JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
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2024
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IF 5.7
Gege Ma,Xiangyu Zhu,Dongxin Zhang,Haoran Li,Jinping Lin,Dongzhi Wei
(R)-Citronellal is a key chiral precursor of high-value chemicals, such as the best-selling flavor compound (−)-menthol; however, the conventional synthesis suffers from low yield and unsatisfactory enantioselectivity. In this study, we developed a highly atom-efficient hydrogen-borrowing cascade for the synthesis of (R)-citronellal from geraniol using alcohol dehydrogenase from Escherichia coli K12 (AdhP) and ene-reductase from Saccharomyces cerevisiae YJM1341 (OYE2p). The key rate-limiting enzyme, AdhP, was subjected to structure-guided semirational engineering, and the triple mutant AdhP260T/284A/268P (M3) was obtained that demonstrated a 1.28-fold improvement in catalytic efficiency (kcat/Km) toward geraniol. After optimization of the reaction conditions, the hydrogen-borrowing cascade system achieved the conversion of 23.14 g/L geraniol into (R)-citronellal at a conversion rate of 98.23% with 96.7% ee. This work represents an alternative approach for the biosynthesis of (R)-citronellal without sacrificing a cosubstrate or additional enzymes.
研究领域:
生物化学
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合成生物学
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酶学
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DOI
PubMed
高影响因子
Journal of Fungi
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2022
·Vol. 8
·期 12
·1310
·
IF 5.724
Ai-Ai Zhou,Rong-Yu Li,Fei-Xu Mo,Yi Ding,Ruo-Tong Li,Xue Guo,Ke Hu,Ming Li
Background: Natural products are often favored in the study of crop pests and diseases. Previous studies have shown that citronellal has a strong inhibition effect onMagnaporthe oryzae. The objective of this study was to clarify its mechanism of action againstM. oryzae. Results: Firstly, the biological activity of citronellal againstM. oryzaewas determined by direct and indirect methods, and the results show that citronellal had a strong inhibition effect onM. oryzaewith EC50values of 134.00 mg/L and 70.48 μL/L air, respectively. Additionally, a preliminary study on its mechanism of action was studied. After citronellal treatment, electron microscopy revealed that the mycelium became thin and broken; scanning electron microscopy revealed that the mycelium was wrinkled and distorted; and transmission electron microscopy revealed that the mycelium cell wall was invaginated, the mass wall of mycelium was separated, and the organelles were blurred. The mycelium was further stained with CFW, and the nodes were blurred, while the mycelium was almost non-fluorescent after PI staining, and there was no significant difference in the relative conductivity of mycelium. In addition, chitinase was significantly enhanced, and the expression of chitin synthesis-related genes was 17.47-fold upregulated. Finally, we found that the efficacy of citronellal against the rice blast was as high as 82.14% according to indoor efficacy tests. Conclusion: These results indicate that citronellal can affect the synthesis of chitin inM. oryzaeand damage its cell wall, thereby inhibiting the growth of mycelium and effectively protecting rice from rice blasts.Keywords:magnaporthe oryzae;citronellal;cell wall;physiology;cytotoxicity
研究领域:
植物病理学
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真菌学
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农药科学
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PubMed
高影响因子
BIOTECHNOLOGY AND BIOENGINEERING
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2025
·
IF 3.6
Liang Li,Hui Jiang,Tingting Xu,Xi Wang
Citronellal and citronellol are monoterpenoids that serve as key ingredients in the flavor and fragrance industry and hold significant potential for pharmaceutical applications. Currently, these compounds are primarily obtained through chemical synthesis or extraction from plant essential oils. However, these methods raise concerns regarding environmental sustainability, carbon emissions, and the growing attentions for natural production. A promising alternative is microbial biosynthesis, which enables the natural and carbon-neutral production of these compounds in an environmentally friendly manner. In this study, we developed bienzymatic cascades for the efficient production of citronellal and citronellol from geraniol, utilizing E. coli 's endogenous glycerol dehydrogenase (GLDA), NADPH-dependent aldehyde reductase (AHR), and Catharanthus roseus iridoid synthase (CrIS). The highest in vitro production reached 92 mg/L of citronellal and 170 mg/L of citronellol in a 5-h enzymatic reaction. We also engineered these enzymatic cascades into E. coli strains for in vivo production, achieving up to 714 mg/L of citronellol. This study establishes new bienzymatic cascades for the efficient biosynthesis of citronellal and citronellol, demonstrating a possible bio-based production for natural flavor and fragrance compounds.
