Journal of biotechnology最新文献_第3页

Cell specific nutrient feeding in perfusion mode enhances hMSC growth in stirred tank bioreactor process 在搅拌槽生物反应器中，灌注模式下细胞特异性营养物的饲喂可促进hMSC的生长。

IF 3.9 2区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Journal of biotechnology

Pub Date : 2025-12-15 DOI: 10.1016/j.jbiotec.2025.12.006

Johanna Pechan , Kristina Engström , Nora Mac Key , Lina Sörvik , Véronique Chotteau

Human Mesenchymal Stromal Cells (hMSCs) are a safe option for allogeneic cell therapy across various diseases, but their manufacturing process requires improvement to broaden accessibility. In this study, a state-of-the-art planar multi-vessel process was transferred to a stirred tank bioreactor using microcarriers to support the growth of adherent cells. The frequent medium exchange strategy from planar culture guided the design of the bioreactor process. However, complete medium changes in the bioreactor resulted in limited cell expansion and higher glucose consumption compared to planar culture. To enhance expansion, a 0.1 L perfusion bioreactor was tested, enabling continuous medium exchange. Three perfusion approaches were evaluated: (1) maintaining a target cell-specific glucose consumption rate, (2) varying the perfusion rate, and (3) applying a cell-specific perfusion rate. Implementing targeted glucose feeding (TAFE) reduced lactate production, while increasing perfusion rates improved cell density. The highest expansion was achieved using a cell-specific perfusion rate of 5 nL cell⁻¹ day⁻¹ combined with a target glucose consumption rate (q_glc) of 15 pmol cell⁻¹ day⁻¹, resulting in a 5.4-fold higher expansion factor than daily medium changes in stirred tank bioreactors. This optimized process represents a key advancement toward producing clinically relevant quantities of hMSCs.

人间充质基质细胞（hMSCs）是异体细胞治疗多种疾病的安全选择，但其制造工艺需要改进以扩大可及性。在本研究中，将最先进的平面多容器工艺转移到搅拌槽生物反应器中，使用微载体支持贴壁细胞的生长。平面培养的频繁介质交换策略指导了生物反应器工艺的设计。然而，与平面培养相比，生物反应器中完全改变培养基导致细胞扩增有限，葡萄糖消耗更高。为了增强膨胀，我们测试了0.1L的灌注生物反应器，以实现连续的介质交换。评估了三种灌注方法：(1)维持靶细胞特异性葡萄糖消耗率，(2)改变灌注率，(3)应用细胞特异性灌注率。实施靶向葡萄糖喂养（TAFE）减少乳酸生成，而增加灌注率提高细胞密度。当细胞特异性灌注率为5 nL细胞-1天-1，目标葡萄糖消耗率（qglc）为15 pmol细胞-1天-1时，达到了最高的扩增率，其扩增系数比搅拌槽生物反应器中每日培养基变化高5.4倍。这个优化的过程代表了生产临床相关数量的hMSCs的关键进展。

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引用次数: 0

Enhancing oxidative stress defense to improve docosahexaenoic acid production in Schizochytrium sp. HX-308 增强氧化应激防御提高Schizochytrium sp. HX-308二十二碳六烯酸产量

IF 3.9 2区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Journal of biotechnology

Pub Date : 2025-12-12 DOI: 10.1016/j.jbiotec.2025.12.005

Chunxiao Yan , Shuai Zhang , Zhihao Huang , Fei Du , Qian Xu , Wang Ma , Guang Yang , Xiaoman Sun

