APPLIED AND ENVIRONMENTAL MICROBIOLOGY
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2024
·Vol. 91
·期 1
·
IF 3.9
Ji Zeng,Hao Wang,Yuxi Xu,Jianying Han,Yannan Li,Shu'an Wen,Changbu Wu,Dani Li,Zheng Liu,Xiaokang Zhang,Guo-Bao Tian,Min Dong
Clostridioides difficile is an obligate anaerobic, Gram-positive bacterium that produces toxins. Despite technological progress, conducting gene expression analysis of C. difficile under different conditions continues to be labor-intensive. Therefore, there is a demand for simplified tools to investigate the transcriptional and translational regulation of C. difficile. The cell-free gene expression (CFE) system has demonstrated utility in various applications, including prototyping, protein production, and in vitro screening. In this study, we developed a C. difficile CFE system capable of in vitro transcription and translation (TX-TL) in the presence of oxygen. Through optimization of cell extract preparation and reaction systems, we increased the protein yield significantly. Furthermore, our observations indicated that this system exhibited higher protein yield using linear DNA templates than circular plasmids for in vitro expression. The prototyping capability of the C. difficile CFE system was assessed using a series of synthetic Clostridium promoters, demonstrating a good correlation between in vivo and in vitro expression. Additionally, we tested the expression of tcdB and tcdR from clinically relevant C. difficile strains using the CFE system, confirming higher toxin expression of the hypervirulent strain R20291. We believe that the CFE system can not only serve as a platform for in vitro protein synthesis and genetic part prototyping but also has the potential to be a simplified model for studying metabolic regulations in Clostridioides difficile.
研究领域:
微生物学
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分子生物学
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生物技术
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DOI
高影响因子
CARBOHYDRATE POLYMERS
·
2014
·
IF 3.916
Jingfen Han,Jia Cai,Wuyinga Borjihan,Tsogzolmaa Ganbold,Tariq M. Rana,Huricha Baigude
RNA interference (RNAi) down-regulates gene expression post-transcriptionally, which is a therapeutically significant phenomenon that could potentially reduce the level of disease related proteins that are undruggable by conventional small molecular approaches. However, clinical application of small interference RNAs (siRNAs) requires design of potent siRNA sequences and development of safe and efficient delivery systems. To create a biocompatible siRNA delivery agent, we chemically modified natural polysaccharide curdlan in a regioselective manner to introduce amino group in the glucose units. The resulting 6-amino-curdlan (6AC) is water soluble and forms nanoparticles upon complexing with siRNAs. The novel curdlan-based nanoparticles efficiently delivered siRNAs to human cancer cells and mouse primary cells, and reduced 70–90% of target mRNA level. Moreover, 6AC nanoparticles delivered siRNA targeting eGFP to mouse embryonic stem (mES) cells stably expressing eGFP, and produced substantial reductions of GFP protein level. The novel curdlan-based nanoparticle is a promising vehicle for delivery of short RNAs to knock down endogenous mRNAs.
