BIOTECHNOLOGY AND BIOENGINEERING
·
2020
·Vol. 117
·期 8
·2410-2419
·
IF 4.002
Wei Guo,Qiulan Huang,Yuhui Feng,Taicong Tan,Suhao Niu,Shaoli Hou,Zhigang Chen,Zhi-Qiang Du,Yu Shen,Xu Fang
Metabolic engineering of Saccharomyces cerevisiae for high-level production of aromatic chemicals has received increasing attention in recent years. Tyrosol production from glucose by S. cerevisiae is considered an environmentally sustainable and safe approach. However, the production of tyrosol and salidroside by engineered S. cerevisiae has been reported to be lower than 2 g/L to date. In this study, S. cerevisiae was engineered with a push-pull-restrain strategy to efficiently produce tyrosol and salidroside from glucose. The biosynthetic pathways of ethanol, phenylalanine, and tryptophan were restrained by disrupting PDC1 , PHA2 , and TRP3 . Subsequently, tyrosol biosynthesis was enhanced with a metabolic pull strategy of introducing PcAAS and EcTyrA M53I/A354V . Moreover, a metabolic push strategy was implemented with the heterologous expression of phosphoketolase (Xfpk), and then erythrose 4-phosphate was synthesized simultaneously by two pathways, the Xfpk-based pathway and the pentose phosphate pathway, in S. cerevisiae . Furthermore, the heterologous expression of Xfpk alone in S. cerevisiae efficiently improved tyrosol production compared with the coexpression of Xfpk and phosphotransacetylase. Finally, the tyrosol yield increased by approximately 135-folds, compared with that of parent strain. The total amount of tyrosol and salidroside with glucose fed-batch fermentation was over 10 g/L and reached levels suitable for large-scale production.
研究领域:
生物技术
·
代谢工程
✓ 已验证
DOI
PubMed
高影响因子
BIOCHEMICAL ENGINEERING JOURNAL
·
2025
·
IF 3.8
Jiayan Du,Youtong Bao,Jingyuan Zhu,Xueqing Pang,Depeng Ren,Xinjian Yin,Pingping Zhou
Astaxanthin, a potent antioxidant widely applied in health supplements, cosmetics, and animal feed, faces production challenges in microbial synthesis due to inefficient β-carotene hydroxylase activity. This study identified rate-limiting enzymes and demonstrated that the expressing phytoene desaturase BtCrtI from Blakeslea trispora in Saccharomyces cerevisiae increased β-carotene production 2.8-fold compared to XdCrtI from Xanthophyllomyces dendrorhous . Integrating of XdCrtYB, BtCrtI, CrtE03M, and tHMG1 achieved 208.7 mg/L β-carotene. However, introducing β-carotene ketolase and β-carotene hydroxylase into this strain unexpectedly increased lycopene accumulation, likely due to feedback inhibition by the product and enzyme imbalance. To address this, we utilized a moderate β-carotene producer YJYast-02 for astaxanthin production. Truncating the N-terminal 39 amino acids of β-carotene hydroxylase from Haematococcus pluvialis improved astaxanthin titer by 37.7 %. Rational engineering of the hydrophobicity of the substrate-binding pocket in β-carotene hydroxylase, specifically through mutations such as N183A, C191A and T213A, further increased astaxanthin titer by 15.8 %, 30.5 %, 36.8 %, respectively. The final engineered strain, YJYast-03(t5t39 T213A ), expressing t5OBKTM30 and t39OcrtZM1 T213A , achieved 26.0 mg/L (3.6 mg/g DCW) astaxanthin. This work pioneers the use of molecular docking for engineering β-carotene hydroxylase, offering potential to significantly enhance catalytic activity and optimize astaxanthin biosynthesis in yeast.
