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学者姓名：严芬
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Abstract ：
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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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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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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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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本发明涉及一种具有抗炎抗氧化活性的忧遁草多肽及其制备方法和应用，属于生物技术领域。所述忧遁草多肽序列的氨基酸序列为DKFEDNLYSAH，分子量为1338.2 Da。所述忧遁草多肽是以忧遁草为原料，采用超声辅助碱溶酸沉法获取忧遁草粗蛋白，再经分离纯化、鉴定、Fmoc固相合成得到的。试验证明，本发明所述忧遁草多肽具有很好的抗炎活性及抗氧化活性，具有很好的应用潜力和价值。
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| GB/T 7714 | 严芬 , 刘文静 , 刘聪 et al. 一种具有抗炎抗氧化活性的忧遁草多肽及其制备方法和应用 : CN202210458985.7[P]. | 2022-04-28 00:00:00 . |
| MLA | 严芬 et al. "一种具有抗炎抗氧化活性的忧遁草多肽及其制备方法和应用" : CN202210458985.7. | 2022-04-28 00:00:00 . |
| APA | 严芬 , 刘文静 , 刘聪 , 陈晓杰 , 张帆 . 一种具有抗炎抗氧化活性的忧遁草多肽及其制备方法和应用 : CN202210458985.7. | 2022-04-28 00:00:00 . |
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本发明公开了一株抑制香蕉采后病害的贝莱斯芽孢杆菌LZT14及其应用，属于植物采后病害防治技术领域。所述菌株从福州大学荔枝树周围土壤样品中分离筛选得到，已于2021年8月16日保藏在中国微生物菌种保藏管理委员会普通微生物中心，保藏编号为CGMCC No.23150。该菌株抑菌谱广、且可应用于香蕉采后病害预防和贮藏保鲜。该菌株的使用方法为：菌株活化，洗涤后重悬；将菌悬液喷洒于香蕉表面，风干，放入0.015 mm厚的聚乙烯薄膜袋中，恒温恒湿贮藏。香蕉体内试验结果表明，在采后28℃，80%相对湿度环境中储藏时，该菌株可以明显降低香蕉的病情指数，生防应用前景良好。
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| GB/T 7714 | 严芬 , 刘聪 , 陈晓杰 et al. 抑制香蕉采后病害的贝莱斯芽孢杆菌LZT14及其应用 : CN202111586238.3[P]. | 2021-12-23 00:00:00 . |
| MLA | 严芬 et al. "抑制香蕉采后病害的贝莱斯芽孢杆菌LZT14及其应用" : CN202111586238.3. | 2021-12-23 00:00:00 . |
| APA | 严芬 , 刘聪 , 陈晓杰 , 吴崟沣 , 方婷 . 抑制香蕉采后病害的贝莱斯芽孢杆菌LZT14及其应用 : CN202111586238.3. | 2021-12-23 00:00:00 . |
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本专利公开了一种忧遁草的抗氧化十一肽及其制备方法和应用，所述抗氧化肽序列为DMGPPLSEKLH。本发明以忧遁草为原料，采用超声辅助碱溶酸提取技术进行制备，进一步通过离子色谱、反相高效液相色谱等分离纯化技术建立忧遁草抗氧化肽的纯化方法。体外抗氧化实验表明，所述忧遁草抗氧化十一肽可以有效清除DPPH、ABTS、羟基自由基等自由基，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，而且具有一定的抗肿瘤活性，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 , 张丹 et al. 一种忧遁草抗氧化十一肽及其制备方法和应用 : CN202210103859.X[P]. | 2022-01-28 00:00:00 . |
| MLA | 严芬 et al. "一种忧遁草抗氧化十一肽及其制备方法和应用" : CN202210103859.X. | 2022-01-28 00:00:00 . |
| APA | 严芬 , 刘文静 , 张丹 , 胡诗琦 , 吴崟沣 . 一种忧遁草抗氧化十一肽及其制备方法和应用 : CN202210103859.X. | 2022-01-28 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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