研究领域:
生物技术
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分子生物学
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药物递送
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DOI
PubMed
高影响因子
Animal
·
2017
·Vol. 12
·期 3
·491-500
·
IF 1.921
Q. Liu,C. Wang,G. Guo,W. J. Huo,Y. L. Zhang,C. X. Pei,S. L. Zhang,H. Wang
Isovalerate supplements could stimulate rumen development by improving morphology and function of rumen mucosa, and then promote the growth of calves. This study was done to evaluate the effects of isovalerate supplements on morphology and functional gene expression of rumen mucosa in dairy calves. In total, 48 Chinese Holstein male calves with 15 days of age and 45.1±0.36 kg of BW were randomly assigned to four groups. The treatments were: control, low-isovalerate, moderate-isovalerate and high-isovalerate with 0, 3, 6 and 9 g isovalerate per calf per day, respectively. Supplementary isovalerate was hand-mixed into milk in pre-weaning calves and into concentrate portion in post-weaning calves. The study consisted of a 15-day-adaptation period and a 60-day-sampling period. Calves were weaned at 60 days of age. Three calves were slaughtered from each of the four treatments at 30, 60 and 90 days of age. The weight of body and stomach were measured, samples of ruminal tissues and blood were analyzed. Total stomach weight, total stomach to BW ratio, rumen wall and keratinized layer thickness, serum growth hormone and IGF-1 for both pre- and post-weaning calves increased linearly with increasing isovalerate supplements. Rumen to total stomach weight ratio, the length and width of rumen papillae, and serum β-hydroxybutyrate increased linearly for post-weaning calves. However, abomasum weight to total stomach weight ratio decreased linearly for both pre- and post-weaning calves. The relative messenger RNA expression for growth hormone receptor, IGF-1 receptor and 3-hydroxy-3-methylglutaryl-CoA synthase 1 in rumen mucosa increased linearly for post-weaning calves. Our results suggested that isovalerate supplements promoted rumen development in a dose-dependent manner. The optimum dose was 6.0 g isovalerate per calf per day.
研究领域:
动物科学
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分子生物学
✓ 已验证
DOI
PubMed
高影响因子
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
·
2019
·
IF 3.571
Hengqian Lu,Haiqin Chen,Xin Tang,Qin Yang,Hao Zhang,Yong Q. Chen,Wei Chen
The objective of the present study was to reveal the effects of four types of nitrogen sources (soymeal, yeast extract, KNO3, and ammonium tartrate) on the lipid metabolism of the oleaginous fungus Mortierella alpina using untargeted lipidomics, targeted fatty acid, and reverse transcription quantitative polymerase chain reaction (RT-qPCR) analysis. Our results showed clear differences in the contents and compositions of lipids between four types of nitrogen sources. Soymeal and ammonium tartrate supplementation favored the accumulation of triglycerides with arachidonic acid (ARA) and C16–18 fatty acids, respectively. These results were further validated by our targeted fatty acid analysis. RT-qPCR analysis of related genes in M. alpina between the four nitrogen source conditions found that soymeal supplementation dramatically increased the expression of GPAT, ELOVL, and Δ12/Δ6 desaturase. Our findings provided new insights into the regulation of lipid biosynthesis in M. alpina and potential avenues for genetic manipulation and highlighted the importance of an optimal nitrogen source for ARA-rich oil production.
研究领域:
生物化学
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微生物学
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分子生物学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
SOUTH AFRICAN JOURNAL OF BOTANY
·
2024
·
IF 2.7
Xiaoyu Ren,Haiou Wang,Wenxuan Wang,Wenxiao Zhang,Qian Li
Combined contamination of Organophosphate esters (OPEs) and heavy metal has been concerning on their potential threaten to human health through food chain. In order to estimate effects of OPEs and heavy metal's combined contamination on crop, the growth of rice seedlings, pollutant accumulations and their involved in transcript regulation pathway under OPEs and Cd contamination were investigated. In results, only single OPEs pollution improved the biomass of rice, while single Cd pollution inhibited the growth of plants. Although the combined contaminations of OPEs and Cd also showed negative effect for plant growth, OPEs could alleviate the negative effect of Cd on rice. The Octanol-water partition coefficients (log K ow ) of OPEs were positively correlated with root concentration factor (RCF) values in the trend line and negatively correlated with shoot concentration factor (SCRF) and translocation factor (TF). OPEs and Cd inferred with each other's accumulations in plants through reducing RCF, SCRF and TF. After addition of Cd, the expressions of OsTIL, OsLTP4, OsLTPL1 , and OsMLP423 genes involved in OPEs accumulation were downregulated. Moreover, the presence of Cd also affected the binding sites and binding energy of the four proteins with OPEs. Meanwhile, the expression of OsNramp1, OsNramp5, OsHMA2, OsGS1 , and OsPCS1 genes involved in Cd accumulation and toleration were also downregulated after addition of OPEs. These results suggested that OPEs and Cd directly repressed each other gene transcript involved in accumulation and toleration process.