As an excellent host for docosahexaenoic acid (DHA) biosynthesis, Schizochytrium sp. still faces inherent limitations in DHA yield due to oxidative damage induced by reactive oxygen species (ROS). To effectively reduce the ROS levels, the optimal combination of mixed antioxidants (52.95 mg L⁻¹ phytic acid, 32.00 mg L⁻¹ licorice extract, 36.28 mg L⁻¹ ascorbic acid palmitate) was selected by single factor experiment combined with response surface optimization. The DHA titer increased from 24.0 g L⁻¹ to 33.5 g L⁻¹. However, exogenous addition of antioxidants results in increased the cost of DHA production. Enhancing the intracellular antioxidant capacity through gene engineering has become another feasible strategy. Transcriptome analysis indicated that the transcriptional levels of antioxidant-related genes SOD1 (superoxide dismutase), APX5 (ascorbic acid peroxidase), and GST3 (glutathione S-transferase) were significantly upregulated during DHA biosynthesis. Then, the engineered strain SOD1-APX5-GST3–2 was constructed by overexpressing the three genes, and the DHA and lipid titers were 33.1 g L⁻¹ and 64.1 g L⁻¹, respectively. Scale-up in a 5 L bioreactor obtained 32.5 g L⁻¹ DHA and 63.5 g L⁻¹ lipid, and the ROS level decreased by 30.5 %, demonstrating engineered strain's stability and laying the foundation for industrial-scale production. This study verified that reducing ROS levels promoted DHA biosynthesis in Schizochytrium sp. from the two dimensions of exogenous antioxidants and gene engineering, which provided novel insights for industrial lipid production.

Schizochytrium sp.作为二十二碳六烯酸（二十二碳六烯酸，DHA）生物合成的优良宿主，由于活性氧（reactive oxygen species， ROS）诱导的氧化损伤，其DHA产量仍存在固有的局限性。为有效降低ROS水平，通过单因素试验结合响应面优化，筛选出混合抗氧化剂（52.95mg -1植酸、32.00mg -1甘草提取物、36.28mg -1抗坏血酸棕榈酸酯）的最佳组合。DHA滴度由24.0gL-1提高到33.5gL-1。然而，外源添加抗氧化剂会导致DHA生产成本的增加。通过基因工程增强细胞内抗氧化能力已成为另一种可行的策略。转录组分析表明，抗氧化相关基因SOD1（超氧化物歧化酶）、APX5（抗坏血酸过氧化物酶）和GST3（谷胱甘肽s -转移酶）的转录水平在DHA生物合成过程中显著上调。通过过表达这3个基因构建工程菌株SOD1-APX5-GST3-2，其DHA滴度为33.1gL-1，脂质滴度为64.1gL-1。在5L生物反应器中放大后，得到了32.5gL-1 DHA和63.5gL-1脂质，ROS水平下降了30.5%，证明了工程菌株的稳定性，为工业规模生产奠定了基础。本研究从外源抗氧化剂和基因工程两个维度验证了ROS水平的降低促进了Schizochytrium sp. DHA的生物合成，为工业脂质生产提供了新的见解。

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引用次数: 0

Next level of p-phenylene diisothiocyanate-based covalent immobilization of β-D-galactosidase: Technical optimization as an application 基于对苯二异硫氰酸酯的β-d-半乳糖苷酶共价固定化的新水平：技术优化应用。

IF 3.9 2区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Journal of biotechnology

Pub Date : 2025-12-10 DOI: 10.1016/j.jbiotec.2025.12.004

Tabea L. Boehme , Bernadette Straub , Anja Oberle , Ursula Eschenhagen , Magnus S. Schmidt

In this study, a continuous lactose hydrolysis process in a fixed-bed reactor was developed using β-galactosidase covalently immobilized on resin beads via 1,4-phenylenediisothiocyanate (PDC) as linker. Process conditions, including temperature, enzyme loading, perfusion speed, and repeated perfusion of the same substrate solution were systematically varied. The highest glucose yields were obtained at 55 °C, with increased yields observed at low perfusion speeds, high enzyme loadings, and upon repeated perfusions. Under optimized cycle perfusion over 72 h, final lactose conversion reached approximately 90 % at 37 °C and 80 % at 22 °C. A hydrolysis process in a fixed-bed reactor was successfully established, although further optimization is required.