研究领域:
合成生物学
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生物催化
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代谢工程
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工业微生物学
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酶学
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DOI
PubMed
高影响因子
Journal of Fungi
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2021
·Vol. 7
·期 6
·432
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IF 5.816
Qiuli OuYang,Yangmei Liu,Okwong Reymick Oketch,Miaoling Zhang,Xingfeng Shao,Nengguo Tao
Ergosterol (ERG) is a potential target for the development of antifungal agents againstPenicillium digitatum, the pathogen of green mold in citrus fruits. This study examined the mechanism by which citronellal, a typical terpenoid ofCymbopogon nardusessential oil, acts on ergosterol to exhibit its antifungal activity againstP. digitatum. We previously reported that citronellal inhibited the growth ofP. digitatumwith minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) of 1.36 and 2.72 mg/mL, respectively. In citronellal-treated cells, the membrane integrity and ergosterol contents significantly decreased, whereas lanosterol, which serves as a precursor for ergosterol biosynthesis, massively accumulated. Addition of 150 mg/L of exogenous ergosterol decreased the inhibitory rate of citronellal, restoring the ergosterol content and hence the membrane structure to normal levels, and triggered expression of nearly all ERG genes. Based on our findings, we deduce that citronellal damages the cell membrane integrity ofP. digitatumby down-regulating the ERG genes responsible for conversion of lanosterol to ergosterol, the key downregulated gene beingERG3, due to the observed accumulation of ergosta-7,22-dienol.Keywords:citrus;pathogen;ergosterol;ERG genes;cell membrane integrity
研究领域:
微生物学
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植物病理学
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植物化学
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DOI
PubMed
高影响因子
PHYTOTHERAPY RESEARCH
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2026
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IF 8.1
Yi-wen Qian,Yao Lu,Miao-miao Chang,Qian-qian Niu,Jia-xin Fan,Shi-ying Xing,Peng Li,Ling Liu
Cardiomyocyte senescence contributes to the progression of multiple cardiac diseases, with oxidative stress identified as a central pathophysiological mechanism. Previous animal experiments demonstrated that citronellal (CT), administered at 200 mg/kg in rats, exerted significant cardioprotective effects. However, the molecular mechanisms underlying these effects remain unclear. This study aimed to investigate the role of CT in mitigating myocardial senescence and to elucidate its mechanistic pathways. Doxorubicin-induced myocardial senescence mouse models and H9C2 cardiomyocyte senescence models were established. SA-β-gal staining, Western blotting, immunofluorescence, and immunohistochemistry were employed to evaluate senescence and oxidative stress markers. Network pharmacology analysis and molecular docking were conducted to predict CT targets. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were performed to identify potential mechanisms of action. CT treatment significantly reduced myocardial oxidative stress levels, ameliorated senescent phenotypes in both in vivo and in vitro models, and enhanced mitophagy by activating the AMPKα-mediated PINK1/Parkin pathway. Bioinformatics analyses further supported the involvement of oxidative stress resistance and mitophagy regulation as central mechanisms underlying CT's cardioprotective effects. Citronellal effectively alleviates cardiomyocyte senescence by reducing oxidative stress and promoting mitophagy through activation of the AMPKα–PINK1/Parkin signaling pathway. These findings provide experimental evidence supporting CT as a promising cardioprotective agent and highlight a novel mechanism of action that may inform future therapeutic strategies for cardiac aging and related diseases.