研究领域:
生物工程
·
合成生物学
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微生物学
✓ 已验证
产品关联
DOI
高影响因子
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
·
2020
·
IF 4.192
Hong Sun,Jingli Yang,Xue Lin,Congfa Li,Yongjin He,Zhigang Cai,Guoyin Zhang,Hao Song
Delta-tocotrienol as a vitamin E isomer has received much attention because of its diverse biomedical applications. Microbial biosynthesis of delta-tocotrienol is a promising strategy for its economic and environmental advantages. Here, we accomplished complete biosynthesis of delta-tocotrienol in Saccharomyces cerevisiae from glucose. We first constructed and incorporated a heterologous pathway into the genome of S. cerevisiae by incorporating the genes hpd (from Pseudomonas putida KT2440), hpt (from Synechocystis sp. PCC 6803), and vte1 (from Arabidopsis thaliana) for the biosynthesis of delta-tocotrienol. We further enhanced the biosynthesis of the precursor geranylgeranyl diphosphate by overexpressing the thmg1 and ggppssa (from Sulfolobus acidocaldarius) genes, leading to a production titer of delta-tocotrienol of 1.39 ± 0.01 mg/L. Finally, we optimized the fermentation medium using the response surface methodology, enabling a high-titer production of delta-tocotrienol (3.56 ± 0.25 mg/L), ∼2.6-fold of that of the initial culture medium. Fed-batch fermentation in a 2 L fermenter was further used to enhance the production titer of delta-tocotrienol (4.10 ± 0.10 mg/L). To the best of our knowledge, this is the first report on the de novo biosynthesis of delta-tocotrienol in S. cerevisiae, and the highest titer obtained for microbial production of delta-tocotrienol.
研究领域:
生物技术
·
微生物学
·
代谢工程
✓ 已验证
DOI
PubMed
高影响因子
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
·
2025
·
IF 6.2
Yuan Zhu,Xiaoguang Yan,Weiguo Li,Jianjun Qiao,Guang-Rong Zhao
Ursolic acid, a plant-derived pentacyclic triterpenoid with anti-inflammatory, antioxidant, and other bioactive properties, holds significant potential for use in nutritional supplements and drug development. However, its extraction from medicinal plants is inefficient due to low yield and dependence on seasonality and geography. Herein, we use modular metabolic engineering to enhance ursolic acid production in Saccharomyces cerevisiae by dividing the biosynthetic pathway into five modules. First, the heterologous ursolic acid biosynthesis module was established using Catharanthus roseus α-amyrin synthase (CrMAS) and a fused α-amyrin oxidase (CrOAS) with cytochrome P450 reductase (CPR). Next, the full hybrid mevalonate pathway was overexpressed, and the copy number of CrMAS was optimized. The sterol pathway was further optimized by introducing N-degron tags to relieve the competition pathway and deleting the SSM4 gene to enhance the ERG1 stability. Acetyl-CoA supply was improved via phosphoketolase and acetyl-CoA synthase pathways, combined with fine-tuning of mitochondrial and cytosolic carbon flux. The final engineered strain produced 1083.62 mg/L of ursolic acid in shake-flask cultures and 8.59 g/L in a 5 L bioreactor via fed-batch fermentation, achieving the highest microbial ursolic acid titer reported to date. This study not only demonstrates the potential for efficient biosynthesis of triterpenoid compounds but also provides ideas that can be extended to other microbial hosts for the concentrated use of intracellular carbon sources in the synthesis of complex natural products.
