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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
期刊论文 | 2025 , 46 (6) , 124-132 | 食品科学
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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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新型α-葡萄糖苷酶Aga432分子改造及酶学性质分析
期刊论文 | 2025 , 1-9 | 食品工业科技
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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 : 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 (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 , 1-9 .
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Microbe-imprinted polymers for rapid drug-resistant bacteria recognition Scopus
期刊论文 | 2025 , 512 | Chemical Engineering Journal
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Abstract :

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 = 2.7×108 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. © 2025

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, Y. , Zhou, K. , Li, W. et al. Microbe-imprinted polymers for rapid drug-resistant bacteria recognition [J]. | Chemical Engineering Journal , 2025 , 512 .
MLA Wu, Y. et al. "Microbe-imprinted polymers for rapid drug-resistant bacteria recognition" . | Chemical Engineering Journal 512 (2025) .
APA Wu, Y. , Zhou, K. , Li, W. , Huan, M. , Yu, Z. , Yan, F. et al. Microbe-imprinted polymers for rapid drug-resistant bacteria recognition . | Chemical Engineering Journal , 2025 , 512 .
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Glycan-selective in-situ growth of thermoresponsive polymers for thermoprecipitation and enrichment of N-glycoprotein/glycopeptides SCIE
期刊论文 | 2023 , 253 | TALANTA
WoS CC Cited Count: 3
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Abstract :

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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Glycan-selective in-situ growth of thermoresponsive polymers for thermoprecipitation and enrichment of N-glycoprotein/glycopeptides Scopus
期刊论文 | 2023 , 253 | Talanta
Glycan-selective in-situ growth of thermoresponsive polymers for thermoprecipitation and enrichment of N-glycoprotein/glycopeptides EI
期刊论文 | 2023 , 253 | Talanta
Hydroxypropyl-beta-Cyclodextrin embedded resveratrol regulates gut microbiota to prevent NAFLD via activating AMPK signaling pathway SCIE
期刊论文 | 2023 , 54 | FOOD BIOSCIENCE
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Abstract :

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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Hydroxypropyl-beta-Cyclodextrin embedded resveratrol regulates gut microbiota to prevent NAFLD via activating AMPK signaling pathway Scopus
期刊论文 | 2023 , 54 | Food Bioscience
Fluorescent biosensor based on MB-APT combined with Pt NPs for the detection of infectious bacteria in mouse and human wounds SCIE
期刊论文 | 2023 , 393 | SENSORS AND ACTUATORS B-CHEMICAL
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Abstract :

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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Fluorescent biosensor based on MB-APT combined with Pt NPs for the detection of infectious bacteria in mouse and human wounds EI
期刊论文 | 2023 , 393 | Sensors and Actuators B: Chemical
Fluorescent biosensor based on MB-APT combined with Pt NPs for the detection of infectious bacteria in mouse and human wounds Scopus
期刊论文 | 2023 , 393 | Sensors and Actuators B: Chemical
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 incoPat
专利 | 2022-04-28 00:00:00 | CN202210458985.7
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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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抑制香蕉采后病害的贝莱斯芽孢杆菌LZT14及其应用 incoPat
专利 | 2021-12-23 00:00:00 | CN202111586238.3
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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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一株控制香蕉采后病害的酵母菌P4及其使用方法 incoPat
专利 | 2021-08-31 00:00:00 | CN202111008228.1
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本发明公开一株用于香蕉采后病害的酵母菌P4及其使用方法。所述酵母菌P4在中国微生物菌种保藏管理委员会普通微生物中心的保藏编号为CGMCC No.22561。所述酵母菌P4的使用方法为:菌株活化,菌体用无菌水配成1×108 cells/mL的菌悬液;将菌悬液喷洒于香蕉表面,自然晾干后,放入0.015 mm厚的聚乙烯薄膜袋中,恒温恒湿贮藏。所述酵母菌P4可以控制香蕉炭疽病,同时该菌株抑菌谱较广,可以控制黄曲霉、拟盘多毛孢属、枇杷胶孢炭疽菌、香蕉镰刀菌等。

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GB/T 7714 严芬 , 吴芸芸 , 蔡婷 et al. 一株控制香蕉采后病害的酵母菌P4及其使用方法 : CN202111008228.1[P]. | 2021-08-31 00:00:00 .
MLA 严芬 et al. "一株控制香蕉采后病害的酵母菌P4及其使用方法" : CN202111008228.1. | 2021-08-31 00:00:00 .
APA 严芬 , 吴芸芸 , 蔡婷 , 刘文静 , 张帆 . 一株控制香蕉采后病害的酵母菌P4及其使用方法 : CN202111008228.1. | 2021-08-31 00:00:00 .
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Unimolecular Cascaded Multienzyme Conjugates Modulate the Microenvironment of Diabetic Wound to Promote Healing SCIE
期刊论文 | 2023 , 25 (1) , 43-54 | BIOMACROMOLECULES
WoS CC Cited Count: 2
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An abnormal microenvironment underlies poor healing in chronic diabetic chronic wounds. However, effectively modulating the microenvironment of the diabetic wound remains a great challenge due to sustained oxidative stress and chronic inflammation. Here, we present a unimolecular enzyme-polymer conjugate that demonstrates excellent multienzymatic cascade activities. The cascaded enzyme conjugates (CECs) were synthesized by grafting poly(N-acryloyl-lysine) (pLAAm) from the glycan moieties of glucose oxidase (GOx) via glycan-initiated polymerization. The resulting CECs exhibited multiple enzymatic properties of GOx, superoxide dismutase mimic, and catalase mimic activities simultaneously. The CECs facilitated the depletion of high blood glucose, ROS scavenging, bacteria-killing, anti-inflammatory effects, and sustained oxygen generation, which restored the microenvironment in diabetic wounds. In vivo results from a diabetic mouse model confirmed the capacity and efficiency of the cascade reaction for diabetic wound healing. Our findings demonstrate that the three-in-one enzyme-polymer conjugates alone can modulate the diabetic microenvironment for wound healing.

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GB/T 7714 Wu, Yuanzi , Lyu, Yanwei , Li, Ling et al. Unimolecular Cascaded Multienzyme Conjugates Modulate the Microenvironment of Diabetic Wound to Promote Healing [J]. | BIOMACROMOLECULES , 2023 , 25 (1) : 43-54 .
MLA Wu, Yuanzi et al. "Unimolecular Cascaded Multienzyme Conjugates Modulate the Microenvironment of Diabetic Wound to Promote Healing" . | BIOMACROMOLECULES 25 . 1 (2023) : 43-54 .
APA Wu, Yuanzi , Lyu, Yanwei , Li, Ling , Zhou, Kaiqiang , Cai, Jingwen , Wang, Xuwei et al. Unimolecular Cascaded Multienzyme Conjugates Modulate the Microenvironment of Diabetic Wound to Promote Healing . | BIOMACROMOLECULES , 2023 , 25 (1) , 43-54 .
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Unimolecular Cascaded Multienzyme Conjugates Modulate the Microenvironment of Diabetic Wound to Promote Healing EI
期刊论文 | 2024 , 25 (1) , 43-54 | Biomacromolecules
Unimolecular Cascaded Multienzyme Conjugates Modulate the Microenvironment of Diabetic Wound to Promote Healing Scopus
期刊论文 | 2024 , 25 (1) , 43-54 | Biomacromolecules
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