Yu Deng1, Mingxue Sun2, Sha Xu2, Jingwen Zhou2,3. 1. National Engineering Laboratory for Cereal Fermentation Technology (NELCF), Jiangnan University, Wuxi, Jiangsu, China. 2. Key Laboratory of Industrial Biotechnology, Ministry of Education and School of Biotechnology, Jiangnan University, Wuxi, Jiangsu, China. 3. Synergetic Innovation Center of Food Safety and Nutrition, Wuxi, Jiangsu, China.
Abstract
AIMS: In order to improve the availability of geranyl diphosphate (GPP) in the mevalonate pathway for enhancing (S)-linalool production in Saccharomyces cerevisiae. METHODS AND RESULTS: A (S)-linalool synthase (LIS): AaLS1 from Actinidia arguta was coexpressed with FPPS with different peptide linkers to redirect the flux from geranyl diphosphate (GPP) to (S)-linalool production in S. cerevisiae. The strain with the best peptide linker ((GGGGS)3 ), produced 101·55 ± 2·97 μg l(-1) (S)-linalool, a 69·7% increase compared to those with two independent LIS and FPPS expressed. In a 3-l fermenter, the (S)-linalool titre was further improved to 240·64 ± 5·31 μg l(-1) . CONCLUSIONS: The results demonstrate that the fusion proteins catalysing consecutive steps in a metabolic pathway significantly improved the (S)-linalool production with GPP as precursor. SIGNIFICANCE AND IMPACT OF THE STUDY: The fusion protein strategy co-expressing AaLS1 and FPPS, assembled with a long peptide linker made S. cerevisiae produced the highest reported (S)-Linalool titre to date.
AIMS: In order to improve the availability of geranyl diphosphate (GPP) in the mevalonate pathway for enhancing (S)-linalool production in Saccharomyces cerevisiae. METHODS AND RESULTS: A (S)-linalool synthase (LIS): AaLS1 from Actinidia arguta was coexpressed with FPPS with different peptide linkers to redirect the flux from geranyl diphosphate (GPP) to (S)-linalool production in S. cerevisiae. The strain with the best peptide linker ((GGGGS)3 ), produced 101·55 ± 2·97 μg l(-1) (S)-linalool, a 69·7% increase compared to those with two independent LIS and FPPS expressed. In a 3-l fermenter, the (S)-linalool titre was further improved to 240·64 ± 5·31 μg l(-1) . CONCLUSIONS: The results demonstrate that the fusion proteins catalysing consecutive steps in a metabolic pathway significantly improved the (S)-linalool production with GPP as precursor. SIGNIFICANCE AND IMPACT OF THE STUDY: The fusion protein strategy co-expressing AaLS1 and FPPS, assembled with a long peptide linker made S. cerevisiae produced the highest reported (S)-Linalool titre to date.
Authors: Behnaz Nowrouzi; Rachel A Li; Laura E Walls; Leo d'Espaux; Koray Malcı; Lungang Liang; Nestor Jonguitud-Borrego; Albert I Lerma-Escalera; Jose R Morones-Ramirez; Jay D Keasling; Leonardo Rios-Solis Journal: Microb Cell Fact Date: 2020-11-02 Impact factor: 5.328
Authors: Nicole G H Leferink; Adrian J Jervis; Ziga Zebec; Helen S Toogood; Sam Hay; Eriko Takano; Nigel S Scrutton Journal: ChemistrySelect Date: 2016-06-21 Impact factor: 2.109