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PubMed
高影响因子
Microbial Biotechnology
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2021
·Vol. 15
·期 5
·1486-1498
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IF 5.813
Baoqi Zhang,Han Du,Yanqiu Zheng,Jiale Sun,Yu Shen,Jinping Lin,Dongzhi Wei
Summary Bioproduction of optical pure ( R )-citronellal from ( E /Z)-citral at high substrate loading remains challenging. Low catalytic efficiency of ( R )-stereoselective ene reductases towards crude citral mixture is one of the major bottlenecks. Herein, a structure-based engineering strategy was adopted to enhance the catalytic efficiency and stereoselectivity of an ene reductase (OYE2p) from Saccharomyces cerevisiae YJM1341 towards ( E / Z )-citral. On basis of homologous modelling, molecular docking analysis and alanine scanning at the binding pocket of OYE2p, a mutant Y84A was obtained with simultaneous increase in catalytic efficiency and stereoselectivity. Furthermore, site-saturation mutagenesis of Y84 yielded seven mutants with improved activity and stereoselectivity in the ( E / Z )-citral reduction. Among them, the variant Y84V exhibited an 18.3% and 71.3% rise in catalytic efficiency ( k cat / K m ) for ( Z )-citral and ( E )-citral respectively. Meanwhile, the stereoselectivity of Y84V was improved from 89.2% to 98.0% in the reduction in ( E / Z )-citral. The docking analysis and molecular dynamics simulation of OYE2p and its variants revealed that the substitution Y84V enabled ( E )-citral and ( Z )-citral to bind with a smaller distance to the key hydrogen donors at a modified ( R )-selective binding mode. The variant Y84V was then co-expressed with glucose dehydrogenase from Bacillus megaterium in E . coli D4, in which competing prim -alcohol dehydrogenase genes were deleted to prevent the undesired reduction in the aldehyde moiety of citral and citronellal. Employing this biocatalyst, 106 g l −1 ( E / Z )-citral was completely converted into ( R )-citronellal with 95.4% ee value and a high space-time yield of 121.6 g l −1 day −1 . The work highlights the synthetic potential of Y84V, which enabled the highest productivity of ( R )-citronellal from ( E / Z )-citral in high enantiopurity so far.
研究领域:
生物催化
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酶工程
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分子生物学
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生物化学
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DOI
PubMed
高影响因子
TOXICON
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2022
·
IF 3.035
Mao Zhang,Hui Li,Addo Keren Agyekumwaa,Yigang Yu,Xinglong Xiao
Staphylococcus aureus ( S. aureus ) is known to be one of the most common foodborne pathogens capable of secreting a wide range of exotoxins such as enterotoxin , which severely threatens the health of consumers. Over the past few years, the development of safe and effective strategies in inhibiting the growth and enterotoxins generation of S. aureus in food turns out to be the research focus and emphasis. This research explores citronellal (CIT), a native compound with extensive existence in spices, which could effectively inhibit the growth and enterotoxins generation of S. aureus (ATCC 29213). Results from minimal inhibitory concentration (MIC), minimum bactericidal concentration (MBC), and time-kill curves, showed that CIT could tremendously inhibit the growth of S. aureus . Analysis on hemolysin showed that CIT at sub-MIC could significantly (p < 0.05) inhibit the hemolytic activity of S. aureus . As revealed by the results of ELISA , the production of enterotoxins in the culture supernatant and pork meat decreased significantly (p < 0.05) after exposure to CIT at sub-MIC. Furthermore, a significant (p < 0.05) decrease in dose-dependent was found in the transcription levels of virulence-related genes. In all, CIT proved to be a possible inhibitor of the growth and enterotoxins production of S. aureus with highly promising application in the food industry.
