Synthetic and Systems Biotechnology
·
2025
·
IF 4.4
Ruizhou Tang,Jiahuan Li,Xiaole Yang,Xia Tian,Zehui Wang,Xuning Zhang,Tingting Li
Aminotransferases are promising green biocatalysts for the synthesis of chiral amines, yet their limited catalytic efficiencies restrict broader industrial applications. In this study, a novel ( R )-amine transaminase, FalAT, was identified from Fusarium albosuccineum through genome mining. FalAT exhibited optimal activity at 30 °C and pH 7.0 and catalyzed the conversion of 1-Boc-3-piperidone to ( R )-1-Boc-3-aminopiperidine with >99 % enantiomeric excess. To efficiently enhance its catalytic performance, a Substrate–Protein Interaction Network (SPIN) strategy was implemented, integrating structure-guided analysis, molecular docking, virtual saturation mutagenesis, and dual energy–distance filtering. SPIN first screened and constructed a mutational library covering 70 amino acid positions, which was subsequently narrowed to 9 key residues, ultimately yielding 15 candidate mutants for experimental validation. Experimental results showed that five mutants exhibited higher catalytic activity than the wild-type enzyme, among which R126A was the most effective, displaying approximately a 4-fold increase in catalytic activity and a 13-fold enhancement in catalytic efficiency ( k ca t /K m = 2.05 s −1 mM −1 ). Molecular dynamics simulations revealed that the R126A mutation expanded the active-site cavity, alleviated steric hindrance, and strengthened hydrophobic interactions, thereby improving substrate binding and catalytic turnover. Furthermore, substrate profiling demonstrated that FalAT possesses moderate substrate promiscuity. Overall, the SPIN strategy significantly improved the catalytic performance of FalAT while markedly reducing experimental workload, providing an efficient and generalizable approach for the directed evolution of ( R )-amine transaminases for the green synthesis of chiral amines.
研究领域:
酶工程
·
合成生物学
·
生物催化
·
结构生物学
·
分子设计
·
绿色化学
·
药物代谢
✓ 已验证
产品关联
DOI
PubMed
高影响因子
JOURNAL OF THE SCIENCE OF FOOD AND AGRICULTURE
·
2016
·Vol. 97
·期 4
·1342-1348
·
IF 2.076
Cuiqiong Wang,Zhongyuan Li,Hui Wang,Haiyan Qiu,Minghui Zhang,Shuang Li,Xuegang Luo,Yajian Song,Hao Zhou,Wenjian Ma,Tongcun Zhang
BACKGROUND Contamination of food and feed by aflatoxin B1 (AFB1) poses serious economic and health problems worldwide, so the development of biological methods for effective AFB1 degradation is strongly required. RESULTS Among three AFB1-degrading microorganisms isolated from moldy peanut, Fusarium sp. WCQ3361 could remove AFB1 extremely effectively, with a degradation ratio of 70.20% after 1 min and 95.38% after 24 h. Its degradation ratio was not much affected by temperature change (0–90 °C) and it also displayed excellent thermostability, maintaining 99.40% residual activity after boiling for 10 min. Since protease K could reduce the AFB1 degradation ratio by 55.15%, it is proposed that the effective component for AFB1 degradation is a protein. The AFB1 degradation ability of Fusarium sp. WCQ3361 was further verified by feed stock detoxification and the MTT test with HepG2 cells. In addition, no degradation products were detected by preliminary liquid chromatography/mass spectrometry, suggesting that AFB1 might be metabolized to products with different chemical characteristics from AFB1. CONCLUSION Fusarium sp. WCQ3361 is the first reported AFB1 degradation fungus belonging to the genus Fusarium with broad working temperature range, excellent thermostability and high activity, which provides a potential highly useful solution for dealing with AFB1 contamination in the human diet and animal feed. © 2016 Society of Chemical Industry
研究领域:
食品科学
·
微生物学
·
毒理学
✓ 已验证
DOI
PubMed
高影响因子
TRENDS IN BIOTECHNOLOGY
·
2026
·
IF 16.6
Ya-Wen Li,Xin-Yu Chen,Hui-Zhi Lu,Ji-Bao Yao,Zhi-Kui Nie,Tian-Qiong Shi
Gibberellic acid (GA3) is primarily produced by the liquid fermentation of Fusarium fujikuroi , yet its complex metabolic network limits rational strain engineering. Here, we strengthened the nonhomologous end joining (NHEJ) pathway and mapped the distribution of integration to establish a genome-scale insertional mutagenesis library in F. fujikuroi . The library enabled rapid identification of ten novel genomic loci that support stable heterologous gene expression, thereby expanding the available genetic toolkit. We further applied the library to uncover functional determinants of GA3 biosynthesis and acetic acid tolerance, identifying previously uncharacterized gene targets. Reverse engineering of these targets yielded a GA3-overproducing strain capable of tolerating and utilizing acetic acid wastewater. In a 5-l bioreactor, the engineered strain NH4 reached a GA3 titer of 3.32 g/l, 3.05-fold higher than the control strain. This study establishes an efficient NHEJ-mediated genome-scale mutagenesis platform for F. fujikuroi and provides more genetic targets for constructing high-yield GA3 cell factories.