本研究以1,4-苯二异硫氰酸酯（PDC）为连接剂，将β-半乳糖苷酶共价固定在树脂珠上，建立了固定床反应器中乳糖连续水解工艺。工艺条件，包括温度，酶载量，灌注速度，重复灌注相同的底物溶液系统地改变。在55°C时获得最高葡萄糖产量，在低灌注速度、高酶负荷和重复灌注时观察到产量增加。在优化循环灌注超过72h的情况下，最终乳糖转化率在37°C时达到约90%，在22°C时达到80%。在固定床反应器中成功建立了水解过程，但需要进一步优化。

引用次数: 0

Antibacterial effects and mechanisms of fisetin-β-cyclodextrin complex under UVA light against Escherichia coli O157:H7 非瑟酮-β-环糊精配合物在UVA光下对大肠杆菌O157:H7的抑菌作用及其机制

IF 3.9 2区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Journal of biotechnology

Pub Date : 2025-12-06 DOI: 10.1016/j.jbiotec.2025.12.003

Tao Liu , Yuzhang Zhu , Jiahui Wang , Kun Lin , Xuan Zhang , Huawei Ma , Chaonan Kong , Wei Cai , Lifang Yang , Mingguo Jiang

The development of natural product photosensitizers was a critical cornerstone for applying photodynamic antimicrobial technology in the food industry. The objective of this study was to fabricate complex of fisetin-β-cyclodextrin (FIT-β-CD), explore their photodynamic antimicrobial efficacy and mechanisms, and characterize it through infrared spectroscopy and X-ray diffraction. The results indicatde that this method can produce good dispersibility and water solubility of FIT-β-CD complex.The photodynamic impact of FIT-β-CD was assessed by examining alterations in reactive oxygen species (ROS) production.The results demonstrated that FIT-β-CD can rapidly generate ROS under UVA, with hydrogen peroxide production being 3.73 times higher than in dark conditions. The antimicrobial efficacy of FIT-β-CD against Escherichia coli O157:H7 (E.coli O157:H7) was evaluated based on bactericidal efficacy, cell membrane damage, DNA and protein damage, motility changes, biofilm formation, and extracellular polysaccharide production. In addition, gene expression analysis by RT-qPCR showed its significant inhibition on the two-component system (TCS) and quorum sensing system (QS). The results indicated that FIT-β-CD exhibited efficient, sensitive photodynamic, and antibacterial properties against E.coli O157:H7, making it a novel photosensitizer. These findings imply potential new applications of fisetin in the field of food antimicrobial agents.

天然产物光敏剂的开发是光动力抗菌技术在食品工业中应用的重要基石。本研究的目的是制备非瑟酮-β-环糊精配合物（FIT-β-CD），探讨其光动力抗菌效果和机制，并通过红外光谱和x射线衍射对其进行表征。结果表明，该方法制备的FIT-β-CD配合物具有良好的分散性和水溶性。通过检测活性氧（ROS）产生的变化来评估FIT-β-CD的光动力学影响。结果表明，在UVA条件下，FIT-β-CD可以快速生成ROS，过氧化氢产量是暗条件下的3.73倍。通过对大肠杆菌O157:H7 （E.coli O157:H7）的杀菌效果、细胞膜损伤、DNA和蛋白质损伤、运动改变、生物膜形成和胞外多糖生成等指标评价FIT-β-CD对大肠杆菌O157:H7的抑菌效果。此外，RT-qPCR基因表达分析显示其对双组分系统（TCS）和群体感应系统（QS）有显著抑制作用。结果表明，FIT-β-CD对大肠杆菌O157:H7具有高效、灵敏的光动力学和抗菌性能，是一种新型光敏剂。这些发现暗示了非瑟酮在食品抗菌剂领域的潜在新应用。

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引用次数: 0

Increasing the chiral purity of cellulosic L-lactic acid by D-Lactate oxidase-catalyzed oxidation in engineered lactic acid bacterium Pediococcus acidilactici d -乳酸氧化酶在工程乳酸菌中催化氧化提高纤维素l -乳酸的手性纯度。

IF 3.9 2区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Journal of biotechnology

