BIOORGANIC CHEMISTRY
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2024
·
IF 4.5
Shuyan Tang,Weiting Ji,Yunqiu Zhao,Jian Zhang,Dongzhi Wei,Feng-Qing Wang
Betulinic acid (BA) is a lupinane-type pentacyclic triterpenoid natural product derived from lupeol that has favorable anti-inflammatory and anti-tumor activities. Currently, BA is mainly produced via botanical extraction, which significantly limits its widespread use. In this study, we investigated the de novo synthesis of BA in Saccharomyces cerevisiae , and to facilitate the synthesis and storage of hydrophobic BA, we adopted a dual-engineering strategy involving peroxisomes and lipid droplets to construct the BA biosynthetic pathway. By expressing Betula platyphylla -derived lupeol C-28 oxidase (BPLO) and Arabidopsis -derived ATR1, we succeeded in developing a BA-producing strain and following multiple expression optimizations of the linker between BPLO and ATR1, the BA titer reached 77.53 mg/L in shake flasks and subsequently reached 205.74 mg/L via fed-batch fermentation in a 5-L bioreactor. In this study, we developed a feasible approach for the de novo synthesis of BA and its direct precursor lupeol in engineered S. cerevisiae .
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ACS Applied Nano Materials
·
2024
·
IF 5.3
Cailin Qiao,Xinrou Wang,Yuwei Gao,Yi Ma,Huibo Luo,Suyi Zhang,Danqun Huo,Changjun Hou
A method was proposed to enhance the growth of Saccharomyces cerevisiae cells and increase the alcohol yield. This strategy utilized nanozymes, specifically FeCo-Prussian blue analogue nanoparticles (FeCo-PBA NPs), FeCo-N-C, CoSO porous nanocubes (CoSO PNCs), and FeN3P-single-atom nanozymes (FeN3P-SAzyme), to mimic the antioxidant enzymes. When subjected to H2O2 stress, the nanozymes significantly stimulated cell growth and effectively reduced the levels of reactive oxygen species (ROS), as evidenced by the decrease in the malondialdehyde (MDA) content. Additionally, the decline in lactate dehydrogenase (LDH) activity within the fermentation environment suggested that nanozymes provided protection against cell membrane damage caused by H2O2. This protective effect can be attributed to their inhibitory impact on ROS production. Further results demonstrated that 50 μg mL–1 of nanozymes led to an increase in the ethanol yield and total alcohol yield in S. cerevisiae. This approach held great potential for enhancing alcohol production efficiency in the S. cerevisiae industrial fermentation process.
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LWT-FOOD SCIENCE AND TECHNOLOGY
·
2018
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IF 3.129
Min Li,Weizhu Chen,Zhuo Zhang,Zhiwei Zhang,Bangzhu Peng
Patulin (PAT) is a toxic metabolite produced by several filamentous fungi common to fruit- and vegetable-based products, which results in a potential food safety hazard . Over the past few years, there have been reports stating that PAT was removed by yeast from some fermented products. However, the interactions between PAT and yeast during fermentation are still unclear. In this paper, we focused on the interactions between PAT and S. cerevisiae CITCC 93161 in aqueous solutions during fermentation. The results demonstrated that PAT affected the yeast cells in a time- and concentration-dependent manner, but the yeast exhibited a very strong tolerance to the toxicity of PAT. The yeast cells could completely degraded PAT through the exo- and endo-enzymes synthesized during yeast fermentation, of which the latter exhibited higher activity. Moreover, the PAT biodegradation ability of yeast was not induced by the addition of such toxics to the culture media. Additionally, E-ascladiol was tentatively identified as the product of PAT biodegradation.
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Fermentation-Basel
·
2024
·Vol. 10
·期 4
·211
·
IF 3.3
Qiu Tao,Guocheng Du,Jian Chen,Juan Zhang,Zheng Peng
Patchoulol is a natural sesquiterpene alcohol with extensive applications in cosmetics and pharmaceuticals. In this study, we first constructed the synthesis pathway of patchoulol inSaccharomyces cerevisiaeby expressing the patchoulol synthasePTSgene using the strong promoterGAL1. Afterward, the metabolic flux of the precursor was enhanced by strengthening the mevalonate pathway and balancing the precursor competition pathway, resulting in a 32.74-fold increase in patchoulol production. Subsequently, the supply of acetyl-CoA in yeast was increased by modifying transcriptional regulators and modulating the acetyl-CoA pathway, and the titer of patchoulol reached 155.94 mg/L. Finally, optimization of the fermentation conditions resulted in a titer of 195.96 mg/L in the shake flasks. Further, batch-fed fermentation in a 5 L bioreactor yielded 1.95 g/L. This work accelerated the development of a microbial cell factory for the production of patchoulol.