✓ 已验证
产品关联
DOI
PubMed
高影响因子
FOOD CHEMISTRY
·
2026
·
IF 10.4
Jingyi Tang,Haibin Liao,Xueqin Fu,Yonghong Xie,Zhengrong Zou,Zaizhi Liu
Methyleugenol was encapsulated via ultrasonic assisted microencapsulation method. • High yield and encapsulation efficiency were obtained with the proposed method. • Methyleugenol microcapsules had good thermal stability and sustained-release ability. • Methyleugenol microcapsules showed remarkable antifungal activity on F. avenaceum . Methyleugenol microcapsules were successfully fabricated using soybean protein isolate and sodium alginate as wall materials through ultrasonic assisted microencapsulation method. A real yield of 95.45 ± 2.55% and an encapsulation efficiency of 76.95 ± 3.03% were achieved under the optimum conditions derived from Plackett–Burman and Box–Behnken designs. Physicochemical characterization confirmed effective methyleugenol encapsulation, with the microcapsules exhibiting irregular morphology, uniform particle size distribution, and distinctive wrinkled surface textures. Thermogravimetric curve indicated that the thermal stability of methyleugenol was obviously enhanced by the microencapsulation process. After microencapsulation, the antifungal activity of methyleugenol was greatly improved, with its microcapsules exhibiting comparable efficacy to nystatin against Fusarium avenaceum , demonstrating the minimum inhibitory concentration (MIC) of 0.10 mg/mL and 0.07 mg/mL, respectively. These findings provide a scientific basis for exploring the potential applications of ME microcapsules in food preservation and related industrial sectors.
✓ 已验证
产品关联
DOI
高影响因子
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
·
2025
·
IF 8.5
Xiao-hua Chen,Xiao-min Li,Jie Deng,Jin-mei Li,Di-qiu Liu
Fusarium wilt is a significant problem restricting the growth of the lily industry. However, Lilium regale Wilson is a valuable germplasm resource because of its Fusarium Wilt resistance. In this study, six endo -β-1,4-glucanase genes ( Cel1-Cel6 ) in L. regale were identified among the differentially expressed genes reveled by omics data for L. regale infected Fusarium osysporum , which causes Fusarium wilt. Interestingly, the abundance of a few proteins and the expression of endo -β-1,4-glucanase genes, including LrCel1 , decreased in L. regale roots infected with F. oxysporum. Thus, the function of LrCel1 in the incompatible interaction between L. regale and F. oxysporum was analyzed. LrCel1 expression was inhibited by F. oxysporum and salicylic acid (SA). Additionally, the subcellular localization of LrCel1 revealed it was a cell membrane protein. The ectopic expression of LrCel1 increased of susceptibility of tobacco to F. oxysporum because of the associated increase in cellulase activity, which, hindered the production of cellulose, callose, and lignin. Moreover, LrCel1 overexpression inhibited the accumulation of SA and jasmonic acid (JA), resulting in the down-regulated expression of genes associated with SA/JA signaling pathways. Conversely, introducing an RNAi vector targeting LrCel1 into L. regale enhanced the resistance to F. oxysporum , likely because of an increase in the synthesis of cellulose, callose, and lignin as well as a decrease in the expression of JA/SA signaling pathway genes. In addition, LrCel1 promoter activity was suppressed by F. oxysporum and Fusarium solani as well as SA. Furthermore, a positive regulator of Fusarium wilt resistance, LrWRKY11, negatively regulated LrCel1 promoter activity. The study findings have elucidated the intricate regulatory network of WRKY transcription factor and SA/JA signaling pathways to inhibit the cellulase activity of endo -β-1,4-glucanase in L. regale , thereby conferring resistance to Fusarium wilt.