Pub Date : 2025-12-05 DOI: 10.1016/j.jbiotec.2025.12.002

Jiao Liu, Chaolong Qu, Bin Zhang, Jie Bao

Trace amounts of D-lactic acid derived from lignocellulose feedstock and nitrogen additives frequently reduce the chiral purity of L-lactic acid below the polymerization standard during biorefinery fermentations. The spontaneous D-lactic acid degradation catalyzed by the highly substrate specific D-lactate oxidase in lactic acid bacterium provides an ideal solution. This study used a D-lactate oxidase GOX2071 with relatively high activity for construction of D-lactic acid oxidation pathway in the L-lactic acid producing strain Pediococcus acidilactici ZY271. The engineered P. acidilactici LJ2071 strain demonstrated an active D-lactic acid oxidation performance while the high L-lactic acid production property was well maintained. A L-lactic acid chiral purity of 99.63 % was obtained using the D-lactic acid containing wheat straw as feedstock and 40 % of corn steep liquor (CSL) as nitrogen additive to alternate expensive yeast extract. This study provided a practical method for upgrading the chirality of cellulosic L-lactic acid using lignocellulose feedstock and cheap nitrogen additives.

在生物精炼发酵过程中，来自木质纤维素原料和氮添加剂的微量d -乳酸经常使l -乳酸的手性纯度低于聚合标准。乳酸菌中具有高度底物特异性的d -乳酸氧化酶对d -乳酸的自发降解提供了理想的解决方案。本研究利用活性较高的d -乳酸氧化酶GOX2071在产l -乳酸的Pediococcus acidacactii ZY271中构建d -乳酸氧化途径。工程菌株LJ2071在保持高产l -乳酸特性的同时，表现出较强的d -乳酸氧化能力。以小麦秸秆为原料，以40%的玉米浸泡液（CSL）为氮添加剂，替代昂贵的酵母浸膏，得到了手性纯度为99.63%的l -乳酸。本研究为利用木质纤维素原料和廉价的氮添加剂提高纤维素l -乳酸的手性提供了一种实用的方法。

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引用次数: 0

Enhancing heterotrophic lutein production in Chlorella protothecoides through combined phytohormone and nitrogen strategies 植物激素和氮素联合策略促进原小球藻异养叶黄素的产生

IF 3.9 2区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Journal of biotechnology

Pub Date : 2025-12-04 DOI: 10.1016/j.jbiotec.2025.12.001

Jiaxin Li , Weiyang Zhao , Yunlei Fu , Ranxuan Li , Jin Liu , Shufang Yang , Jie Zheng , Haijin Mou , Han Sun

Large-scale production of microalgal lutein faces challenges due to limited biomass and lutein content. In this work, we developed an exponential fed-batch model to achieve heterotrophic high-density growth of Chlorella protothecoides CS-41. Then, indole-3-acetic acid (IAA) and high nitrogen were combined to promote lutein accumulation in situ. Fed-batch culture enhanced the TCA cycle of C. protothecoides CS-41 to benefit cell growth while increasing carotenoid flux to facilitate lutein accumulation through redirecting carbon flux towards lutein biosynthesis, improved by IAA. Also, high nitrogen increased lutein content through modulation of CMS (carotenoid methyltransferase synthase), CRTISO (carotenoid isomerase), LCYB (lycopene β-cyclase), ZEP (zeaxanthin epoxidase), and VDE (violaxanthin de-epoxidase). Finally, under heterotrophic conditions, combining 200 mM IAA with 3.6 g/L urea improved pyruvate and 3-phosphoglyceric acid and then boosted lutein content and productivity to 3.27 mg/g and 20.38 mg/L/d, respectively, by increases of 58 % and 56.7 %. Addressing issues including density dilution, container transfer, and in-situ lutein accumulation under heterotrophic high-density conditions offers a cost-effective solution.

由于生物量和叶黄素含量有限，微藻叶黄素的大规模生产面临挑战。本研究建立了小球藻CS-41异养高密度生长的指数补料批量模型。然后，吲哚-3-乙酸（IAA）和高氮配合施用，促进叶黄素原位积累。补料批培养增强了C. protothecoides CS-41的TCA循环，有利于细胞生长，同时增加类胡萝卜素通量，通过将碳通量转向叶黄素的生物合成，促进叶黄素的积累，IAA改善了这一过程。高氮通过调节类胡萝卜素甲基转移酶合成酶（CMS）、类胡萝卜素异构酶（CRTISO）、番茄红素β环化酶（LCYB）、玉米黄质环氧化酶（ZEP）和紫黄质去环氧化酶（VDE）提高叶黄素含量。最后，在异养条件下，200 mM IAA与3.6 g/L尿素配用可改善丙酮酸和3-磷酸甘油酸，叶黄素含量和产量分别提高了3.27 mg/g和20.38 mg/L/d，分别提高了58 %和56.7% %。在异养高密度条件下，解决密度稀释、容器转移和原位叶黄素积累等问题提供了具有成本效益的解决方案。

