Tuning Geraniol Biosynthesis via a Novel Decane-Responsive Promoter in Candida glycerinogenes.

Geraniol is a rose-scented monoterpene with significant commercial and industrial value in medicine, condiments, cosmetics, and bioenergy. Here, we first targeted geraniol as a reporter metabolite and explored the suitability and potential of Candida glycerinogenes as a heterologous host for monoterpenoid production. Subsequently, dual-pathway engineering was employed to improve the production of geraniol with a geraniol titer of 858.4 mg/L. We then applied a synthetic hybrid promoter approach to develop a decane-responsive hybrid promoter based on the native promoter PGAP derived from C. glycerinogenes itself. The hybrid promoter was able to be induced by n-decane with 3.6 times higher transcriptional intensity than the natural promoter PGAP. In particular, the hybrid promoter effectively reduces the conflict between cell growth and product formation in the production of geraniol. Ultimately, 1194.6 mg/L geraniol was obtained at the shake flask level. The strong and tunable decane-responsive hybrid promoter developed in this study provides an important tool for fine regulation of toxic terpenoid production in cells.

[1]  Nannan Xu,et al.  Development of a Highly Efficient Copper-Inducible GAL Regulation System (CuIGR) in Saccharomyces cerevisiae. , 2021, ACS synthetic biology.

[2]  Jifeng Yuan,et al.  High-Yielding Terpene-Based Biofuel Production in Rhodobacter capsulatus. , 2021, ACS synthetic biology.

[3]  Weiguo Zhang,et al.  Dual Regulation of Cytoplasm and Peroxisomes for Improved Α-Farnesene Production in Recombinant Pichia pastoris. , 2021, ACS synthetic biology.

[4]  M. Marchisio,et al.  Novel S. cerevisiae Hybrid Synthetic Promoters Based on Foreign Core Promoter Sequences , 2021, International journal of molecular sciences.

[5]  Xun Li,et al.  Engineering Escherichia coli for production of geraniol by systematic synthetic biology approaches and laboratory-evolved fusion tags. , 2021, Metabolic engineering.

[6]  J. Qiao,et al.  Characterization of trans-Nerolidol Synthase from Celastrus angulatus Maxim and Production of trans-Nerolidol in Engineered Saccharomyces cerevisiae. , 2021, Journal of agricultural and food chemistry.

[7]  Q. Hua,et al.  Biosynthesis of α-Pinene by Genetically Engineered Yarrowia lipolytica from Low-Cost Renewable Feedstocks. , 2020, Journal of agricultural and food chemistry.

[8]  S. Kampranis,et al.  Transforming yeast peroxisomes into microfactories for the efficient production of high-value isoprenoids , 2020, Proceedings of the National Academy of Sciences.

[9]  Xinyao Lu,et al.  Genetic engineering of an industrial yeast Candida glycerinogenes for efficient production of 2-phenylethanol , 2020, Applied Microbiology and Biotechnology.

[10]  G. Stephanopoulos,et al.  Enhancing isoprenoid synthesis in Yarrowia lipolytica by expressing the isopentenol utilization pathway and modulating intracellular hydrophobicity. , 2020, Metabolic engineering.

[11]  J. Keasling,et al.  Promoter Architecture and Promoter Engineering in Saccharomyces cerevisiae , 2020, Metabolites.

[12]  Shulin Chen,et al.  Expanding Toolbox for Genes Expression of Yarrowia lipolytica to Include Novel Inducible, Repressible and Hybrid Promoters. , 2020, ACS synthetic biology.

[13]  Xinyao Lu,et al.  Transcription factor Hap5 induces gsh2 expression to enhance 2-phenylethanol tolerance and production in an industrial yeast Candida glycerinogenes , 2020, Applied Microbiology and Biotechnology.

[14]  P. Ralph,et al.  Extrachromosomal genetic engineering of the marine diatom Phaeodactylum tricornutum enables the heterologous production of monoterpenoids. , 2020, ACS synthetic biology.

[15]  M. De Mey,et al.  Modulating transcription through development of semi-synthetic yeast core promoters , 2019, PloS one.

[16]  Xinyao Lu,et al.  Establishment of a transient CRISPR-Cas9 genome editing system in Candida glycerinogenes for co-production of ethanol and xylonic acid. , 2019, Journal of bioscience and bioengineering.

[17]  J. Qiao,et al.  Systematic Optimization of Limonene Production in Engineered Escherichia coli. , 2019, Journal of agricultural and food chemistry.

[18]  Jeffrey M. Skerker,et al.  Monoterpene production by the carotenogenic yeast Rhodosporidium toruloides , 2019, Microbial Cell Factories.

[19]  Gavin J. Williams,et al.  An Artificial Pathway for Isoprenoid Biosynthesis Decoupled from Native Hemiterpene Metabolism. , 2019, ACS synthetic biology.

[20]  G. Stephanopoulos,et al.  Two-step pathway for isoprenoid synthesis , 2018, Proceedings of the National Academy of Sciences.

[21]  Xie Jun,et al.  Pharmacological Properties of Geraniol – A Review , 2018, Planta Medica.

[22]  B. Møller,et al.  Phototrophic production of heterologous diterpenoids and a hydroxy-functionalized derivative from Chlamydomonas reinhardtii. , 2018, Metabolic engineering.