研究领域:
植物病理学
·
分子生物学
·
植物遗传学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
BIOSENSORS & BIOELECTRONICS
·
2024
·
IF 10.7
Yehang Wu,Pan Li,Tao Xie,Rui Yang,Rui Zhu,Yulong Liu,Shengyu Zhang,Shizhuang Weng
Determination of Fusarium spores is essential for precision control of Fusarium head blight and ensuring agri-food safety. As a highly sensitive real-time detection technique with rich fingerprint information and little influence from water, surface-enhanced Raman spectroscopy (SERS) has been widely applied in the determination of microorganisms. However, fungi determination faces significant challenges including low sensitivity, and poor specificity. The enhanced quasi-meshing hotspot effect (EQMHE) integrated with the embedded attention residual network (EARNet) was proposed to realize a label-free and accurate SERS determination of various Fusarium spores. The EQMHE can promote the binding of spores and nanoparticles to form numerous hotspots, significantly enhancing signal quality and improving the detection limit by at least four orders of magnitude. The key factors inducing EQMHE were validated through various characterization techniques. Moreover, due to its excellent feature extraction and recognition capabilities, EARNet successfully overcomes the limitations of spectral similarity, achieving determination accuracies of 100% in the training set, 98.33% in the validation set, and 100% in the prediction set for three Fusarium species from actual samples. EARNet requires only a small amount of training data and provides rapid and accurate diagnostics. Throughout the process, the spores do not require culturing or lysing, providing an effective determination method for the practical determination of mixed fungal spores. Overall, the proposed strategy effectively addresses challenges such as the need for fungal spore cultivation, difficulties in SERS hotspot formation, and suboptimal signal quality, and it holds significant promise for applications in disease control, food safety, and agricultural production.
研究领域:
植物病理学
·
食品安全
·
农业科学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Analytical Methods
·
2014
·Vol. 6
·期 17
·6651-6657
·
IF 1.938
Zhiyong Zhao,Suquan Song,Na Liu,Qinxiong Rao,Jiafa Hou,Sarah De Saeger,Aibo Wu
Food safety monitoring and toxicological research of mycotoxins are still in need of large quantities of high-purity deoxynivalenol (DON). To attain this purpose, a rapid, economical and reproducible purification method was developed for large-scale production of DON from rice culture inoculated with a DON-producing Fusarium graminearum strain JYH. The inoculated rice culture was first extracted with acetonitrile–water (84/16, v/v). The extracts were evaporated to dryness on a rotary evaporator after ethyl acetate partitioning and then dissolved in water followed by the final purification procedure through preparative high performance liquid chromatography and montmorillonite treatment. A combined approach of ultraviolet spectrometry (UV), ultra high performance liquid chromatography (UHPLC), mass spectrometry (MS) and nuclear magnetic resonance (NMR) spectroscopy was applied for the multi-dimensional characterization of the target compound. As a result, the recovery of DON from the crude extract to the final product was up to 70%. An amount of 150 mg DON with the desirable purity of 98.93% could be obtained from 100 g of rice culture, which possessed identical immunochemical characteristics compared to a certified commercial DON standard. This proposed strategy might act as a valuable reference to obtain rather expensive compounds in a straightforward way.
研究领域:
食品安全
·
真菌学
·
毒理学
✓ 已验证
产品关联
DOI
高影响因子
INDUSTRIAL CROPS AND PRODUCTS
·
2023
·
IF 5.9
Dicheng Ma,Tao Zhang,Guoxian Wang,Chong Cao,Wei Mu,Beixing Li,Daolong Dou,Feng Liu
Trans -2-hexenal (T2H) shows satisfactory antagonistic activity against Fusarium graminearum , but its strong volatility and poor water solubility limit its further application in Fusarium head blight (FHB) control. In this study, T2H-loaded polyurea microcapsules (PU MCs) with different particle sizes were successfully prepared by an interfacial polymerization method, and the effect of MC encapsulation on the physical property limitations and bioactivity of T2H was evaluated. After being encapsulated, lipophilic T2H could be stably dispersed in water. MC encapsulation reduced the fumigation toxicity of T2H but increased its contact toxicity. The smaller the particle size was, the higher the toxicity of PU MCs to F. graminearum was. After 6 d of application, the inhibition rate of T2H against F. graminearum decreased by 77.26%, while that of T2H-loaded PU MCs with small particle sizes (T2H-loaded PU MCs-S) decreased by 54.29%, indicating that MC encapsulation prolonged the inhibition time of T2H. The results of the inoculation assay showed that MC encapsulation improved the inhibitory effect of T2H on F. graminearum infection and DON production in stored wheat kernels. Furthermore, seed soaking with T2H-loaded PU MCs-S reduced the pathogenic fungal load of wheat seeds and had no significant effect on seed germination. In conclusion, T2H encapsulated in MCs can be developed as a plant protection product for postharvest and presowing treatment of wheat.
