MOLECULAR MICROBIOLOGY
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2021
·Vol. 116
·期 3
·783-793
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IF 3.501
Yidong Zhou,Siyuan Gao,Mingliang Zhang,Wankui Jiang,Zhijian Ke,Jiguo Qiu,Jianhong Xu,Qing Hong
Salicylate is a typical aromatic compound widely distributed in nature. Microbial degradation of salicylate has been well studied and salicylate hydroxylases play essential roles in linking the peripheral and ring-cleavage catabolic pathways. The direct hydroxylation of salicylate catalyzed by salicylate-1-hydroxylase or salicylate-5-hydroxylase has been well studied. However, the CoA mediated salicylate 5-hydroxylation pathway has not been characterized in detail. Here, we elucidate the molecular mechanism of the reaction in the conversion of salicylate to gentisate in the carbaryl-degrading strain Rhizobium sp. X9. Three enzymes (salicylyl-CoA ligase CehG, salicylyl-CoA hydroxylase CehH and gentisyl-CoA thioesterase CehI) catalyzed the conversion of salicylate to gentisate via a route, including CoA thioester formation, hydroxylation and thioester hydrolysis. Further analysis indicated that genes cehGHI are also distributed in other bacteria from terrestrial environment and marine sediments. These genomic evidences highlight the role of this salicylate degradation pathway in the carbon cycle of soil organic compounds and marine sediments. Our findings of this three-step strategy enhanced the current understanding of CoA mediated degradation of salicylate.
研究领域:
微生物学
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环境科学
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生物化学
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DOI
PubMed
高影响因子
Biomolecules
·
2024
·Vol. 14
·期 6
·712
·
IF 4.8
Gehad G. Ibrahim,Madhavi Perera,Saadiah A. Abdulmalek,Jinyong Yan,Yunjun Yan
Resveratrol, a phenylpropanoid compound, exhibits diverse pharmacological properties, making it a valuable candidate for health and disease management. However, the demand for resveratrol exceeds the capacity of plant extraction methods, necessitating alternative production strategies. Microbial synthesis offers several advantages over plant-based approaches and presents a promising alternative.Yarrowia lipolyticastands out among microbial hosts due to its safe nature, abundant acetyl-CoA and malonyl-CoA availability, and robust pentose phosphate pathway. This study aimed to engineerY. lipolyticafor resveratrol production. The resveratrol biosynthetic pathway was integrated intoY. lipolyticaby adding genes encoding tyrosine ammonia lyase fromRhodotorula glutinis, 4-coumarate CoA ligase fromNicotiana tabacum, and stilbene synthase fromVitis vinifera. This resulted in the production of 14.3 mg/L resveratrol. A combination of endogenous and exogenous malonyl-CoA biosynthetic modules was introduced to enhance malonyl-CoA availability. This included genes encoding acetyl-CoA carboxylase 2 fromArabidopsis thaliana, malonyl-CoA synthase, and a malonate transporter protein fromBradyrhizobium diazoefficiens. These strategies increased resveratrol production to 51.8 mg/L. The further optimization of fermentation conditions and the utilization of sucrose as an effective carbon source in YP media enhanced the resveratrol concentration to 141 mg/L in flask fermentation. By combining these strategies, we achieved a titer of 400 mg/L resveratrol in a controlled fed-batch bioreactor. These findings demonstrate the efficacy ofY. lipolyticaas a platform for the de novo production of resveratrol and highlight the importance of metabolic engineering, enhancing malonyl-CoA availability, and media optimization for improved resveratrol production.
研究领域:
生物技术
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代谢工程
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微生物学
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药学
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生物化学
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DOI
PubMed
高影响因子
FOOD CHEMISTRY
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2020
·
IF 6.306
Ruiqing Long,Ying Guo,Lianwu Xie,Shuyun Shi,Jinju Xu,Chaoying Tong,Qinlu Lin,Te Li
A new-style white pepper derived dual-emission carbon dots (CDs) with a quantum yield of 10.4% was designed and facile constructed with one-pot solvothermal method. The green emission (520 nm) had an efficient and special “turn-on” fluorescence sensing of coenzyme A (CoA) with the aid of Cu 2+ , while red emission (668 nm) barely changed and worked as reference. In the concentration range (0–150 µM), relative fluorescence intensity ratios (F 520 /F 668 ) showed excellent linear correlation with concentrations of CoA, and detection limit was as low as 8.75 nm. Moreover, the strategy has been successfully applied for label-free detection of CoA in real pig liver samples with good recoveries (93.3–108.0%). Notably, the synthesized CDs had durable fluorescence, low cytotoxicity, and good biocompatibility for cellular imaging, which demonstrated wide and promising applicability for biosensing and bioimaging in the future.