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AMB Express
·
2024
·Vol. 14
·期 1
·1-10
·
IF 3.5
Wang Laiyou,Liu Bingbing,Meng Qingshan,Yang Chunchun,Hu Yiyi,Wang Chunyan,Wu Pengyu,Ruan Chen,Li Wenhuan,Cheng Shuang,Guo Shuxian
The unsustainable and widespread utilization of fossil fuels continues to drive the rapid depletion of global supplies. Biodiesel has emerged as one of the most promising alternatives to conventional diesel, leading to growing research interest in its production. Microbes can facilitate the de novo synthesis of a type of biodiesel in the form of fatty acid methyl esters (FAMEs). In this study, Saccharomyces cerevisiae metabolic activity was engineered to facilitate enhanced FAME production. Initially, free fatty acid concentrations were increased by deleting two acetyl-CoA synthetase genes (FAA1, FAA4) and an acyl-CoA oxidase gene (POX1). Intracellular S-adenosylmethionine (SAM) levels were then enhanced via the deletion of an adenosine kinase gene (ADO1) and the overexpression of a SAM synthetase gene (SAM2). Lastly, the S. cerevisiae strain overproducing free fatty acids and SAM were manipulated to express a plasmid encoding the Drosophila melanogaster Juvenile Hormone Acid O-Methyltransferase (DmJHAMT). Using this combination of engineering approaches, a FAME concentration of 5.79 ± 0.56 mg/L was achieved using these cells in the context of shaking flask fermentation. To the best of our knowledge, this is the first detailed study of FAME production in S. cerevisiae. These results will provide a valuable basis for future efforts to engineer S. cerevisiae strains for highly efficient production of biodiesel.
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Food Bioscience
·
2026
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IF 6.2
Zhiqiang Yi,Weizhong Hu,Huiying Li,Shuxin Zhou,Jing Chengjun,Quang D. Nguyen,Szilárd Kun,Szandra Klátyik,Zhirong Wang,Jianquan Kan,Muying Du
In this study, four Saccharomyces cerevisiae strains were evaluated for their fermentation performance and the resulting wine’s quality. The results demonstrated that the Saccharomyces cerevisiae SY exhibited the most robust fermentation capacity, with faster initiation and higher efficiency. The SY group exhibited a higher total content of active ingredients (1.04 mg/mL), with Parishin E levels being significantly higher compared to the others. Although the amino acid content of PM group was found to be the highest (63.36 mg/L), while the percentage of bitter amino acid in the SY group was found to be the lowest (39.52%). and the percentage of umami amino acid in the SY group was found to be the highest (28.1%). The DV10 group exhibited the highest levels of pyruvic and malic acid (0.53 g/L and 1.64 g/L, respectively) while the SY group demonstrated the lowest acetic acid content (1.63 g/L). In addition, the SY group had a more complex aroma composition, with higher scores for sensory floral and fruity aromas. Furthermore, Gastrodia elata fermentation wine could reduce the damage of nerve cells in sleep-disordered zebrafish, reduce the distance and time of zebrafish's nocturnal movement, which had effect of improving sleep in zebra fish model, Specific mechanisms of action can be further investigated in the future.
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FOOD CHEMISTRY
·
2026
·
IF 10.4
Shen Yao,Yang Li,Xianwei Zeng,Yuan Cui,Zhaolian Li,Meichen Liu,Daqing Zhao,Shiting Yu,Siming Wang
Fermentation affects ginseng wine flavors. In this study, the kinetic profiles of volatile and non-volatile flavor compounds generated during the fermentation of ginseng wine were monitored by two-dimensional gas chromatography-time-of-flight mass spectrometry and liquid chromatography–mass spectrometry. A sensory analysis revealed that the sample fermented for 7 days had the best flavor balance (prominent fruity aroma and minor herbal notes). Five key flavor compounds were identified: isoamyl alcohol, 2-methylbutyraldehyde, isobutyl acetate, 2-nonenal, and heptanal. Notably, 2-nonenal and heptanal, which are unique ginseng flavor compounds, produced a distinct fatty–fruity composite aroma during fermentation. Correlation analyses indicated that organic acids promoted the production of flavor compounds in ginseng wine and that amino acids served as flavor compound precursors. Sensory reconstruction experiments using exogenous flavor compounds showed that these five flavor compounds significantly affected ginseng wine aromas, especially fatty, fruity, and sweet aromas. These results guide industrial production of high-quality, flavorful, low-alcohol ginseng wine.
