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学者姓名：严芬
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Abstract ：
本研究旨在从类芽孢杆菌（Paenibacillus sp.）中克隆新型α-葡萄糖苷酶Aga432基因并通过定点突变提高α-葡萄糖苷酶Aga432的活性。从Paenibacillus sp.全基因组中获得表达α-葡萄糖苷酶的基因片段，进行序列分析，通过同源建模与分子对接并构建基因工程菌获得8株正向突变菌株，对重组Aga432和相对活性最高的正向突变体AT-2进行酶学性质研究，并探究重组α-葡萄糖苷酶Aga432、AT-2对生物膜的分散作用，评价其对小鼠胚胎成纤维细胞的毒性。结果表明，Aga432比活力为45.05 U/mg，突变体AT-2比活力为84.09 U/mg。与Aga432相比，AT-2的最适反应温度、最适反应pH改变并不明显，热稳定性有较大的提高，并且在酸性条件下较为稳定。突变体AT-2的Km为Aga432的1.87倍，Vmax值为Aga432的3.19倍，Kcat值为Aga432的2.33倍，Kcat/Km值为Aga432的1.07倍。体外细胞实验表明，15.0~30.0 μg/mL的Aga432、AT-2对细胞无明显毒性作用，具有良好的细胞相容性。生物膜分散作用结果表明，10.0~50.0 μg/mL浓度的两种重组α-葡萄糖苷酶对细菌生物膜具有显著的分散作用。本研究通过分子改造提升α-葡萄糖苷酶Aga432的热稳定性，为开发新型α-葡萄糖苷酶以及后续定向改造研究提供了基础和参考依据。
Keyword ：
α-葡萄糖苷酶 α-葡萄糖苷酶 分子对接 分子对接 同源建模 同源建模 生物膜分散作用 生物膜分散作用 酶学性质 酶学性质
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| GB/T 7714 | 路涵 , 方婷 , 牛晓旭 et al. 新型α-葡萄糖苷酶Aga432分子改造及酶学性质分析 [J]. | 食品工业科技 , 2025 , 46 (11) : 1-9 . |
| MLA | 路涵 et al. "新型α-葡萄糖苷酶Aga432分子改造及酶学性质分析" . | 食品工业科技 46 . 11 (2025) : 1-9 . |
| APA | 路涵 , 方婷 , 牛晓旭 , 翁晓敏 , 严芬 . 新型α-葡萄糖苷酶Aga432分子改造及酶学性质分析 . | 食品工业科技 , 2025 , 46 (11) , 1-9 . |
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目的：研究褐藻胶寡糖（alginate oligosaccharides,AOS）对葡聚糖硫酸钠（dextran sulfate sodium,DSS）诱导的小鼠炎症性肠病的影响，并通过AOS对肠道菌群和短链脂肪酸的影响探讨相关的可能机制。方法：雄性C57BL/6小鼠饮用5 d 1.5%DSS溶液进行急性结肠炎建模，同时以0.5、1 g/kg和1.5 g/kg的梯度剂量口服9 d AOS。在第10天评估体质量、结肠长度、疾病活动指数以及组织损伤。使用聚合酶链式反应法评估结肠中肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、核转录因子-κB(nuclear factor kappa B,NF-κB)、白细胞介素-6(interleukin-6,IL-6)、IL-1β、紧密连接蛋白（zonal occludens-1,ZO-1）和G蛋白偶联受体43(G protein-coupled receptor43,GPR43)的mRNA表达水平。采用高效液相色谱测定短链脂肪酸含量并分别对微生物群落进行测序。结果：AOS显著减轻了结肠炎症状，如体质量下降、结肠缩短、组织损伤，并降低了TNF-α、NF-κB、IL-6和IL-1β的mRNA表达，增加了ZO-1和GPR43的mRNA表达。且AOS可显著增加丙酸杆菌的数量，同时抑制肠球菌和放线菌的生长；高剂量AOS还可以促进乳酸、乙酸和丙酸的产生。结论：AOS对DSS诱导的小鼠结肠炎具有积极的改善作用，其作用机制可能与肠道菌群和短链脂肪酸的调节有关，具有作为改善肠道炎症的功能性食品的潜力。
Keyword ：
炎症性肠病 炎症性肠病 短链脂肪酸 短链脂肪酸 肠道菌群 肠道菌群 褐藻胶寡糖 褐藻胶寡糖
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| GB/T 7714 | 任晓敏 , 秦明珍 , 张少龙 et al. 褐藻胶寡糖调节肠道菌群稳态及短链脂肪酸代谢并改善炎症性肠病 [J]. | 食品科学 , 2025 , 46 (15) : 224-231 . |
| MLA | 任晓敏 et al. "褐藻胶寡糖调节肠道菌群稳态及短链脂肪酸代谢并改善炎症性肠病" . | 食品科学 46 . 15 (2025) : 224-231 . |
| APA | 任晓敏 , 秦明珍 , 张少龙 , 张帆 , 严芬 . 褐藻胶寡糖调节肠道菌群稳态及短链脂肪酸代谢并改善炎症性肠病 . | 食品科学 , 2025 , 46 (15) , 224-231 . |