研究领域:
环境科学
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植物学
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分子生物学
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产品关联
DOI
高影响因子
Biomolecules
·
2025
·Vol. 15
·期 4
·492
·
IF 4.8
Xia Xu,Chunguang Cui,Xin Du,Jine Chen,Xiuling He,Linbao Zhu,Chengjie Hu,Fang Xu,Chenkai Ma,Shaofang Yu,Xingjian He,Houhui Song,Yongqiang Wang
Lipid metabolism serves as the primary energy source for organisms. Silkworm eggs for spring use are divided into two types: autumn-produced eggs for next spring rearing (AS) and spring-produced eggs for next spring rearing (SS). Production practice revealed significant differences in hatching rates between these two types of silkworm production strain QiufengA. In this study, we identified differentially expressed genes (DEGs) primarily enriched in energy metabolism pathways. In particular, the PPARs are involved in energy regulation through lipid metabolism. Furthermore, both AS and SS contained the same eight long-chain fatty acids but in different amounts. Interference with PPARs activity in silkworm eggs disrupted the expression of key genes in this pathway, resulting in a significant decrease in hatching rate. Additionally, knockdown of the pathway key geneBmPlin4led to the reduction in lipid droplets. In conclusion, PPARs regulates the hatching rate of silkworms mainly by affecting lipid metabolism. This study proved the importance of PPARs for hatching and identifies them as potential target genes for population control.
研究领域:
昆虫学
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分子生物学
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遗传学
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产品关联
DOI
PubMed
高影响因子
Food Bioscience
·
2025
·
IF 5.9
Lei Chen,Zhuomin Zhang,Zihan Guan,Jing Sun,Yanhui Sun,Mengmeng Xu,Ka-Hing Wong,Guiyang Shi,Zhongyang Ding
Ergothioneine (EGT) production in Pleurotus tuber-regium was significantly enhanced through Na 2 SeO 3 -induced oxidative stress, in combination with optimized carbon sources and precursor amino acids. Initially, glucose and rhamnose (20 g/L each) as dual carbon sources increased EGT synthesis. Supplementation with 0.75 mM cysteine and 1 mM methionine further promoted EGT biosynthesis by providing essential precursors. Additionally, the introduction of 0.3 mM Na 2 SeO 3 induced oxidative stress, leading to an EGT accumulation of 4.68 mg/g dry weight (DW) in the mycelium and a yield of 35.27 mg/L, representing a 4.25-fold and 162% increases compared to the control, respectively. RNA-Seq analysis revealed that selenium exposure triggered the upregulation of genes involved in antioxidant defense, reinforcing redox homeostasis and driving EGT accumulation. This study highlights the mechanistic role of selenium in modulating antioxidant pathways, offering a promising strategy for enhancing EGT production in the nutraceutical and biotechnological fields.
研究领域:
生物技术
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营养学
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分子生物学
✓ 已验证
产品关联
DOI
高影响因子
FOOD RESEARCH INTERNATIONAL
·
2023
·
IF 8.1
Danliangmin Song,Ai Jia,Xuehe Qi,Kai Dong,Shiyu Liu,Chaoxin Man,Xinyan Yang,Yujun Jiang
Bacterial biofilm is a protective matrix composed of metabolites secreted by bacteria that envelop bacteria. By forming a biofilm, bacteria can considerably improve their environmental tolerance. In food-related processing environment, different types of microorganisms are often present in biofilms. The main contaminating strain in the powdered infant formula (PIF) processing environment, Cronobacter sakazakii and Staphylococcus aureus continues to pollute the PIF processing environment after biofilm production. This study selected Cronobacter sakazakii with a weak biofilm-forming ability as one of the test organisms. The coexistence of Cronobacter sakazakii and Staphylococcus aureus on the surface of production equipment was simulated to analyze the interaction. Biofilm formation in the co-culture group was significantly higher than the others. In-depth study of the effect of Staphylococcus aureus on the biofilm formation genes of Cronobacter sakazakii . Results show two bacteria can coexist on the surface of a metal device, forming a more compact hybrid biofilm structure. Under co-culture conditions, S. aureus increased bcsA and fliD expression in Cronobacter sakazakii , whereas decreased bcsC expression. Signaling molecules produced by Staphylococcus aureus (Autoinducer 2) significantly promoted the biofilm formation of Cronobacter sakazakii at the concentration of 0–500 ng/mL (0.099–0.177) and up-regulated the expression of bcsA , filD and flhD genes.