研究领域:
代谢工程
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生物技术
·
生物化学
·
微生物生物技术
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Collagen and Leather
·
2025
·Vol. 7
·期 1
·1-14
·
IF 9.2
Yang Ming,Li Xia,Wang Bo,Liu Xian,Zhang Bo,Liao Xue-Pin
The efficient biosynthesis is important for the sustainable development of lignocellulosic ethanol industry, but it is limited by furfural stress produced with cellulose pretreatment. Collagen peptide (CP), as an affluent protein resource, considerably improved the tolerance of Saccharomyces cerevisiae against furfural stress. When the furfural concentration was 2 g/L, the residual sugar concentration was reduced from 122.39 to 8.90 g/L, and the final ethanol yield increased from 30.69 to 87.27 g/L in the presence of CP. In addition, the ethanol yield in CP containing media was higher than those in other peptides. Transcriptome analysis showed CP can improve the expression of genes ( FBA1 , PDC1 , PDC6 , and ENO1 ) associated with glycolysis to promote sugar utilization, and enhance ethanol biosynthesis under furfural stress, which were further verified by quantitative real-time PCR. These results indicated that CP is a promising protectant and accelerator for bioethanol biosynthesis. Graphical abstract
研究领域:
微生物学
·
生物工程
✓ 已验证
产品关联
DOI
高影响因子
Environmental Technology & Innovation
·
2025
·
IF 7.1
Yinhua Cui,Yiwen Chen,Qingshan Shi,Jin Feng,Yafeng Song,Junwei Yao,Xiaobao Xie
A growing number of microorganisms are employed in industry nowadays, leading to the extensive generation of microbial waste. Nevertheless, the waste recycling investigation has not yet encompassed the synthesis of nanoparticles. Economically viable nano-selenium (nano-Se) requires additional cost reduction. This study produced cost-effective nano-Se utilizing microbial waste. This reaction was conducted under UV irradiation due to the low activity of microbial waste. After 90 min of UV irradiation, the yield of nano-Se generated from the death-phase eukaryote Saccharomyces cerevisiae ( S. cerevisiae ) waste was 7.46 times more than that of nano-Se produced in darkness. The resultant nano-Se is amorphous, and the nano-Se generated by UV irradiation demostrated a larger size than that produced in darkness. Moreover, unlike the nano-Se generated in darkness or incubators, the nano-Se produced under UV irradiation significantly inhibited the multiplication of MDA-MB-231 cancer cells, demonstrating its enhanced efficacy. The UV-assisted synthesis of nano-Se was also seen during the death phase of the prokaryote Escherichia coli ( E. coli ) waste. After 5 h of UV irradiation, the yield of nano-Se produced by E. coli is 10.80 times more than that produced in darkness. The findings strongly indicate that industrial microbial waste, whether eukaryote or prokaryote, can be employed for the economical production of nano-Se or other nanoparticles.
✓ 已验证
产品关联
DOI
高影响因子
Food & Function
·
2023
·Vol. 14
·期 7
·3083-3091
·
IF 6.1
Ruiyu Zhu,Shuo Shan,Su Zhou,Zhen Chen,Yuanfeng Wu,Wei Liao,Chao Zhao,Qiang Chu
Patulin is one of the mycotoxins that exists in abundance in fruits and derivative products and is easily exposed in daily life, leading to various toxicities such as genotoxicity, teratogenicity, immunotoxicity, and carcinogenicity in the human body, while the efficient removal or degradation measures are still in urgent demand. In this work, Saccharomyces cerevisiae, a natural yeast with both patulin degradation and intestine damage protection abilities, was first applied to prevent and decrease the hazard after patulin intake. In vitro, Saccharomyces cerevisiae KD (S. cerevisiae KD) could efficiently degrade patulin at high concentrations. In a Canenorhabditis elegans (C. elegans) model fed on S. cerevisiae KD, locomotion, oxidative stress, patulin residual, intestine damage, and gene expression were investigated after exposure to 50 μg mL−1 patulin. The results demonstrated that S. cerevisiae KD could efficiently degrade patulin, as well as weaken the oxidative stress and intestinal damage caused by patulin. Moreover, S. cerevisiae KD could regulate the gene expression levels of daf-2 and daf-16 through the IGF-1 signaling pathway to control the ROS level and glutathione (GSH) content, thus decreasing intestinal damage. In summary, this work uncovers the outstanding characteristic of an edible probiotic S. cerevisiae KD in patulin degradation and biotoxicity alleviation and provides enlightenment toward solving the hazards caused by the accumulation of patulin.