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从海洋细菌Zobellia sp.B2中克隆新型纤维素酶CelL7基因,同时添加碳水化合物结合模块家族3(carbohydrate-binding module family 3,CBM3)基因构建融合基因CelL7-CBM3,并实现其编码的融合蛋白CelL7-CBM3在大肠杆菌BL21中实现异源表达,利用亲和层析柱获得纯化蛋白.CelL7基因序列全长1 077 bp,编码358 个氨基酸残基,理论蛋白分子质量为40.39 kDa.CelL7和CelL7-CBM3的比酶活力分别为2 249.81 U/mg和2 915.75 U/mg.CelL7与CelL7-CBM3的最适反应温度均为50℃,最适pH值分别为5.0和5.5,Mn2+和Fe2+能激活CelL7,Cu2+抑制CelL7的活力,CelL7可降解羧甲基纤维素钠、纤维二糖和木聚糖.以羧甲基纤维素钠为底物时,CelL7-CBM3的米氏常数(Km)为11.70 mg/mL,较CelL7的Km(13.23 mg/mL)有所降低,表明添加结合结构域后的融合酶对羧甲基纤维素钠的亲和力增强;最大反应速率(Vmax)为175.44 mg/(mL·min),催化常数(Kcat)为2.78 s-1,Kcat/Km为0.24 mL/(mg·s),与CelL7相比变化不大.生物膜清除实验表明,10.0～60.0 μg/mL的CelL7和30.0～60.0 μg/mL CelL7-CBM3能够有效分散生物膜,减少生物膜量.
Keyword ：
异源表达 异源表达 生物膜清除 生物膜清除 纤维素酶 纤维素酶 结构域融合 结构域融合
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| GB/T 7714 | 翁晓敏 , 胡诗琦 , 蔡佳琪 et al. 海洋来源纤维素酶CelL7的异源表达、酶学表征及生物膜清除作用 [J]. | 食品科学 , 2025 , 46 (6) : 124-132 . |
| MLA | 翁晓敏 et al. "海洋来源纤维素酶CelL7的异源表达、酶学表征及生物膜清除作用" . | 食品科学 46 . 6 (2025) : 124-132 . |
| APA | 翁晓敏 , 胡诗琦 , 蔡佳琪 , 洪健渠 , 严芬 . 海洋来源纤维素酶CelL7的异源表达、酶学表征及生物膜清除作用 . | 食品科学 , 2025 , 46 (6) , 124-132 . |
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The rise of antibiotic-resistant bacteria poses a serious global health threat, highlighting the urgent need for novel strategies beyond conventional antibiotic therapies. This study explores the potential of microbe-imprinted polymers (MIPs) as innovative, pathogen-specific affinity agents. Utilizing microbial surface-initiated polymerization, MIPs are in-situ synthesized on the surface of target microbes, creating flexible heteropolymers that precisely replicate microbial surface structures. This method exhibits high affinity (KD = 2.7x108 CFU/mL for E. coli) and selectivity at the strain level. MIPs offer significant advantages over traditional antibodies, including greater stability, cost-effectiveness, and a broader spectrum of binding capabilities, making them effective for identifying and targeting various microbial strains, including unidentified or drug-resistant variants. Moreover, their favorable biocompatibility and functional resilience in diverse environments position MIPs as promising candidates for rapid pathogen detection and therapeutic applications. This research paves the way for advanced biomimetic materials in microbe-specific diagnostics and combating infections, addressing the critical need for effective tools in antibiotic resistance surveillance.