研究领域:
分析化学
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生物传感
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生物医学工程
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DOI
PubMed
高影响因子
PLoS Genetics
·
2025
·Vol. 21
·期 5
·e1011704
·
IF 3.7
Zhiming Liu,Gang Du,Yi Chen,Haiyang Chen
The decline in adult stem cell performance is closely linked to tissue malfunction and the rising incidence of age-related diseases. To investigate the molecular basis of these impairments, our screening strategy identified reduced activity in the pantothenate/coenzyme A (CoA) pathway within aged ISCs. Furthermore, exogenous CoA supplementation restructured ISC metabolic pathways, reversing age-induced hyperproliferation and intestinal dysfunction, and thus extending Drosophila lifespan by curbing excessive iron accumulation in ISCs. These findings uncover a new mechanism of stem cell aging and propose that pantothenate and CoA could be potential therapeutic targets for treating age-related diseases and enhancing healthy aging in humans.
In our study, we explored the connection between the aging of adult stem cells and the rise of age-related diseases, uncovering a significant metabolic link. As stem cells age, a key metabolic pathway involving pantothenate and CoA becomes less active, leading to an accumulation of excess iron, which disrupts cellular function and contributes to the aging process. To counteract this decline, we added pantothenate and CoA to the aging stem cells, restoring their normal function, reducing iron buildup, and ultimately improving the health and lifespan of old fruit flies. This simple intervention not only highlights the potential of pantothenate and CoA as powerful tools to combat the effects of aging but also suggests broader implications for treating age-related diseases and promoting healthy aging in humans. Our findings provide a new perspective on the biology of aging and open up exciting new avenues for therapeutic interventions that could improve the quality of life for older individuals.
研究领域:
细胞生物学
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分子生物学
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衰老研究
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营养学
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DOI
PubMed
高影响因子
Scientific Reports
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2021
·Vol. 11
·期 1
·1-13
·
IF 4.38
Niu Meiyun,Yan Haifeng,Xiong Yuping,Zhang Yueya,Zhang Xinhua,Li Yuan,da Silva Jaime A. Teixeira,Ma Guohua
Sandalwood ( Santalum album L.) is famous for its unique fragrance derived from the essential oil of heartwood, whose major components are santalols. To understand the mechanism underlying the biosynthesis of santalols, in this study, we cloned two related genes involved in the mevalonate pathway in S. album coding for acetyl-CoA C-acetyl transferase (AACT) and 3-hydroxy-3-methyglutary-CoA synthase (HMGS). These genes were characterized and functionally analyzed, and their expression profiles were also assessed. An AACT gene designated as SaAACT (GenBank accession No. MH018694) and a HMGS gene designated as SaHMGS (GenBank accession No. MH018695) were successfully cloned from S. album . The deduced SaAACT and SaHMGS proteins contain 415 and 470 amino acids, and the corresponding size of their open-reading frames is 1538 bp and 1807 bp, respectively. Phylogenetic trees showed that the SaAACT protein had the closest relationship with AACT from Hevea brasiliensis and the SaHMGS proteins had the highest homology with HMGS from Siraitia grosvenorii . Functional complementation of SaAACT and SaHMGS in a mutant yeast strain deficient in these proteins confirmed that SaAACT and SaHMGS cDNA encodes functional SaAACT and SaHMGS that mediate mevalonate biosynthesis in yeast. Tissue-specific expression analysis revealed that both genes were constitutively expressed in all examined tissues (roots, sapwood, heartwood, young leaves, mature leaves and shoots) of S. album , both genes showing highest expression in roots. After S. album seedlings were treated with 100 μM methyl jasmonate, the expression levels of SaAACT and SaHMGS genes increased, suggesting that these genes were responsive to this elicitor. These studies provide insight that would allow further analysis of the role of genes related to the sandalwood mevalonate pathway in the regulation of biosynthesis of sandalwood terpenoids and a deeper understanding of the molecular mechanism of santalol biosynthesis.
