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Hydroxytyrosol (HT) is a valuable aromatic compound with numerous applications. Herein, we enabled the efficient and scalable HT production in engineered () from glucose. Starting from a tyrosol-overproducing strain, six HpaB/HpaC combinations were investigated, and the best catalytic performance was acquired with HpaB from (HpaB) and HpaC from (HpaC), resulting in 425.7 mg/L HT in shake flasks. Next, weakening the tryptophan biosynthetic pathway through downregulating the expression of (encoding anthranilate synthase) further improved the HT titer by 27.2% compared to the base strain. Moreover, the cytosolic NADH supply was improved through introducing the feedback-resistant mutant of the TyrA (the NAD-dependent chorismate mutase/prephenate dehydrogenase, TyrA*) from , which further increased the HT titer by 36.9% compared to the base strain. The best performing strain was obtained by optimizing the biosynthesis of HT in through a screening for an effective HpaB/HpaC combination, biosynthetic flux rewiring, and cofactor engineering, which enabled the titer of HT reaching 1120.0 mg/L in the shake flask. Finally, the engineered strain produced 6.97 g/L of HT by fed-batch fermentation, which represents the highest titer for HT biosynthesis in microorganisms reported to date.
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http://dx.doi.org/10.1021/acssynbio.2c00316 | DOI Listing |
Front Bioeng Biotechnol
February 2025
Department of Molecular Science and Technology and Advanced College of Bio-convergence Engineering, Ajou University, Suwon, Republic of Korea.
The efficient identification of microbial strains capable of producing rare sphingoid bases, such as sphingosine and sphinganine, is critical for advancing microbial fermentation processes and addressing increasing industrial demands. , a non-conventional yeast, naturally overproduces tetraacetyl phytosphingosine (TAPS); however, the production of other valuable sphingoid bases, including sphingosine, sphinganine, and triacetyl sphingosine, remains a key target. In this study, we developed a novel screening method utilizing fluorescein sodium, a selective fluorescent dye that specifically reacts with non-acetylated sphingoid bases-sphinganine, sphingosine, and phytosphingosine-while exhibiting no reactivity with TAPS.
View Article and Find Full Text PDFSynth Syst Biotechnol
June 2025
Key Laboratory of Combinatorial Biosynthesis and Drug Discovery, Ministry of Education and School of Pharmaceutical Sciences, Wuhan University, 430072, Wuhan, Hubei Province, China.
Endusamycin, a naturally occurring polyether ionophore antibiotic, exhibits extensive antitumor activities. Despite its promising potential, the titer of endusamycin is significantly lower compared to widely used polyether compounds, and no reports have been published regarding its overproduction. In this study, various metabolic engineering strategies were performed to enhance endusamycin production.
View Article and Find Full Text PDFMetab Eng
March 2025
Microbiology Division, IBR (Instituto de Biología Molecular y Celular de Rosario), Consejo Nacional de Investigaciones Científicas y Técnicas, Facultad de Ciencias Bioquímicas y Farmacéuticas, Universidad Nacional de Rosario, Ocampo y Esmeralda, 2000 Rosario, Argentina. Electronic address:
Microbial fatty acids (FAs) hold significant potential as alternatives for the oleochemical industry. However, expanding the functional and structural diversity of microbial FA-derived products is essential to fully leverage this potential. Methyl-branched-chain FAs (MBFAs) are of particular interest as high-performance industrial compounds.
View Article and Find Full Text PDFJ Agric Food Chem
March 2025
State Key Laboratory of Food Science and Technology, School of Food Science and Technology, Jiangnan University, Wuxi 214122, China.
Chlorogenic acid (CGA) is a natural hydroxycinnamic acid ester with significant applications in food preservation and nutritional health. However, extraction of CGA from plants is challenging, resulting in low purity that fails to meet increasing market demands. Furthermore, the broad substrate specificity of hydroxycinnamoyl-CoA:quinic acid transferase catalysis generating a plethora of byproducts, lack of NADPH regeneration, and the presence of degrading proteins impede microbial synthesis of CGA.
View Article and Find Full Text PDFACS Synth Biol
February 2025
CAS Key Laboratory of Microbial Physiological and Metabolic Engineering, State Key Laboratory of Mycology, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, PR China.
δ-Valerolactam (VL), as an organic compound, is an important precursor chemical for nylon and has a wide range of applications in organic synthesis, pharmaceutical synthesis, polymer materials, and other fields. This study introduces a novel biosynthetic method for producing VL in the engineered strain BL21 through the reprogramming of polyketide synthases (PKS). Initially, an in vitro multienzyme system was constructed to verify the reliability of the VL synthesis pathway.
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