Keyword ：
Affinity Affinity Antibiotic resistance Antibiotic resistance Antibody mimics Antibody mimics Microbe-imprinted polymers Microbe-imprinted polymers Microbial recognition Microbial recognition
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| GB/T 7714 | Wu, Yuanzi , Zhou, Kaiqiang , Li, Wenhui et al. Microbe-imprinted polymers for rapid drug-resistant bacteria recognition [J]. | CHEMICAL ENGINEERING JOURNAL , 2025 , 512 . |
| MLA | Wu, Yuanzi et al. "Microbe-imprinted polymers for rapid drug-resistant bacteria recognition" . | CHEMICAL ENGINEERING JOURNAL 512 (2025) . |
| APA | Wu, Yuanzi , Zhou, Kaiqiang , Li, Wenhui , Huan, Min , Yu, Zhichao , Yan, Fen et al. Microbe-imprinted polymers for rapid drug-resistant bacteria recognition . | CHEMICAL ENGINEERING JOURNAL , 2025 , 512 . |
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This study aimed to clone the novel α-glucosidase gene Aga432 from Paenibacillus sp. and enhance its catalytic activity through site-directed mutagenesis. A gene fragment encoding α-glucosidase was successfully amplified from the genomic DNA of Paenibacillus sp., comprehensive sequence analysis was performed, and homology modeling and molecular docking were employed to construct gene-engineered strains. Eight positive mutant strains were identified, among which the enzymatic properties of recombinant Aga432 and the highest relative activity mutant AT-2 were characterized. Additionally, the dispersing effects of recombinant α-glucosidases Aga432 and AT-2 on biofilms were explored, and their toxicity to mouse embryo fibroblasts was evaluated. The results revealed that the specific activity of Aga432 was 45.05 U/mg, while the mutant AT-2 exhibited a significantly enhanced specific activity of 84.09 U/mg. Although the optimal reaction temperature and pH for AT-2 were essentially unaltered relative to Aga432, its thermal stability was significantly enhanced, and it exhibited heightened stability under acidic conditions. The Km of mutant AT-2 was 2.18 times that of Aga432, the Vmax was 3.19 times, the Kcat was 2.33 times, and the Kcat/Km was 1.07 times that of Aga432. In vitro cellular assays indicated that Aga432 and AT-2 at concentrations of 15.0~30.0 μg/mL were non-toxic and exhibited good cell compatibility. Biofilm dispersal assays demonstrated that both recombinant α-glucosidases at concentrations ranging from 10.0 to 50.0 μg/mL significantly dispersed bacterial biofilms (P © The Author(s) 2025.
Keyword ：
Bioassay Bioassay Biofilms Biofilms Gene encoding Gene encoding
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| GB/T 7714 | Lu, Han , Fang, Ting , Niu, Xiaoxu et al. Molecular Modification and Enzymatic Properties of the Novel α-Glucosidase Aga432 [J]. | Science and Technology of Food Industry , 2025 , 46 (11) : 185-193 . |
| MLA | Lu, Han et al. "Molecular Modification and Enzymatic Properties of the Novel α-Glucosidase Aga432" . | Science and Technology of Food Industry 46 . 11 (2025) : 185-193 . |
| APA | Lu, Han , Fang, Ting , Niu, Xiaoxu , Weng, Xiaomin , Yan, Fen . Molecular Modification and Enzymatic Properties of the Novel α-Glucosidase Aga432 . | Science and Technology of Food Industry , 2025 , 46 (11) , 185-193 . |
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Non-alcoholic fatty liver disease (NAFLD) has become a globally rising issue that can cause liver-related morbidity and mortality. Recent studies have revealed that a high-fat diet (HFD) highly contributes to the prevalence and progression of NAFLD via impacting gut microbiota and lipid metabolism signal pathways. Resveratrol (RSV), a natural bioactive compound, has exhibited potential for preventing and alleviating NAFLD. However, due to the poor bioavailability of RSV, its strengths and underlying mechanisms for NAFLD therapeutic potential are poorly understood. To address this, we utilized Hydroxypropyl-beta-Cyclodextrin (HBC) to encapsulate RSV to enhance its water solubility and conducted prevention and intervention experiments in HFDfed mice. The results showed that the HBC has significantly enhanced the water solubility of RSV by 250-fold and the HBC-RSV better prevented and alleviated the liver steatosis, obesity and abnormal lipid metabolism induced by HFD than RSV alone. Meanwhile, combining HFD and HBC-RSV or RSV prevented HFD mice progressing to NAFLD. Besides, further investigation indicated that RSV could resist liver injury and obesity by modulating gut microbiota, raising the levels of short-chain fatty acids (SCFAs) and activating AMP-activated protein kinase (AMPK) signaling pathway. The activated pathway down-regulated sterol receptor element binding protein 1c (SREBP1c) and acetyl-coenzyme A carboxylase (ACC) to decrease lipid synthesis and up-regulated peroxisome proliferators-activated receptor & alpha; (PPAR & alpha;) to promote the fatty acid oxidation, thus preventing NAFLD. Our findings suggested that water solubility-enhanced RSV beneficially modulated gut microbiota, altered gut microbiota-derived SCFAs, and activated lipid metabolism regulatory pathways, providing potential for NAFLD prevention and alleviation.
