nationals with a background in chemistry, biochemistry, (bio)chemical engineering or a related discipline, who have or expect to graduate with a first-class or good 2:1 honours degree, or a distinction or high merit at MSc level (or international equivalent). The studentship will cover full PhD tuition fees and a tax free stipend for four years for UK/EU students.
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Last updated July 31, 2017