Metabolic engineering of Saccharomyces cerevisiae for efficient production of endocrocin and emodin

Sun, Lei; Liu, Guiyou; Li, Ya; Jiang, Dayong; Guo, Wenfeng; Xu, Hui; Zhan, Renya

doi:10.1016/j.ymben.2019.04.008

Cited by 29 publications

(28 citation statements)

References 47 publications

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“…1b). In addition, to facilitate the post-translational activation of the PKS, we chose two PPTases to activate Bik1: Ppt1 (accession number CCE73639), the native PPTase from F. fujikuroi, and NpgA (accession number AAF12814), a PPTase from A. nidulans, which has previously been successfully used in yeast to activate PKSs and nonribosomal peptide synthetases (NPRSs) 6,12,13 . Both PPTase genes were added into the constructed yeast bikaverin expression cassette under the control of P ENO2 and P TEF2 promoters, respectively.…”

Section: Resultsmentioning

confidence: 99%

“…Saccharomyces cerevisiae is one of the most widely used hosts for the metabolic engineering of natural products including antibiotics, terpenoids, cannabinoids, and opiates [6][7][8][9] . Despite the genetic tractability and scalability of metabolic engineering in yeast and a few examples of the successful biosynthesis of fungal polyketides in S. cerevisiae, the biosynthesis and large-scale fermentation of complex polyketides in yeast remains a challenge, due to the large size of the PKS biosynthetic enzymes, complicated biosynthetic pathways, and highly regulated nature of the process [10][11][12][13][14] .…”

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confidence: 99%

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Pathway engineering in yeast for synthesizing the complex polyketide bikaverin

Zhao

Yao

et al. 2020

Nat Commun

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Fungal polyketides display remarkable structural diversity and bioactivity, and therefore the biosynthesis and engineering of this large class of molecules is therapeutically significant. Here, we successfully recode, construct and characterize the biosynthetic pathway of bikaverin, a tetracyclic polyketide with antibiotic, antifungal and anticancer properties, in S. cerevisiae. We use a green fluorescent protein (GFP) mapping strategy to identify the low expression of Bik1 (polyketide synthase) as a major bottleneck step in the pathway, and a promoter exchange strategy is used to increase expression of Bik1 and bikaverin titer. Then, we use an enzyme-fusion strategy to directly couple the monooxygenase (Bik2) and methyltransferase (Bik3) to efficiently channel intermediates between modifying enzymes, leading to an improved titer of bikaverin at 202.75 mg/L with flask fermentation (273-fold higher than the initial titer). This study demonstrates that the biosynthesis of complex fungal polyketides can be established and efficiently engineered in S. cerevisiae, highlighting the potential for natural product synthesis and large-scale fermentation in yeast.

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Section: Resultsmentioning

confidence: 99%

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confidence: 99%

Pathway engineering in yeast for synthesizing the complex polyketide bikaverin

Zhao

Yao

et al. 2020

Nat Commun

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“…Sun et al reconstituted a biosynthetic pathway of emodin in Saccharomyces cerevisiae and confirmed that the biosynthetic pathway is polyketide pathway [ 15 ]. To provide an insight into the probable mechanism underlying the effect of KH 2 PO 4 adding on emodin production, we detected expression levels of 12 genes emoA, emoB, emoC, emoD, emoE, emoF, emoG, emoH, emoI, emoJ, emoK, and emoL of polyketide pathway by qRT-PCR.…”

Section: Resultsmentioning

confidence: 99%

Enhancement of Emodin Production by Medium Optimization and KH2PO4 Supplementation in Submerged Fermentation of Marine-Derived Aspergillus favipes HN4-13

et al. 2021

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Emodin is a widely distributed anthraquinone derivative with a variety of biological activities, one that can be efficiently produced by marine-derived fungus Aspergillus favipes HN4-13. However, its relatively low fermentation yield limits further development and pharmaceutical research work. In this study, Plaekett–Burman design and central composite design were adopted to optimize the fermentation conditions of A. favipes HN4-13. Optimal fermentation conditions in a 250-mL Erlenmeyer flask with 50 mL of medium were 59.3 g/L soluble starch, 10 g/L yeast extract paste, 30 g/L seawater salt, 1.04 g/L KH2PO4, 0.05 g/L MgSO4·7H2O, 0.01 g/L FeSO4·7H2O, seed culture 24 h, pH 5, inoculum size 18%, culture temperature 32 °C, and shaking at 160 rpm/min for 7 days. The production of emodin could achieve 132.40 ± 3.09 mg/L, with no significant difference from the predicted value (132.47 mg/L). Furthermore, KH2PO4 supplementation strategy was employed to regulate the mycelial morphology, upregulate the transcriptional level of biosynthesis gene cluster, and enhance emodin production (185.56 ± 4.39 mg/L).

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“…Besides, the de novo synthesis pathway of m-cresol was constructed from S. cerevisiae glucose by introducing the heterologous pathway of 6-MSA ( Hitschler and Boles, 2019 ). The high yield of emodin reached 528.4 mg/L in S. cerevisiae BJ5464-NPGA by heterologous reconstruction of the biosynthesis pathway of endorphin and emodin ( Sun et al, 2019 ).…”

Section: Introductionmentioning

confidence: 99%

Characterization and Heterologous Expression of UDP-Glucose 4-Epimerase From a Hericium erinaceus Mutant with High Polysaccharide Production

Zou

Ren

et al. 2021

Front. Bioeng. Biotechnol.

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Hericium erinaceus is an important medicinal fungus in traditional Chinese medicine because of its polysaccharides and other natural products. Compared terpenoids and polyketides, the analysis of synthetic pathway of polysaccharides is more difficult because of the many genes involved in central metabolism. In previous studies, A6180, encoding a putative UDP-glucose 4-epimerase (UGE) in an H. erinaceus mutant with high production of active polysaccharides, was significantly upregulated. Since there is no reliable genetic manipulation technology for H. erinaceus, we employed Escherichia coli and Saccharomyces cerevisiae to study the function and activity of A6180. The recombinant overexpression vector pET22b-A6180 was constructed for heterologous expression in E. coli. The enzymatic properties of the recombinant protein were investigated. It showed that the recombinant A6180 could strongly convert UDP-α-D-glucose into UDP-α-D-galactose under optimal conditions (pH 6.0, 30°C). In addition, when A6180 was introduced into S. cerevisiae BY4742, xylose was detected in the polysaccharide composition of the yeast transformant. This suggested that the protein coded by A6180 might be a multifunctional enzyme. The generated polysaccharides with a new composition of sugars showed enhanced macrophage activity in vitro. These results indicate that A6180 plays an important role in the structure and activity of polysaccharides. It is a promising strategy for producing polysaccharides with higher activity by introducing A6180 into polysaccharide-producing mushrooms.

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Metabolic engineering of Saccharomyces cerevisiae for efficient production of endocrocin and emodin

Cited by 29 publications

References 47 publications

Pathway engineering in yeast for synthesizing the complex polyketide bikaverin

Pathway engineering in yeast for synthesizing the complex polyketide bikaverin

Enhancement of Emodin Production by Medium Optimization and KH2PO4 Supplementation in Submerged Fermentation of Marine-Derived Aspergillus favipes HN4-13

Characterization and Heterologous Expression of UDP-Glucose 4-Epimerase From a Hericium erinaceus Mutant with High Polysaccharide Production

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