Keyword ：
AMPK AMPK Gut microbiota Gut microbiota Hydroxypropyl-beta-Cyclodextrin Hydroxypropyl-beta-Cyclodextrin NAFLD NAFLD resveratrol resveratrol SCFAs SCFAs
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| GB/T 7714 | Ke, Wenya , Huang, Juan , Zhong, Yi et al. Hydroxypropyl-beta-Cyclodextrin embedded resveratrol regulates gut microbiota to prevent NAFLD via activating AMPK signaling pathway [J]. | FOOD BIOSCIENCE , 2023 , 54 . |
| MLA | Ke, Wenya et al. "Hydroxypropyl-beta-Cyclodextrin embedded resveratrol regulates gut microbiota to prevent NAFLD via activating AMPK signaling pathway" . | FOOD BIOSCIENCE 54 (2023) . |
| APA | Ke, Wenya , Huang, Juan , Zhong, Yi , Shi, Yuhong , Yan, Fen , Huang, Da et al. Hydroxypropyl-beta-Cyclodextrin embedded resveratrol regulates gut microbiota to prevent NAFLD via activating AMPK signaling pathway . | FOOD BIOSCIENCE , 2023 , 54 . |
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Bacterial infection of wounds is one of great concern to patients, and rapid and correct detection of bacterial infection is crucial to ensure accurate diagnosis and early intervention. Based on the principle that glucose can only be consumed by live bacteria, a fluorescent biosensor was constructed to detect four kinds of common infectious bacteria (Acinetobacter baumannii, Escherichia coli, Pseudomonas aeruginosa and Staphylococcus aureus) in skin wounds, taking advantage of magnetic bead-aptamer for recognizing, sorting and enrichment, platinum nanoparticles for signal amplification. The linear detection range of AB, EC, PA and SA were 27- 8 x 106 CFU/mL, 10- 2.5 x 107 CFU/mL, 34- 2.5 x 107 CFU/mL and 10- 1.0 x 107 CFU/mL, respectively, and the limits of detection were 27 CFU/mL, 10 CFU/mL, 34 CFU/mL and 2 CFU/mL. Furthermore, all target bacteria in samples containing 8 x 108 CFU/mL of other interfering bacteria have been successfully identified and quantified. The proposed method was also successfully applied to the detection of bacterial infection in skin wounds of mouse and human, including the detection of separate bacterial infection as well as a coinfection. The recovery of this method was in the range of 90.161- 109.961%. Thus, this proposed method can be a promising candidate for rapid and convenient evaluation of infectious bacteria in point-of-care settings.
Keyword ：
Aptamer Aptamer Bacterial infection Bacterial infection Fluorescent biosensor Fluorescent biosensor Platinum nanoparticles Platinum nanoparticles Wound Wound
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| GB/T 7714 | Gao, Lanmei , Zheng, Houbing , Hu, Yuanlong et al. Fluorescent biosensor based on MB-APT combined with Pt NPs for the detection of infectious bacteria in mouse and human wounds [J]. | SENSORS AND ACTUATORS B-CHEMICAL , 2023 , 393 . |
| MLA | Gao, Lanmei et al. "Fluorescent biosensor based on MB-APT combined with Pt NPs for the detection of infectious bacteria in mouse and human wounds" . | SENSORS AND ACTUATORS B-CHEMICAL 393 (2023) . |
| APA | Gao, Lanmei , Zheng, Houbing , Hu, Yuanlong , Zhong, Yi , Jiang, Linhai , Wu, Yuanzi et al. Fluorescent biosensor based on MB-APT combined with Pt NPs for the detection of infectious bacteria in mouse and human wounds . | SENSORS AND ACTUATORS B-CHEMICAL , 2023 , 393 . |
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In view of the biological significance and micro-heterogeneity of protein glycosylation for human health, specific enrichment of N-glycosylated proteins/peptides from complex biological samples is a prerequisite for the discovery of disease biomarkers and clinical diagnosis. In this work, we propose a "grafting-from" N-glycoprotein enriching method based on the in-situ growth of thermoresponsive polymer brushes from the N-glycosylated site of proteins. The initiator was first attached to the pre-oxidized glycan moieties by hydrazide chemistry, from which the thermoresponsive polymers can be grown to form giant protein-polymer conjugates (PPC). The thermosensitive PPC can be precipitated and separated by raising the temperature to above its lower critical solubility temperature (LCST). Mass spectrometry verified 210 N-glycopeptides corresponding to 136 N-glycoproteins in the rabbit serum. These results demonstrate the capability of the tandem thermoprecipitation strategy to enrich and separate N-glycoprotein/glycopeptide. Due to its simplicity and efficiency specifically, this method holds the potential for identifying biomarkers from biological samples in N-glycoproteome analysis.