研究领域:
植物学
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遗传学
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植物生物化学
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分子生物学
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产品关联
DOI
PubMed
高影响因子
Environmental Chemistry and Ecotoxicology
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2025
·
IF 8.2
Xin Wan,Geyu Liang,Dayong Wang
The 6-PPD quinone (6-PPDQ) is derived from 6-PPD, an antioxidant added in tires. Citric acid cycle plays a pivotal role in biochemical metabolisms. In Caenorhabditis elegans , we examined effect of 6-PPDQ exposure on citric acid cycle and the role of alteration in acetyl CoA and pyruvate contents during this process. In citric acid cycle, contents of intermediates were reduced by 0.1–10 μg/L 6-PPDQ. Meanwhile, expression of key enzyme genes ( cts-1 , idh-2 , dlst-1 , and dld-1 ) for citric acid cycle was decreased by 0.1–10 μg/L 6-PPDQ. After 6-PPDQ exposure, citric acid content was reduced by cts-1 RNAi, and NADH content was reduced by idh-2 , dlst-1 , and dld-1 RNAi. Exposure to 6-PPDQ (0.1–10 μg/L) further decreased acetyl CoA and pyruvate contents, and inhibited expression of enzyme genes ( dlat-1 and dld-1 ) governing acetyl CoA synthesis and enzyme genes ( pyk-1 and pyk-2 ) governing pyruvate synthesis. After 6-PPDQ exposure, dlat-1 and dld-1 RNAi reduced contents of acetyl CoA, citric acid, and NADH, and pyk-1 and pyk-2 RNAi reduced contents of pyruvate, citric acid, and NADH. In 6-PPDQ exposed nematodes, cts-1 , idh-2 , dlst-1 , dld-1 , dlat-1 , pyk-1 , and pyk-2 RNAi further caused susceptibility to 6-PPDQ toxicity. Our results suggested 6-PPDQ exposure risk in resulting in damage on citric acid cycle, which was linked to its toxicity induction.
研究领域:
环境毒理学
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分子生物学
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DOI
高影响因子
Cell Reports
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2024
·
IF 7.5
Xia Li,Shihao Song,Xiaohan Kong,Xiayu Chen,Zhuoxian Zhao,Zizi Lin,Yantao Jia,Yong Zhang,Hai-Bin Luo,Qiao-Ping Wang,Lian-Hui Zhang,Wei Qian,Yinyue Deng
Quorum sensing (QS) is a cell-to-cell communication mechanism mediated by small diffusible signaling molecules. Previous studies showed that RpfR controls Burkholderia cenocepacia virulence as a cis -2-dodecenoic acid (BDSF) QS signal receptor. Here, we report that the fatty acyl-CoA ligase DsfR (BCAM2136), which efficiently catalyzes in vitro synthesis of lauryl-CoA and oleoyl-CoA from lauric acid and oleic acid, respectively, acts as a global transcriptional regulator to control B . cenocepacia virulence by sensing BDSF. We show that BDSF binds to DsfR with high affinity and enhances the binding of DsfR to the promoter DNA regions of target genes. Furthermore, we demonstrate that the homolog of DsfR in B. lata , RS02960, binds to the target gene promoter, and perception of BDSF enhances the binding activity of RS02960. Together, these results provide insights into the evolved unusual functions of DsfR that control bacterial virulence as a response regulator of QS signal.
