Y-L Liang1, S-J Zhao1, L-X Xu1, X-Y Zhang1. 1. Department of Bioengineering, College of Biological and Agricultural Engineering, Jilin University, ChangChun, China.
Abstract
AIMS: Dammarenediol production by an engineered yeast Saccharomyces cerevisiae was investigated. METHODS AND RESULTS: A dammarenediol-producing engineered yeast was constructed by heterologous expression of the dammarenediol synthase gene from Panax ginseng hairy roots through RT-PCR. Fermentation was carried out in a 5-L GRJY-bioreactor with an inoculum size of 1% v/v at 30°C. Dammarenediol detection was performed with silica gel chromatography and HPLC. Determination of dammarenediol synthase activity subcellular distribution was carried out by surveying the enzyme activity in microsomes, lipid particles and total yeast homogenate. When cultured under aerobic conditions, the engineered yeast could produce dammarenediol up to 250μgl(-1). However, when an anaerobic shift strategy was employed, dammarenediol accumulated at a level as twice as that under aerobic condition. The dammarenediol synthase and dammarenediol were mainly localized in lipid particles. CONCLUSIONS: Dammarenediol could be heterologously produced in engineered yeast. The heterologously expressed dammarenediol synthase is mainly localized in lipid particles. Anaerobic shift strategy could enhance the dammarenediol level in the engineered yeast. SIGNIFICANCE AND IMPACT OF THE STUDY: This study showed that the high-value plant product dammarenediol could be produced by heterologous expression of the according gene in yeast. Furthermore, the anaerobic shift strategy could be potentially applied in oxidosqualene-derived compounds production in yeast. Here, the information about subcellular distribution of heterologously expressed dammarenediol synthase in the engineered yeast was also provided.
AIMS: Dammarenediol production by an engineered yeastSaccharomyces cerevisiae was investigated. METHODS AND RESULTS: A dammarenediol-producing engineered yeast was constructed by heterologous expression of the dammarenediol synthase gene from Panax ginseng hairy roots through RT-PCR. Fermentation was carried out in a 5-L GRJY-bioreactor with an inoculum size of 1% v/v at 30°C. Dammarenediol detection was performed with silica gel chromatography and HPLC. Determination of dammarenediol synthase activity subcellular distribution was carried out by surveying the enzyme activity in microsomes, lipid particles and total yeast homogenate. When cultured under aerobic conditions, the engineered yeast could produce dammarenediol up to 250μgl(-1). However, when an anaerobic shift strategy was employed, dammarenediol accumulated at a level as twice as that under aerobic condition. The dammarenediol synthase and dammarenediol were mainly localized in lipid particles. CONCLUSIONS: Dammarenediol could be heterologously produced in engineered yeast. The heterologously expressed dammarenediol synthase is mainly localized in lipid particles. Anaerobic shift strategy could enhance the dammarenediol level in the engineered yeast. SIGNIFICANCE AND IMPACT OF THE STUDY: This study showed that the high-value plant product dammarenediol could be produced by heterologous expression of the according gene in yeast. Furthermore, the anaerobic shift strategy could be potentially applied in oxidosqualene-derived compounds production in yeast. Here, the information about subcellular distribution of heterologously expressed dammarenediol synthase in the engineered yeast was also provided.
Authors: Jonathan Asmund Arnesen; Arian Belmonte Del Ama; Sidharth Jayachandran; Jonathan Dahlin; Daniela Rago; Aaron John Christian Andersen; Irina Borodina Journal: Metab Eng Commun Date: 2022-03-26
Authors: Anita Loeschcke; Dennis Dienst; Vera Wewer; Jennifer Hage-Hülsmann; Maximilian Dietsch; Sarah Kranz-Finger; Vanessa Hüren; Sabine Metzger; Vlada B Urlacher; Tamara Gigolashvili; Stanislav Kopriva; Ilka M Axmann; Thomas Drepper; Karl-Erich Jaeger Journal: PLoS One Date: 2017-12-27 Impact factor: 3.240