研究领域:
植物保护
·
真菌学
·
农药科学
✓ 已验证
产品关联
DOI
高影响因子
Journal of Fungi
·
2026
·Vol. 12
·期 8
·576
·
IF 4.3
Dan-Na Ma,Yihan Niu,Mengli Chen,Jun-Nan Yao,Huiming Wu,Taiying Li
Fusarium wilt, caused by the soil-borne fungusFusarium oxysporumf. sp.lycopersici(Fol), is a devastating disease that severely constrains tomato production worldwide. In this study, an endophytic bacterium, designated ZN-S18, was isolated from healthy tomato stems and identified asBacillus amyloliquefaciensbased on phylogenetic analyses of the 16S rRNA andrpoBgenes. Dual culture assays demonstrated that ZN-S18 exhibited strong antagonistic activity against Fol and significantly inhibited fungal mycelial growth. The cell-free culture filtrate of ZN-S18 markedly suppressed Fol spore germination and mycelial growth, and the inhibitory effect on spore germination was irreversible. LC-MS analyses confirmed that ZN-S18 produces the cyclic lipopeptides surfactin and iturin A. In planta assays further demonstrated that pretreatment with ZN-S18, its culture filtrate, or the combination of surfactin and iturin A significantly reduced Fusarium wilt in tomato seedlings. Moreover, the culture filtrate disrupted fungal cell wall integrity, as indicated by the increased sensitivity of Fol to Congo red stress. Notably, the combined application of surfactin and iturin A exhibited significantly stronger antifungal activity than either compound alone under Congo red-containing conditions, suggesting a synergistic effect on fungal cell wall integrity. Collectively, these findings indicate that the endophytic strainB. amyloliquefaciensZN-S18 is a promising biocontrol agent against tomato Fusarium wilt, primarily through the synergistic action of surfactin and iturin A. This study provides an environmentally friendly alternative to chemical fungicides and establishes a foundation for future investigations into the molecular mechanisms underlying lipopeptide-mediated antagonism.
✓ 已验证
DOI
高影响因子
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
·
2024
·Vol. 25
·期 8
·4514
·
IF 4.9
Chaopu Ding,Yunfei Zhang,Chongbin Chen,Junfang Wang,Mingda Qin,Yu Gu,Shujing Zhang,Lanying Wang,Yanping Luo
In agriculture, soil-borne fungal pathogens, especiallyFusarium oxysporumstrains, are posing a serious threat to efforts to achieve global food security. In the search for safer agrochemicals, silica nanoparticles (SiO2NPs) have recently been proposed as a new tool to alleviate pathogen damage including Fusarium wilt. Hollow mesoporous silica nanoparticles (HMSNs), a unique class of SiO2NPs, have been widely accepted as desirable carriers for pesticides. However, their roles in enhancing disease resistance in plants and the specific mechanism remain unknown. In this study, three sizes of HMSNs (19, 96, and 406 nm as HMSNs-19, HMSNs-96, and HMSNs-406, respectively) were synthesized and characterized to determine their effects on seed germination, seedling growth, andFusarium oxysporumf. sp.phaseoli(FOP) suppression. The three HMSNs exhibited no side effects on cowpea seed germination and seedling growth at concentrations ranging from 100 to 1500 mg/L. The inhibitory effects of the three HMSNs on FOP mycelial growth were very weak, showing inhibition ratios of less than 20% even at 2000 mg/L. Foliar application of HMSNs, however, was demonstrated to reduce the FOP severity in cowpea roots in a size- and concentration-dependent manner. The three HMSNs at a low concentration of 100 mg/L, as well as HMSNs-19 at a high concentration of 1000 mg/L, were observed to have little effect on alleviating the disease incidence. HMSNs-406 were most effective at a concentration of 1000 mg/L, showing an up to 40.00% decline in the disease severity with significant growth-promoting effects on cowpea plants. Moreover, foliar application of HMSNs-406 (1000 mg/L) increased the salicylic acid (SA) content in cowpea roots by 4.3-fold, as well as the expression levels of SA marker genes ofPR-1(by 1.97-fold) andPR-5(by 9.38-fold), and its receptor gene ofNPR-1(by 1.62-fold), as compared with the FOP infected control plants. Meanwhile, another resistance-related gene ofPALwas also upregulated by 8.54-fold. Three defense-responsive enzymes of POD, PAL, and PPO were also involved in the HMSNs-enhanced disease resistance in cowpea roots, with varying degrees of reduction in activity. These results provide substantial evidence that HMSNs exert their Fusarium wilt suppression in cowpea plants by activating SA-dependent SAR (systemic acquired resistance) responses rather than directly suppressing FOP growth. Overall, for the first time, our results indicate a new role of HMSNs as a potent resistance inducer to serve as a low-cost, highly efficient, safe and sustainable alternative for plant disease protection.