[23]  Xinyao Lu,et al.  Gene expression profiles of Candida glycerinogenes under combined heat and high-glucose stresses. , 2018, Journal of bioscience and bioengineering.

[24]  Jeffrey M. Skerker,et al.  Rhodosporidium toruloides: a new platform organism for conversion of lignocellulose into terpene biofuels and bioproducts , 2017, bioRxiv.

[25]  Ying Wang,et al.  Manipulation of GES and ERG20 for geraniol overproduction in Saccharomyces cerevisiae. , 2017, Metabolic engineering.

[26]  Yu Shen,et al.  Dynamic control of ERG20 expression combined with minimized endogenous downstream metabolism contributes to the improvement of geraniol production in Saccharomyces cerevisiae , 2017, Microbial Cell Factories.

[27]  H. Fang,et al.  Role of CgHOG1 in Stress Responses and Glycerol Overproduction of Candida glycerinogenes , 2016, Current Microbiology.

[28]  Qiang Hua,et al.  Heterologous production of α-farnesene in metabolically engineered strains of Yarrowia lipolytica. , 2016, Bioresource technology.

[29]  I. So,et al.  The antitumor effects of geraniol: Modulation of cancer hallmark pathways (Review) , 2016, International journal of oncology.

[30]  W. Whitman,et al.  Engineering the Autotroph Methanococcus maripaludis for Geraniol Production. , 2016, ACS synthetic biology.

[31]  Yu Shen,et al.  Improving monoterpene geraniol production through geranyl diphosphate synthesis regulation in Saccharomyces cerevisiae , 2016, Applied Microbiology and Biotechnology.

[32]  Ranran Zhang,et al.  The CYP51F1 Gene of Leptographium qinlingensis: Sequence Characteristic, Phylogeny and Transcript Levels , 2015, International journal of molecular sciences.

[33]  H. Woo,et al.  Microbial Synthesis of Myrcene by Metabolically Engineered Escherichia coli. , 2015, Journal of agricultural and food chemistry.

[34]  Seon-Won Kim,et al.  Geranyl diphosphate synthase: an important regulation point in balancing a recombinant monoterpene pathway in Escherichia coli. , 2015, Enzyme and microbial technology.

[35]  Seon-Won Kim,et al.  Engineering Escherichia coli for selective geraniol production with minimized endogenous dehydrogenation. , 2014, Journal of biotechnology.

[36]  M. Maffei,et al.  Engineering monoterpene production in yeast using a synthetic dominant negative geranyl diphosphate synthase. , 2014, ACS synthetic biology.

[37]  Jian Chen,et al.  Overproduction of geraniol by enhanced precursor supply in Saccharomyces cerevisiae. , 2013, Journal of biotechnology.

[38]  G. Stephanopoulos,et al.  Metabolic engineering: past and future. , 2013, Annual review of chemical and biomolecular engineering.

[39]  H. Fang,et al.  Cloning and characterization of a novel NAD+‐dependent glyceraldehyde‐3‐phosphate dehydrogenase gene from Candida glycerinogenes and use of its promoter , 2013, Yeast.

[40]  R. Fukuda,et al.  Transcriptional repression by glycerol of genes involved in the assimilation of n-alkanes and fatty acids in yeast Yarrowia lipolytica. , 2013, FEMS yeast research.

[41]  D. P. de Sousa,et al.  A Review on Anti-Inflammatory Activity of Monoterpenes , 2013, Molecules.

[42]  Rishi Garg,et al.  Controlling promoter strength and regulation in Saccharomyces cerevisiae using synthetic hybrid promoters , 2012, Biotechnology and bioengineering.

[43]  N. D. Da Silva,et al.  Introduction and expression of genes for metabolic engineering applications in Saccharomyces cerevisiae. , 2012, FEMS yeast research.

[44]  Eui-Sung Choi,et al.  Metabolic engineering of Escherichia coli for α-farnesene production. , 2011, Metabolic engineering.

[45]  J. Keasling,et al.  Engineering microbial biofuel tolerance and export using efflux pumps , 2011, Molecular systems biology.

[46]  Jens Nielsen,et al.  Characterization of different promoters for designing a new expression vector in Saccharomyces cerevisiae , 2010, Yeast.

[47]  Weiyang Chen,et al.  Geraniol — A review of a commercially important fragrance material , 2010 .

[48]  Jay D Keasling,et al.  Advanced biofuel production in microbes , 2010, Biotechnology journal.

[49]  Zhengxiang Wang,et al.  Cloning and characterization of a NAD+-dependent glycerol-3-phosphate dehydrogenase gene from Candida glycerinogenes, an industrial glycerol producer. , 2008, FEMS yeast research.

[50]  Timothy S. Ham,et al.  Production of the antimalarial drug precursor artemisinic acid in engineered yeast , 2006, Nature.

[51]  J. Keasling,et al.  Engineering a mevalonate pathway in Escherichia coli for production of terpenoids , 2003, Nature Biotechnology.

[52]  H. Fang,et al.  Glycerol production by a novel osmotolerant yeast Candida glycerinogenes , 2001, Applied Microbiology and Biotechnology.