研究领域:
食品科学
·
毒理学
·
微生物学
·
分子生物学
✓ 已验证
DOI
PubMed
高影响因子
Autophagy
·
2021
·
IF 16.016
Di Peng,Chen Ruan,Shanshan Fu,Chengwen He,Jingzhen Song,Hui Li,Yiran Tu,Dachao Tang,Lan Yao,Shaofeng Lin,Ying Shi,Weizhi Zhang,Hao Zhou,Le Zhu,Cong Ma,Cheng Chang,Jie Ma,Zhiping Xie,Chenwei Wang,Yu Xue
In Saccharomyces cerevisiae, Atg9 is an important autophagy-related (Atg) protein, and interacts with hundreds of other proteins. How many Atg9-interacting proteins are involved in macroautophagy/autophagy is unclear. Here, we conducted a multi-omic profiling of Atg9-dependent molecular landscapes during nitrogen starvation-induced autophagy, and identified 290 and 256 genes to be markedly regulated by ATG9 in transcriptional and translational levels, respectively. Unexpectedly, we found most of known Atg proteins and autophagy regulators that interact with Atg9 were not significantly changed in the mRNA or protein level during autophagy. Based on a hypothesis that proteins with similar molecular characteristics might have similar functions, we developed a new method named inference of functional interacting partners (iFIP) to integrate the transcriptomic, proteomic and interactomic data, and predicted 42 Atg9-interacting proteins to be potentially involved in autophagy, including 15 known Atg proteins or autophagy regulators. We validated 2 Atg9-interacting partners, Glo3 and Scs7, to be functional in both bulk and selective autophagy. The mRNA and protein expressions but not subcellular localizations of Glo3 and Scs7 were affected with or without ATG9 during autophagy, whereas the colocalizations of the 2 proteins and Atg9 were markedly enhanced at early stages of the autophagic process. Further analyses demonstrated that Glo3 but not Scs7 regulates the retrograde transport of Atg9 during autophagy. A working model was illustrated to highlight the importance of the Atg9 interactome. Taken together, our study not only provided a powerful method for analyzing the multi-omics data, but also revealed 2 new players that regulate autophagy.Abbreviations: ALP: alkaline phosphatase; Arf1: ADP-ribosylation factor 1; Atg: autophagy-related; Co-IP: co-immunoprecipitation; Cvt: cytoplasm-to-vacuole targeting; DEM: differentially expressed mRNA; DEP: differentially expressed protein; DIC: differential interference contrast; E-ratio: enrichment ratio; ER: endoplasmic reticulum; ES: enrichment score; FC: fold change; FPKM: fragments per kilobase of exon per million fragments mapped; GAP: GTPase-activating protein; GFP: green fluorescent protein; GO: gene ontology; GSEA: gene set enrichment analysis; GST: glutathione S-transferase; HA: hemagglutinin; iFIP: inference of functional interacting partners; KO: knockout; LR: logistic regression; OE: over-expression; PAS: phagophore assembly site; PPI: protein-protein interaction; RFP: red fluorescence protein; RNA-seq: RNA sequencing; RT-PCR: real-time polymerase chain reaction; SCC: Spearman’s correlation coefficient; SD-N: synthetic minimal medium lacking nitrogen; THANATOS: The Autophagy, Necrosis, ApopTosis OrchestratorS; Vsn: variance stabilization normalization; WT: wild-type.
研究领域:
分子生物学
·
基因组学
·
系统生物学
✓ 已验证
DOI
PubMed
高影响因子
JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES
·
2015
·
IF 2.729
Pan Zhu,Song Wang,Jian Wang,Li Zhou,Ping Shi
Adenosine triphosphate and its metabolites are involved in the cellular metabolism process in Saccharomyces cerevisiae. It is very important to simultaneously determine the relative contents of ATP and its metabolites in yeast. In this study, an effective capillary zone electrophoresis method with high selectivity was established. The calibration curves were linear in the concentration range from 1 to 20 mg/L (ATP and cAMP) and 2 to 40 mg/L (ADP and AMP) with excellent correlation coefficients ( r 2 ) > 0.999. The recovery of ATP, ADP, AMP, and cAMP were 99.4%, 94.7%, 100.3% and 99.6%, respectively. Simple sample preparation and easy detection of ATP and its metabolites make this method suitable for the study of changes in the four adenine nucleotides levels caused by caloric restriction in yeast. It is expected that the current method may contribute to further energy metabolism and related investigations of yeast.