研究领域:
微生物学
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分子生物学
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产品关联
DOI
PubMed
高影响因子
ECOTOXICOLOGY AND ENVIRONMENTAL SAFETY
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2023
·
IF 6.8
Wenjuan Zhang,Xuerui Ma,Yijia Zhang,Wanjing Tong,Xing Zhang,Yong Liang,Maoyong Song
Bisphenol P (BPP), structurally similar to bisphenol A, is commonly identified in the samples of environment, food, and humans. Unfortunately, very little information is currently available on adverse effects of BPP. The obesogenic effects and underlying mechanisms of BPP on mice were investigated in this study. Compared with the control, high-resolution microcomputed tomography (micro-CT) scans displayed that the visceral fat volume of mice was significantly increased at a dose of 5 mg/kg/day after BPP exposure for 14 days, whereas the subcutaneous fat volume remained unchanged. Nontargeted metabolomic analysis revealed that BPP significantly perturbed the metabolic pathways of mouse livers, and acetyl-CoA was identified as the potential key metabolite responsible for the visceral fat induced by BPP. These findings recommend that a great deal of attention should be paid to the obesogenic properties of BPP as a result of its widely utilized and persistence in the environment.
研究领域:
环境科学
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代谢生物学
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毒理学
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产品关联
DOI
PubMed
高影响因子
Microbial Cell Factories
·
2016
·Vol. 15
·期 1
·1-9
·
IF 3.744
Yang Jianming,Nie Qingjuan
Background Under aerobic conditions, acetic acid is the major byproduct produced by E. coli during the fermentation. And acetic acid is detrimental to cell growth as it destroys transmembrane pH gradients. Hence, how to reduce the production of acetic acid and how to utilize it as a feedstock are of intriguing interest. In this study, we provided an evidence to produce β-caryophyllene by the engineered E. coli using acetic acid as the only carbon source. Results Firstly, to construct the robust acetate-utilizing strain, acetyl-CoA synthases from three different sources were introduced and screened in the E. coli . Secondly, to establish the engineered strains converting acetic acid to β-caryophyllene, acetyl-CoA synthase (ACS), β-caryophyllene synthase (QHS1) and geranyl diphosphate synthase (GPPS2) were co-expressed in the E. coli cells. Thirdly, to further enhance β-caryophyllene production from acetic acid, the heterologous MVA pathway was introduced into the cells. What’s more, acetoacetyl-CoA synthase (AACS) was also expressed in the cells to increase the precursor acetoacetyl-CoA and accordingly resulted in the increase of β-caryophyllene. The final genetically modified strain, YJM67, could accumulate the production of biomass and β-caryophyllene up to 12.6 and 1.05 g/L during 72 h, respectively, with a specific productivity of 1.15 mg h −1 g −1 dry cells, and the conversion efficiency of acetic acid to β-caryophyllene (gram to gram) reached 2.1 %. The yield of β-caryophyllene on acetic acid of this strain also reached approximately 5.6 % of the theoretical yield. Conclusions In the present study, a novel biosynthetic pathway for β-caryophyllene has been investigated by means of conversion of acetic acid to β-caryophyllene using an engineered Escherichia coli . This was the first successful attempt in β-caryophyllene production by E. coli using acetic acid as the only carbon source. Therefore, we have provided a new metabolic engineering tool for β-caryophyllene synthesis.
研究领域:
生物技术
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代谢工程
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微生物学
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DOI
PubMed
高影响因子
PLANT PHYSIOLOGY AND BIOCHEMISTRY
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2025
·
IF 5.7
Minghao Jiang,Xin Chen,Tianqiao He,Wei Hu,Ning Zhao,Min Liao,Haiqun Cao
Chinese sprangletop ( Leptochloa chinensis (L.) Nees) is among the most economically damaging and disruptive weeds found in rice fields across China, and it exhibits a high degree of cyhalofop-butyl resistance for the past few years. Here, a suspected cyhalofop-butyl-resistant population HFCH5 (R) was collected from a rice field and analyzed to explore the mechanistic basis for its resistance activity. Compared with the susceptible HFLY1 (S) population, this R population exhibited differing levels of resistance to four ACCase-inhibiting herbicides (cyhalofop-butyl (RI = 11.52), fenoxaprop- P -ethyl (RI = 11.05), metamifop (RI = 6.61), and clethodim (RI = 4.06)), although it did exhibit susceptibility to florpyrauxifen-benzyl, tripyrasulfone, pyraclonil, and anilofos. This cyhalofop-butyl resistance in R plants was not effectively mitigated by pre-treatment with the cytochrome P450 inhibitor piperonyl butoxide or the glutathione S-transferase inhibitor 4-chloro-7-nitrobenzoxadiazole. Further analyses of these R plants revealed the substitution of Trp-1999 (TGG) for a Gly residue (GGG) within the ACCase carboxyltransferase domain. When the wild-type and mutated forms of this ACCase protein were subjected to structural modeling while complexed with a range of herbicides with ACCase-inhibiting activity, this Trp-1999-Gly substitution was found to be associated with a slight decrease in the affinity of this protein for a series of ACCase-inhibiting herbicides. A dCAPS marker was developed to aid in the rapid detection of this Trp (TGG)-1999-Gly (GGG) mutation within L. chinensis . Notably, before or after four ACCase-inhibiting herbicides (cyhalofop-butyl, fenoxaprop- P -ethyl, metamifop and clethodim) treatment, ACCase gene expression level of the R plants was not significantly higher (fold change <2, P > 0.05) than that of the S plants. This study is the first report that in grass species, the newly identified ACCase Trp-1999-Gly mutation is associated with ACCase-inhibiting herbicide resistance.