研究领域:
食品科学
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微生物学
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毒理学
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药理学
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DOI
PubMed
高影响因子
POSTHARVEST BIOLOGY AND TECHNOLOGY
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2026
·
IF 6.8
Xiumei Chen,Chen Zhu,Junyu Chen,Chaobo Bing,Qiuli Ouyang,Xiangrong Zhu,Nengguo Tao
The citrus green mold caused by Penicillium digitatum is a wound-mediated disease considered the main cause of post-harvest citrus fruit spoilage. In this study, an antifungal composite aerogel based on carboxymethyl cellulose (CMC) and polyvinyl alcohol (PVA) was prepared and used to controlling green mold in citrus fruit with a combination of cinnamaldehyde and citronellal (CC). Characterization revealed that the CMC/PVA aerogel had a three-dimensional porous structure, and hydrogen-bond interactions existed between -C O groups of CMC and -OH groups of PVA. The composite aerogels had lower porosity, higher density and lower hygroscopicity performance than raw CMC aerogels. When loaded with CC (4:1, v/v), the aerogel enabled continuous release to fifteen days, with its maximal release rate on the second day. The release profiles of the CMC/PVA-CC were fitted well by the Korsmeyer–Peppas model and the release behavior was governed by Fickian diffusion. In vitro and in vivo tests showed that these aerogels inhibited P. digitatum growth in a concentration-dependent manner. Moreover, CMC/PVA composite aerogels loaded with 0.01 μL L −1 CC suppressed postharvest green mold decay in citrus fruit, demonstrating efficacy comparable to that of the 0.025 % prochloraz, while maintaining fruit quality after storage of 30 d. These results indicated that CC-loaded CMC/PVA composite aerogels can serve as a sustained release and eco-friendly alternative to conventional chemical fungicides for managing postharvest diseases in citrus fruit.
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高影响因子
APPLIED BIOCHEMISTRY AND MICROBIOLOGY
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2025
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IF 1.1
Wu S. J.,Chen F.,Ma X. J.,Sun S. Y.,Yan H. D.,Li T. T.,Qiu L. Q.
( R )-citronellal is a key intermediate in the synthesis of L-menthol, one of the world’s three most important spices. It can be obtained by asymmetric reduction of ( E / Z )-citral using old yellow enzymes (OYEs), but catalysis by the wild-type enzyme suffers from poor enantioselectivity as well as low efficient conversion. A protein-directed evolution technique was used to enhance the activity and enantioselectivity of Ct OYE from Chroococcidiopsis thermalis . Upon the sequence alignment and molecular docking results, two amino acid residues at positions 103 and 351 were selected as saturation mutation sites. Finally, three single substitution variants of Ct OYE-W103E, Ct OYE-W103M, Ct OYE-W103P with improved enantioselectivity ( ee > 99%) and moderate conversion (>71%), and one double substitution variant Ct OYE-Y351Q/W103E with high enantioselectivity ( ee > 99%) and high conversion (83.2%) for reduction of ( E / Z )-citral were obtained.
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高影响因子
Bioresources and Bioprocessing
·
2018
·Vol. 5
·期 1
·1-12
·
IF 0
Zheng Liandan,Lin Jinping,Zhang Baoqi,Kuang Yuyao,Wei Dongzhi
A NAD(P)H-dependent enoate reductase (OYE2p) from Saccharomyces cerevisiae YJM1341 was discovered by genome data mining for asymmetric reduction of (E/Z)-citral to (R)-citronellal with high enantioselectivity.This enzyme was heterologously expressed in E. coli and characterized for its biocatalytic properties. OYE2p was identified with reduction activities toward a diverse range of ɑ,β-unsaturated compounds bearing conjugated aldehyde, ketone, imide, carboxylic acid and ester.OYE2p showed the highest specific activity at 40 °C and a pH optimum at 7.0–8.0. The stability of OYE2p was rather pH-independent, and the half-life time values of the enzyme at pH 6.0–8.0 were more than 257 h. With regard to the reduction of (E)-citral and (Z)-citral, OYE2p exhibited different selectivity patterns. (E)-citral was exclusively reduced to (R)-citronellal by OYE2p in ≥ 99% ee, which was independent on pH. OYE2p produced both enantiomers of citronellal from (Z)-citral, but showed (R)-citronellal formation tendency, and the ee value of (R)-citronellal was affected by pH in the reaction system. Accordingly, the ee values for (R)-citronellal formation increased with the increasing levels of E-isomer in the (E/Z)-citral mixture as well as the increase of pH. Under the reaction conditions (30 °C and pH 8.6), using purified OYE2p as catalyst, 200 mM (E/Z)-citral (an approximately 10:9 mixture of geometric E-isomer and Z-isomer) was efficiently converted to (R)-citronellal with 88.8% ee and 87.2% yield.All these positive features demonstrate high potential of OYE2p for practical synthesis of (R)-citronellal and in asymmetric reduction of activated alkenes.