研究领域:
农业
·
植物病理学
·
纳米技术
·
分子生物学
·
生物技术
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Stress Biology
·
2023
·Vol. 3
·期 1
·1-17
·
IF 0
Duan Kaili,Shen Qifang,Wang Yu,Xiang Ping,Shi Yutong,Yang Chenfei,Jiang Cong,Wang Guanghui,Xu Jin-Rong,Zhang Xue
Plant hormones are important for regulating growth, development, and plant-pathogen interactions. Some of them are inhibitory to growth of fungal pathogens but the underlying mechanism is not clear. In this study, we found that hyphal growth of Fusarium graminearum was significantly reduced by high concentrations of IAA and its metabolically stable analogue 2,4-dichlorophenoxyacetic acid (2,4-D). Besides inhibitory effects on growth rate, treatments with 2,4-D also caused significant reduction in conidiation, conidium germination, and germ tube growth. Treatments with 2,4-D had no obvious effect on sexual reproduction but significantly reduced TRI gene expression, toxisome formation, and DON production. More importantly, treatments with 2,4-D were inhibitory to infection structure formation and pathogenesis at concentrations higher than 100 µM. The presence of 1000 µM 2,4-D almost completely inhibited plant infection and invasive growth. In F. graminearum , 2,4-D induced ROS accumulation and FgHog1 activation but reduced the phosphorylation level of Gpmk1 MAP kinase. Metabolomics analysis showed that the accumulation of a number of metabolites such as glycerol and arabitol was increased by 2,4-D treatment in the wild type but not in the Fghog1 mutant. Transformants expressing the dominant active FgPBS2 S451D T455D allele were less sensitive to 2,4-D and had elevated levels of intracellular glycerol and arabitol induced by 2,4-D in PH-1. Taken together, our results showed that treatments with 2,4-D interfere with two important MAP kinase pathways and are inhibitory to hyphal growth, DON biosynthesis, and plant infection in F. graminearum .
研究领域:
植物病理学
·
分子生物学
·
植物激素
✓ 已验证
DOI
FUNGAL GENETICS AND BIOLOGY
·
2019
·
IF 3.314
Yinshan Zhang,Limin Wang,Shen Liang,Panpan Zhang,Ruijiao Kang,Mengjuan Zhang,Min Wang,Linlin Chen,Hongxia Yuan,Shengli Ding,Honglian Li
Histone deacetylases (HDACs) play essential roles in modulating chromatin structure to provide accessibility to gene regulators. Increasing evidence has linked HADCs to pathogenesis control in the filamentous plant fungi. However, its function remains unclear in Fusarium pseudograminearum, which has led to the emergence of the disease Fusarium crown rot in China. Here we identified the FpDEP1 gene, an orthologue of Saccharomyces cerevisiae DEP1 encoding a component of the Rpd3 histone deacetylase complex in F. pseudograminearum . The gene deletion mutant, ΔFpdep1, showed significantly retarded growth on PDA plates with reduced aerial hyphae formation. Pathogenicity tests displayed no typical leaf lesions and limited expansion capability of coleoptiles. Histopathological analysis indicated the ΔFpdep1 deletion mutant differentiated infectious hyphae and triggered massive reactive oxygen species (ROS) accumulation during the early infection stage, resulting in limited expansion to neighbor cells which was concurring with sensitivity to H 2 O 2 and SDS tests in vitro. FM4-64 staining revealed that the ΔFpdep1 deletion mutant was delayed in endocytosis. The FpDEP1-GFP transgene complemented the mutant phenotypes and the fusion protein co-localized with DAPI staining, indicating that the FpDEP1 gene product is localized to the nucleus in spores and mycelia. Immunoprecipitation coupled with LC-MS/MS and yeast two-hybrid screening identified the Rpd3L-like HDAC complex containing at least FpDep1, FpSds3, FpSin3, FpRpd3, FpRxt3, FpCti6, FpRho23, and FpUme6. These results suggest that FpDep1 is involved in a HDAC complex functioning on fungal development and pathogenesis in F. pseudograminearum .
研究领域:
真菌学
·
分子生物学
·
植物病理学
✓ 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）