Limonene is a widely used monoterpene in the production of food, pharmaceuticals, biofuels, etc. The objective of this work was to engineer Rhodosporidium toruloides as a cell factory for the production of limonene.By overexpressing the limonene synthase (LS), neryl pyrophosphate synthase (NPPS)/geranyl pyrophosphate synthase and the native hydroxy-methyl-glutaryl-CoA reductase (HMGR), we established a baseline for limonene production based on the mevalonate route in Rhodosporidium toruloides. To further enhance the limonene titer, the acetoacetyl-CoA thiolase/HMGR (EfMvaE) and mevalonate synthase (EfMvaS) from Enterococcus faecalis, the mevalonate kinase from Methanosarcina mazei (MmMK) and the chimeric enzyme NPPS-LS were introduced in the carotenogenesis-deficient strain. The resulting strains produced a maximum limonene titer of 393.5 mg/L.In this study, we successfully engineered the carotenogenesis yeast R. toruloides to produce limonene. This is the first report on engineering R. toruloides toward limonene production based on NPP and the fusion protein SltNPPS-CltLS. The results demonstrated that R. toruloides is viable for limonene production, which would provide insights into microbial production of valuable monoterpenes.

Sasa Liu

Mengyao Zhang

Yuyao Ren

Guojie Jin

Yongsheng Tao

Liting Lyu

Zongbao K. Zhao

Xiaobing Yang

Biotechnology for Biofuels

BIOTECHNOLOGY FOR BIOFUELS 是一本开放获取的同行评议期刊，以描述生物燃料、化学品和其他生物产品生产中的技术和操作进步的高质量研究为特色。该杂志强调理解和推进生物技术和协同操作的应用，以改进植物和生物转化系统，以利用生物质、生物质衍生的中间体或二氧化碳进行生物生产这些产品，以及生物不可或缺的上游或下游操作生物质转化。生物燃料的生物技术专注于以下领域：• 陆地植物原料的开发• 藻类原料的开发• 用于生物转化的生物质预处理、分馏和提取• 酶工程、生产和分析• 细菌遗传学、生理学和代谢工程•真菌/酵母遗传学、生理学和代谢工程• 发酵、生物催化转化和反应动力学• 从生物质中生物生产化学品和生物产品• 厌氧消化、生物氢和生物电• 生物工艺集成、技术经济分析、建模和政策• L IFE循环评估和环境影响分析

Biotechnology for Biofuels is an open-access, peer-reviewed journal that features high-quality research describing technological and operational advancements in the production of biofuels, chemicals, and other bioproducts. The journal emphasizes understanding and advancing the applications of biotechnology and collaborative operations to improve plant and bioconversion systems for the biological production of these products using biomass, biomass-derived intermediates, or carbon dioxide, as well as the upstream and downstream operations essential for biomass conversion. The biotechnology of biofuels focuses on the following areas: • Development of terrestrial plant feedstocks • Development of algal feedstocks • Pretreatment, fractionation, and extraction of biomass for bioconversion • Enzyme engineering, production, and analysis • Bacterial genetics, physiology, and metabolic engineering • Fungal/yeast genetics, physiology, and metabolic engineering • Fermentation, biocatalytic conversion, and reaction kinetics • Bioproduction of chemicals and bioproducts from biomass • Anaerobic digestion, biohydrogen, and bioelectricity • Integration of bioprocesses, techno-economic analysis, modeling, and policy • Life cycle assessment and environmental impact analysis.

《Biotechnology for Biofuels》是一本开放获取的同行评审期刊，专注于生物燃料、化学品及其他生物产品生产中的技术与操作进展。该期刊强调理解和推动生物技术及协同操作的应用，以改进植物和生物转化系统，利用生物质、生物质衍生的中间体或二氧化碳进行生物生产。这些研究涵盖了生物质转化的上游和下游操作。生物燃料的生物技术主要集中在以下领域：• 陆地植物原料的开发 • 藻类原料的开发 • 生物质的预处理、分馏和提取 • 酶工程、生产与分析 • 细菌的遗传学、生理学和代谢工程 • 真菌/酵母的遗传学、生理学和代谢工程 • 发酵、生物催化转化和反应动力学 • 从生物质中生物生产化学品和生物产品 • 厌氧消化、生物氢和生物电 • 生物工艺集成、技术经济分析、建模和政策 • 生命周期评估和环境影响分析

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Engineering Rhodosporidium toruloides for limonene production

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