研究领域:
生物化学
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酶工程
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合成生物学
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INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
·
2024
·
IF 7.7
Ce Shi,Li Jia,Hongxun Tao,Changzhu Li,Tariq Aziz,Majid Alhomrani,Haiying Cui,Lin Lin
Harbin red sausage is a traditional local pork meat product in China, but it is susceptible to microbial contamination and lipid oxidation, leading to quality deterioration. Herein, guar gum (GG)/chitosan (CS) edible films functionalized with citronellal/hydroxypropyl- β -cyclodextrin inclusion complex (CIT/HP β CD-IC) were fabricated for Harbin red sausage preservation. Results demonstrated CIT/HP β CD-IC was successfully prepared and observed by SEM due to the bathochromic shift of maximum absorption peak of CIT, and the formation of new bonds was confirmed by FTIR analysis, suggesting the embedding of CIT into HP β CD cavity. The changes of functional groups stretching vibrations suggested successful loading of CIT/HP β CD-IC into the GG/CS edible films. Furthermore, the incorporation of CIT/HP β CD-IC enhanced the microstructural, mechanical and barrier properties, and improved the antibacterial activities, biodegradability and thermal stability of the GG/CS edible films. Particularly, the GG/CS edible films incorporated with 1 % CIT/HP β CD-IC (GG/CS-IC 1 %) enhanced the storage stability of Harbin red sausage at 4 °C by decreasing the weight loss rate, maintaining the pH, color, and textural stabilities, retarding the microbial growth and lipid oxidation of the sausage samples. Findings here suggested that GG/CS-IC 1 % edible films showed great potential as novel multi-functional edible packaging materials for Harbin red sausage preservation.
研究领域:
食品科学
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材料科学
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生物技术
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DOI
PubMed
高影响因子
Food Frontiers
·
2023
·
IF 9.9
Chenlinrui Yue,Fengjiao Fan,Yinji Chen,Yong Fang,Hongkai Xie
Biodegradable active packaging films with antimicrobial and antioxidant functions have received much attention due to the food safety and plastic pollution issues. This study prepared and characterized a novel bioactive binary composite film based on gelatin (Ge) and dextran (Dx) incorporated with various concentrations (0%, 0.3%, 0.6%, 0.9%, 1.2%, w/w) of citronellal (Cit) and α-tocopherol (Toc). The thickness, L *, b *, total color difference, water contact angle, opacity, and crystallinity of film significantly increased, whereas the film solubility, swelling degree, water vapor permeability, tensile strength, elongation at break, and a * of film significantly decreased with the increase of Cit and Toc concentrations. However, the addition of Cit and Toc had no significant effect on the film's moisture content and thermal stability. Fourier transforms infrared spectral pattern indicated that physical interactions (H-bonding) between biopolymer matrix and active fillers were formed. Scanning electron microscope images showed that the film added with active fillers had a rough surface and a spongelike cross-section structure compared to control film. Moreover, the release curves of Cit and Toc showed that the film in 95% ethanol solution had the longest release time. Furthermore, the film added with Cit and Toc effectively inhibited the growth of pathogenic and spoilage bacteria ( Shewanella putrefaciens and Escherichia coli ) and quenched the free radicals (2,2-diphenyl-1-picrylhydrazyl and 2,2′-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid)). Overall, it can be inferred that the Ge–Dx-based composite film incorporated with Cit and Toc is a potential antibacterial and antioxidant biodegradable material for food packaging applications to maintain food quality.
研究领域:
材料科学
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食品科学
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生物技术
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DOI
高影响因子
危险品化学品经营许可证（带存储）