研究领域:
微生物学
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食品科学
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产品关联
DOI
PubMed
高影响因子
PESTICIDE BIOCHEMISTRY AND PHYSIOLOGY
·
2023
·
IF 4.7
Shaoquan Liu,Jianlu Zhang,Ruijiao Li,Chunyun Zhang,Lixin Wang,Hao Liang,Guangpeng Feng,Dongmei Xiong
Due to chemical and photochemical stability, triazophos has been frequently detected in rivers and oceans over the years with extensive use for pest control in agriculture , and it has become a worldwide ecological concern to the aquatic environment . Until now, fewer data are available regarding the potential long-term adverse effects of triazophos on aquatic invertebrates , which plays an essential role in aquatic food webs , as a key group for water ecosystems. In this experiment, the F 1 - and F 2 progenies of Daphnia magna were recovered when daphnias (F 0 ) exposure to triazophos at environmental-related concentrations (0.1 and 1.0 μg/L) for 21 d; and the indexes related to phenotypic traits, reproduction and gene expression were measured in tested animals. The results showed that heart rate and total number of neonates in exposed F 0 -daphnias were significantly lower than those of control group, and the detoxification genes ( HR96 and P-gp ) were up-regulated while genes related reproduction ( Vtg ) and molting ( Nvd and Shd ) were significantly down-regulated. The heart rate and individual size of F 1 -daphnias (<24 h) were significantly reduced in the treatment group. After 21-d recovery, the heart rate and expression of HR96 , P-gp, Vtg, Nvd and Shd were declined in F 1 -daphnias. There was no obvious difference of morphological traits and heart rate between treatment and control in F 2 -daphnias (<24 h). In summary, daphnias (F 0 ) exposure to triazophos with environmental dose could raise toxic effects on its offspring (F 1 ), which is mainly manifested by reduced heart rate, the accumulated number and individual size of offspring and decreased expression of genes related to molting and reproduction.
研究领域:
环境毒理学
·
水生生态学
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产品关联
DOI
PubMed
高影响因子
COMPARATIVE BIOCHEMISTRY AND PHYSIOLOGY C-TOXICOLOGY & PHARMACOLOGY
·
2024
·
IF 3.9
Fang Chanlin,Di Shanshan,Wang Caihong,Hu Qinglian,Jin Yuanxiang
The increasing release of tire-derived particles, particularly those containing N-(1,3-dimethylbutyl)- N ′-phenyl- p -phenylenediamine (6PPD), into the environment has raised concerns regarding their ecological impact. This study aims to elucidate the toxicological effects of 6PPD on the metabolism in early developmental stage of zebrafish. Larval zebrafish were exposed to 10 and 100 μg/L 6PPD, and some endpoints in biochemical parameters, gene expression, and metabolism were analyzed. The results showed that 6PPD exposure disrupted glucolipid metabolism in zebrafish larvae, evidenced by increased triglyceride (TG) levels and decreased glucose content. Nile red staining indicated significant lipid accumulation in the liver and intestines. Additionally, RT-qPCR analysis revealed the upregulation of genes involved in lipid synthesis and metabolism, such as ppar-γ and fas , and downregulation of glycolysis-related genes like pk and gk . Furthermore, the untargeted metabolomics technique was used to identify a total of 220 differentially expressed metabolites (DEMs) with changes in amino acid metabolism, lipid metabolism, and the TCA cycle. KEGG pathway enrichment analysis highlighted disruptions mainly in Taurine and hypotaurine metabolism, Arginine and proline metabolism, and Histidine metabolism, which played very important roles on energy metabolism in zebrafish. The results provided some critical insights into the ecological risks associated with 6PPD.