研究领域:
生物化学
·
分析化学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Food Bioscience
·
2025
·
IF 5.9
Ruosi Fang,Ting Xia,Shuhui Wu,Jiamin He,Gongnian Xiao
Ethanol produced during the Huangjiu fermentation can adversely affect the growth and metabolism of Saccharomyces cerevisiae . It has been established that arginine, one of the major amino acids found in Huangjiu , can protect yeast cells against ethanol stress. Previous studies have indicated that BTN2 , which influences arginine metabolism, is associated with ethanol tolerance in yeast. In this study, we evaluated the response of BTN2 to ethanol stress based on analyses of cell growth, morphology, membrane permeability, membrane integrity, ROS and how BTN2 influenced intracellular metabolites, enzymes, and transcriptional genes in the arginine metabolic pathway. Our findings revealed that BTN2 promoted significantly increases in the growth and ethanol tolerance of S. cerevisiae g. Under 10% ethanol stress, RT-qPCR showed that BTN2 Δ ( BTN2 knock out strain) up-regulated CAR1 , CAR2 , GAP1 , DUR1,2 and DUR3 to increase arginine and urea degradation, BTN2 ↑ ( BTN2 over-expressing strain) up-regulated CAR1 , CAN1 and DUR3 to enhance arginine uptake, catabolism and urea reabsorption, while TPS1 was up-regulated to improve ethanol tolerance ability. Yeast up-regulated both CAR1 (Arginase) and DUR3 (urea transporter) genes in BTN2 knockout and over-expression strains to counteract the ethanol effects. Our findings revealed a novel arginine metabolism under ethanol tolerance of a core functional yeast during Huangjiu fermentation. These findings could contribute to the development of an approach for inhibiting toxic ethyl carbamate (EC) production during Huangjiu fermentation.
研究领域:
微生物学
·
发酵工程
·
分子生物学
✓ 已验证
产品关联
DOI
高影响因子
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
·
2020
·
IF 4.192
Yun Ma,Zhi Ma,Xiyue Huo,Meiyu Gu,Sainan Ma,Yu Jing,Yixiao Wang,Ying Yue,Zhibiao Feng,Bo Tian
The estimation of yeast viability with B- and N-doped carbon dots (BN-CDs) was investigated in this paper. BN-CDs with a fluorescent quantum yield of 65.47% were prepared by a one-step hydrothermal method. The size distribution of BN-CDs was relatively narrow, with the majority falling within 7.5–8.5 nm, and they were mainly composed of carbon, oxygen, nitrogen, and boron. BN-CDs were shown to have strong and stable fluorescence. They exhibited excitation-independent photoluminescence property, which could avoid the autofluorescence and limitation of the excitation source. Dead and live yeast cells were distinguished well by BN-CD staining in a short time, and there was no strict requirement for light protection. The application of BN-CDs in beer brewing can solve the problem of estimation of yeast viability.
研究领域:
微生物学
·
材料科学
·
生物技术
✓ 已验证
DOI
PubMed
高影响因子
Frontiers in Cell and Developmental Biology
·
2025
·
IF 4.3
Cheng Han,Shanshan Fu,Dachao Tang,Yuting Chen,Dan Liu,Zihao Feng,Yujie Gou,Chi Zhang,Weizhi Zhang,Leming Xiao,Jiayi Zhang,Cong Yi,Yu Xue,Di Peng
In Saccharomyces cerevisiae , Atg1 is a core autophagy-related (Atg) protein kinase (PK) in regulating macroautophagy/autophagy, by physically interacting with numerous other proteins, or by phosphorylating various substrates. It is unclear how many Atg1-interacting partners and substrates are also involved in regulating autophagy. Here, we conducted transcriptomic, proteomic and phosphoproteomic profiling of Atg1-dependent molecular landscapes during nitrogen starvation-triggered autophagy, and detected 244, 245 and 217 genes to be affected by ATG1 in the autophagic process at mRNA, protein, and phosphorylation levels, respectively. Based on the Atg1 interactome, we developed a novel artificial intelligence (AI) framework, inference of autophagy regulators from multi-omic data (iAMD), and predicted 12 Atg1-interacting partners and 17 substrates to be potentially functional in autophagy. Further experiments validated that Rgd1 and Whi5 are required for bulk autophagy, as well as physical interactions and co-localizations with Atg1 during autophagy. In particular, we demonstrated that 2 phosphorylation sites (p-sites), pS78 and pS149 of Whi5, are phosphorylated by Atg1 to regulate the formation of Atg1 puncta during autophagy initiation. A working model was illustrated to emphasize the importance of the Atg1-centered network in yeast autophagy. In addition, iAMD was extended to accurately predict Atg proteins and autophagy regulators from other PK interactomes, indicating a high transferability of the method. Taken together, we not only revealed new autophagy regulators from the Atg1 interactome, but also provided a useful resource for further analysis of yeast autophagy.
研究领域:
分子生物学
·
生物信息学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）