研究领域:
植物学
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遗传学
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农业科学
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生物化学
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产品关联
DOI
PubMed
高影响因子
Nature Communications
·
2016
·Vol. 7
·期 1
·1-12
·
IF 11.329
Lv Xiaomei,Wang Fan,Zhou Pingping,Ye Lidan,Xie Wenping,Xu Haoming,Yu Hongwei
Microbial production of isoprene from renewable feedstock is a promising alternative to traditional petroleum-based processes. Currently, efforts to improve isoprenoid production in Saccharomyces cerevisiae mainly focus on cytoplasmic engineering, whereas comprehensive engineering of multiple subcellular compartments is rarely reported. Here, we propose dual metabolic engineering of cytoplasmic and mitochondrial acetyl-CoA utilization to boost isoprene synthesis in S. cerevisiae . This strategy increases isoprene production by 2.1-fold and 1.6-fold relative to the recombinant strains with solely mitochondrial or cytoplasmic engineering, respectively. By combining a modified reiterative recombination system for rapid pathway assembly, a two-phase culture process for dynamic metabolic regulation, and aerobic fed-batch fermentation for sufficient supply of acetyl-coA and carbon, we achieve 2527, mg l −1 of isoprene, which is the highest ever reported in engineered eukaryotes. We propose this strategy as an efficient approach to enhancing isoprene production in yeast, which might open new possibilities for bioproduction of other value-added chemicals.
研究领域:
生物技术
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代谢工程
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微生物学
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产品关联
DOI
PubMed
高影响因子
Cell Discovery
·
2024
·Vol. 10
·期 1
·1-7
·
IF 13
Li Yang,Chen Zhi-Peng,Xu Da,Wang Liang,Cheng Meng-Ting,Zhou Cong-Zhao,Chen Yuxing,Hou Wen-Tao
Human ABC transporters ABCD1–3 are all localized on the peroxisomal membrane and participate in the β-oxidation of fatty acyl-CoAs, but they differ from each other in substrate specificity. The transport of branched-chain fatty acids from cytosol to peroxisome is specifically driven by ABCD3, dysfunction of which causes severe liver diseases such as hepatosplenomegaly. Here we report two cryogenic electron microscopy (cryo-EM) structures of ABCD3 bound to phytanoyl-CoA and ATP at resolutions of 2.9 Å and 3.2 Å, respectively. A pair of phytanoyl-CoA molecules were observed in ABCD3, each binding to one transmembrane domain (TMD), which is distinct from our previously reported structure of ABCD1, where each fatty acyl-CoA molecule strongly crosslinks two TMDs. Upon ATP binding, ABCD3 exhibits a conformation that is open towards the peroxisomal matrix, leaving two extra densities corresponding to two CoA molecules deeply embedded in the translocation cavity. Structural analysis combined with substrate-stimulated ATPase activity assays indicated that the present structures might represent two states of ABCD3 in the transport cycle. These findings advance our understanding of fatty acid oxidation and the molecular pathology of related diseases.
研究领域:
生物化学
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分子生物学
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医学遗传学
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DOI
高影响因子
危险品化学品经营许可证（带存储）