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Biotechnology Journal
·
2023
·
IF 4.7
Haiquan Yang,Kunjie Zhang,Wei Shen,Yuanyuan Xia,Yiting Li,Xianzhong Chen
Cembratriene-ol is a good biodegradable biopesticide ingredient with future potential applications in the field of sustainable agriculture. Cembratriene-ol is a monocyclic diterpenoid compound that is synthesized only in the trichome gland of Nicotiana plants. In this study, geranylgeranyl diphosphate synthase gene ggpps from Taxus canadensis and cbts*Δp were heterologously expressed in Saccharomyces cerevisiae W303-1A to successfully synthesize cembratriene-ol. The titer of cembratriene-ol was increased by 1.84-fold compared to the control by overexpressing the S. cerevisiae bifunctional (2E,6E)-farnesyl diphosphate synthase genes ERG20 and cbts*Δp under one promoter P GAP . The titer of cembratriene-ol in the engineered S. cerevisiae BY4741 was increased by 1.39-fold compared to the engineered S. cerevisiae W303-1A. The titer of cembratriene-ol in the engineered S. cerevisiae BY4741 was increased by 2.22-fold compared to the control by overexpressing ERG20 and cbts*Δp , respectively, using two promoters P GAP . Cembratriene-ol was found to be successfully synthesized via the integrated expression of cbts*Δp , ggpps and ERG20 on the genome of S. cerevisiae BY4741. The titer of cembratriene-ol in S. cerevisiae S25 was further increased by 1.80-fold compared to the control via dynamic control of the squalene synthase gene ERG9 . Overexpression of the genes cbts*Δp and ggpps using pY26-GPD-TEF in S. cerevisiae S25 with their integration expression increased the titer of cembratriene-ol by 26.1-fold compared to S. cerevisiae S25. The titer of cembratriene-ol was significantly enhanced by mitochondrial compartmentalized expression of cbts*Δp and ggpps , which was 76.3-fold higher than that of the initial strain constructed. It was indicated that the systematic optimization has great potential in facilitating high-level production of cembratriene-ol production in S. cerevisiae . This article is protected by copyright. All rights reserved
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INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
·
2023
·Vol. 24
·期 10
·9033
·
IF 5.6
Weilan Piao,Chong Li,Pengkun Sun,Miaomiao Yang,Yansong Ding,Wei Song,Yunxiao Jia,Liqun Yu,Yanming Lu,Hua Jin
As a master regulator in cells, RNA-binding protein (RBP) plays critical roles in organismal development, metabolism and various diseases. It regulates gene expression at various levels mostly by specific recognition of target RNA. The traditional CLIP-seq method to detect transcriptome-wide RNA targets of RBP is less efficient in yeast due to the low UV transmissivity of their cell walls. Here, we established an efficient HyperTRIBE (Targets of RNA-binding proteins Identified By Editing) in yeast, by fusing an RBP to the hyper-active catalytic domain of human RNA editing enzyme ADAR2 and expressing the fusion protein in yeast cells. The target transcripts of RBP were marked with new RNA editing events and identified by high-throughput sequencing. We successfully applied HyperTRIBE to identifying the RNA targets of two yeast RBPs, KHD1 and BFR1. The antibody-free HyperTRIBE has competitive advantages including a low background, high sensitivity and reproducibility, as well as a simple library preparation procedure, providing a reliable strategy for RBP target identification inSaccharomyces cerevisiae.Keywords:HyperTRIBE;RNA-binding protein (RBP);yeast;KHD1;BFR1
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Scientific Reports
·
2025
·Vol. 15
·期 1
·1-11
·
IF 3.9
Wahid Ehtisham,Ocheja Ohiemi Benjamin,Oguntomi Sunday Olakunle,Pan Run,Grattieri Matteo,Guaragnella Nicoletta,Guaragnella Cataldo,Marsili Enrico
Electrodes functionalised with weak electroactive microorganisms offer a viable alternative to conventional chemical sensors for detecting priority pollutants in bioremediation processes. Biofilm-based biosensors have been proposed for this purpose. However, biofilm formation and maturation require 24–48 h, and the microstructure and coverage of the electrode surface cannot be controlled, leading to poorly reproducible signal and sensitivity. Alternatively, semiconductive biocompatible coatings can be used for viable cell immobilization, achieving reproducible coverage and resulting in a stable biosensor response. In this work, we use a polydopamine (PDA)-based coating to immobilize Saccharomyces cerevisiae yeast viable cells on carbon screen printed electrodes (SPE) for Cu(II) detection, with potassium ferricyanide (K 3 [Fe (CN) 6 ]) as a redox mediator. Under these conditions, the current output correlates with Cu (II) concentration, reaching a limit of detection of 2.2 µM, as calculated from the chronoamperometric response. The bioelectrochemical results are supported by standard viability assays, microscopy, and electrochemical impedance spectroscopy. The PDA coatings can be functionalised with different mutant strains, thus expanding the toolbox for biosensor design in bioremediation.