Keyword ：
Glycan-selective Glycan-selective glycopeptide glycopeptide N-Glycoprotein N-Glycoprotein Thermoprecipitation Thermoprecipitation
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| GB/T 7714 | Shu, Jingjing , Xiong, Wenli , Zhang, Ran et al. Glycan-selective in-situ growth of thermoresponsive polymers for thermoprecipitation and enrichment of N-glycoprotein/glycopeptides [J]. | TALANTA , 2023 , 253 . |
| MLA | Shu, Jingjing et al. "Glycan-selective in-situ growth of thermoresponsive polymers for thermoprecipitation and enrichment of N-glycoprotein/glycopeptides" . | TALANTA 253 (2023) . |
| APA | Shu, Jingjing , Xiong, Wenli , Zhang, Ran , Ma, Shanyun , Zhou, Kaiqiang , Wang, Xuwei et al. Glycan-selective in-situ growth of thermoresponsive polymers for thermoprecipitation and enrichment of N-glycoprotein/glycopeptides . | TALANTA , 2023 , 253 . |
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本发明涉及一种重组褐藻胶裂解酶AlyL7及其应用，所述褐藻胶裂解酶的氨基酸序列如SEQ ID NO.1所示，其编码基因的核苷酸序列如SEQ ID NO.2所示。所述重组褐藻胶裂解酶属于双功能型褐藻胶裂解酶，其最适反应温度为40℃，最适反应pH为9.0，在0‑25℃、pH 5.0‑10.0的条件时较稳定，对海藻酸钠、聚古洛糖醛酸、聚甘露糖醛酸均具有降解活性。所述重组褐藻胶裂解酶也属于内切酶，其酶解海藻酸钠的终产物包括不饱和1‑5糖，以不饱和2‑3糖为主。所述褐藻胶裂解酶反应速度快，效率高，具有良好的工业化应用潜质。
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| GB/T 7714 | 严芬 , 陈骏颖 , 钟金福 et al. 一种重组褐藻胶裂解酶AlyL7及其应用 : CN202111240122.4[P]. | 2021-10-25 00:00:00 . |
| MLA | 严芬 et al. "一种重组褐藻胶裂解酶AlyL7及其应用" : CN202111240122.4. | 2021-10-25 00:00:00 . |
| APA | 严芬 , 陈骏颖 , 钟金福 , 张少龙 . 一种重组褐藻胶裂解酶AlyL7及其应用 : CN202111240122.4. | 2021-10-25 00:00:00 . |
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本发明涉及一种忧遁草抗氧化十三肽及其制备方法和应用，属于生物技术领域。所述抗氧化肽氨基酸序列为LLPENDPSANHLM，该抗氧化肽的制备方法是以忧遁草叶为原料，采用碱溶酸沉淀法提取粗制品，再经分离纯化、鉴定及合成技术得到所述抗氧化肽。药理实验证明，本发明提供的忧遁草抗氧化十三肽能在不同程度上保护HepG‑2细胞免受H2O2毒害，其具有结构简单、抗氧化活力强、安全性高的特点，可以作为化学合成抗氧化剂的优良替代。本发明也为食品、药品及化妆品行业研发新型天然添加剂提供了理论依据和实践参考。
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| GB/T 7714 | 严芬 , 刘文静 , 张少龙 et al. 一种忧遁草抗氧化十三肽及其制备方法和应用 : CN202210288893.9[P]. | 2022-03-23 00:00:00 . |
| MLA | 严芬 et al. "一种忧遁草抗氧化十三肽及其制备方法和应用" : CN202210288893.9. | 2022-03-23 00:00:00 . |
| APA | 严芬 , 刘文静 , 张少龙 , 费庆彬 . 一种忧遁草抗氧化十三肽及其制备方法和应用 : CN202210288893.9. | 2022-03-23 00:00:00 . |
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