研究领域:
环境科学
·
毒理学
·
生物化学
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产品关联
DOI
PubMed
高影响因子
JOURNAL OF HAZARDOUS MATERIALS
·
2025
·
IF 11.3
Minmin Zhang,Chenyan Liao,Zhonghuai Zhang,Lichao Lu,Jianmeng Chen,Dongzhi Chen
Chlorobenzene (CB) biodegradation is challenging due to its hydrophobic characteristics. Addition of silicone oil and surfactants are commonly used methods to enhance biodegradation efficiency by improve CB gas-liquid mass transfer efficiency. However, both approaches introduced new chemicals into the reactor, either as carbon sources for microorganisms or requiring regeneration. This study established a synergistic reactor by applying micro-nano bubbles (MNBs) with biodegradation process in a stirred tank bioreactor (STB) to purify CB waste gas. Experimental results demonstrate a reduction in start-up time by 2 days and a 13.5 % increase in overall degradation rate in synergistic reactor. The mass transfer fraction of CB ( β * s ) was increased by 13.33 %. Additionally, the synergistic reactor showed improved system stability and microbial activity, evidenced by the increased Zeta potential, extracellular polymer substances (EPS) secretion, and protein content. Notably, MNBs upregulate genes involved in aromatic ring hydroxylation and dehalogenation processes and promoted the enzyme SCACT (EC2.8.3.18) within the genus Acidovorax , which enhanced intracellular coenzyme A activity and facilitated chlorobenzene degradation in this genus, thereby enhancing CB degradation efficiency. These results indicate that MNBs can significantly improve the biodegradation performance of CB waste gas, offering a promising strategy for industrial applications.
研究领域:
环境科学
·
微生物学
✓ 已验证
DOI
PubMed
高影响因子
FOOD RESEARCH INTERNATIONAL
·
2024
·
IF 7
Luyao Fan,Hongchao Dai,Wenyuan Zhou,Lei Yuan,Jia Yang,Zhenquan Yang,Xin-an Jiao
Bacillus licheniformis , a quick and strong biofilm former, is served as a persistent microbial contamination in the dairy industry. Its biofilm formation process is usually regulated by environmental factors including the divalent cation Ca 2+ . This work aims to investigate how different concentrations of Ca 2+ change biofilm-related phenotypes (bacterial motility, biofilm-forming capacity, biofilm structures, and EPS production) of dairy B . licheniformis strains. The Ca 2+ ions dependent regulation mechanism for B . licheniformis biofilm formation was further investigated by RNA-sequencing analysis. Results revealed that supplementation of Ca 2+ increased B . licheniformis biofilm formation in a dose-dependent way, and enhanced average coverage and thickness of biofilms with complex structures were observed by confocal laser scanning microscopy. Bacterial mobility of B. licheniformis was increased by the supplementation of Ca 2+ except the swarming ability at 20 mM of Ca 2+ . The addition of Ca 2+ decreased the contents of polysaccharides but promoted proteins production in EPS, and the ratio of proteins/polysaccharides content was significantly enhanced with increasing Ca 2+ concentrations. RNA-sequencing results clearly indicated the variation in regulating biofilm formation under different Ca 2+ concentrations, as 939 (671 upregulated and 268 downregulated) and 951 genes (581 upregulated and 370 downregulated) in B. licheniformis BL2-11 were induced by 10 and 20 mM of Ca 2+ , respectively. Differential genes were annotated in various KEGG pathways, including flagellar assembly, two-component system, quorum sensing, ABC transporters, and related carbohydrate and amino acid metabolism pathways. Collectively, the results unravel the significance of Ca 2+ as a biofilm-promoting signal for B. licheniformis in the dairy industry.
研究领域:
微生物学
·
食品科学
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产品关联
DOI
PubMed
高影响因子
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