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BIOCHEMICAL ENGINEERING JOURNAL
·
2025
·
IF 3.8
Junjie Wang,Pengming Yang,Shuai Tu,Weihong Zhong
Scopoletin, a plant-derived coumarin, exhibits diverse pharmacological activities. Nowadays, scopoletin is mainly produced by plant extraction. In this study, a Saccharomyces cerevisiae strain capable of de novo scopoletin biosynthesis was constructed by successively integrating ten heterologous genes into a p -coumaric acid producing strain. The following strategies were employed to improve the yield of scopoletin: (1) A high-yield caffeic acid platform strain (301.98 mg/L) was established by increasing the HpaBC copy number, integration of EcaroL and RgTAL , feedback-resistant mutant expression cassette ( ARO3 D154N / ARO4 G141S / ARO7 K229L ), and knockout competing pathway genes ( ARO10 / PDC5 ); (2) Two distinct pathways from caffeic acid to scopoletin were introduced and selected; (3) Integration of heterologous genes AtHCT and AtC3’H , to construct the pathway from p -coumaroyl-CoA to caffeoyl-CoA; (4) Attenuating PYK1 by introducing the mutant gene of PYK1 D146N to increase PEP supply; (5) Overexpression of cofactor regeneration genes ( ADO1 , SAH1 , and ZWF1 ) to strengthen the supply of SAM and NADPH; (6) Adjustment of the fermentation medium. The engineered strain produced scopoletin of 55.32 mg/L in shake flasks. To our knowledge, this is the first report of de novo scopoletin biosynthesis in S. cerevisiae . These results establish new insights for microbial production of scopoletin and coumarin derivatives.
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ACS Synthetic Biology
·
2024
·
IF 3.7
Mei Tang,Xianhao Xu,Yanfeng Liu,Jianghua Li,Guocheng Du,Xueqin Lv,Long Liu
Betulinic acid (BA) is a lupane-type triterpenoid with potent anticancer and anti-HIV activities. Its great potential in clinical applications necessitates the development of an efficient strategy for BA synthesis. This study attempted to achieve efficient BA biosynthesis in Saccharomyces cerevisiae using systematic metabolic engineering strategies. First, a de novo BA biosynthesis pathway in S. cerevisiae was constructed, which yielded a titer of 14.01 ± 0.21 mg/L. Then, by enhancing the BA synthesis pathway and dynamic inhibition of the competitive pathway, a greater proportion of the metabolic flow was directed toward BA synthesis, achieving a titer of 88.07 ± 5.83 mg/L. Next, acetyl-CoA and NADPH supply was enhanced, which increased the BA titer to 166.43 ± 1.83 mg/L. Finally, another BA synthesis pathway in the peroxisome was constructed. Dual regulation of the peroxisome and cytoplasmic metabolism increased the BA titer to 210.88 ± 4.76 mg/L. Following fed-batch fermentation process modification, the BA titer reached 682.29 ± 8.16 mg/L. Overall, this work offers a guide for building microbial cell factories that are capable of producing terpenoids with efficiency.
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