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引用次数: 0

High specificity of MsmCAR toward 4-hydroxyvaleric acid enables efficient 1,4-pentanediol production from biomass-derived levulinic acid MsmCAR对4-羟戊酸的高特异性使得从生物质衍生的乙酰丙酸中高效生产1,4-戊二醇。

IF 3.9 2区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Journal of biotechnology

Pub Date : 2025-12-01 DOI: 10.1016/j.jbiotec.2025.11.020

Suhye Choi , Seung Hun Lee , Yunyeong Kim , Woo Young Jeon , Jung oh Ahn , Shashi Kant Bhatia , Kwon-Young Choi , Jeong Chan Joo , Yung-Hun Yang

Alkanediols are industrial compounds with diverse applications. However, the biological production of C5 diols is limited by the low substrate specificity and efficiency of carboxylic acid reductases (CARs) toward short to medium-chain substrates. We developed a whole-cell biotransformation system for efficient 1,4-pentanediol (1,4-PDO) production, aimed at developing a platform pathway for diol biosynthesis. By screening 10 CARs, we identified MsmCAR from Mycolicibacterium smegmatis MC2 155 as the most effective for C5 diol biosynthesis, with high specificity toward 4-hydroxyvaleric acid (4-HV). We constructed a whole-cell catalyst expressing MsmCAR and enhanced its performance by introducing ppk2b from Corynebacterium glutamicum ATCC 13032 and chnD from Acinetobacter sp., which encode proteins involved in cofactor regeneration. Systematic optimization of the reaction conditions including buffers, cofactors, metal ions, and cultivation parameters led to a maximum titer of 78.10 mM 1,4-PDO, with a productivity of 1.86 mM/h, when using 4-HV as a substrate. To expand the substrate scope to levulinic acid, 3-hydroxybutyrate dehydrogenase and formate dehydrogenase biosynthetic genes were introduced. This system yielded 22.10 mM 1,4-PDO within 10 h. This work highlights the potential of MsmCAR as a versatile biocatalyst and provides a sustainable strategy for producing short- to medium-chain diols from biomass-derived feedstocks.

烷二醇是一种用途广泛的工业化合物。然而，C5二醇的生物生产受到低底物特异性和羧酸还原酶（CARs）对中短链底物的效率的限制。我们开发了一种高效生产1,4-戊二醇（1,4- pdo）的全细胞生物转化系统，旨在开发二醇生物合成的平台途径。通过筛选10个CARs，我们鉴定出来自垢垢分枝杆菌MC2 155的MsmCAR对C5二醇的生物合成最有效，对4-羟基戊酸（4-HV）具有高特异性。我们构建了表达MsmCAR的全细胞催化剂，并通过引入来自谷氨棒状杆菌ATCC 13032的ppk2b和来自不动杆菌sp.的chnD来增强其性能，这两个基因编码参与辅助因子再生的蛋白质。系统优化反应条件，包括缓冲液、辅因子、金属离子和培养参数，以4-HV为底物时，最大滴度为78.10mM 1,4- pdo，产率为1.86mM/h。为了将底物范围扩大到乙酰丙酸，引入了3-羟基丁酸脱氢酶和甲酸脱氢酶生物合成基因。该体系在10h内生成22.10mM的1,4- pdo。这项工作突出了MsmCAR作为一种多功能生物催化剂的潜力，并为从生物质来源的原料中生产短至中链二醇提供了可持续的策略。

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引用次数: 0

Experimental immobilization of Clostridium histolyticum collagenases on green-synthesized CuO nanoparticles supported by bioinformatics analysis 绿色合成氧化铜纳米颗粒固定化溶组织梭菌胶原酶的实验研究。

IF 3.9 2区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Journal of biotechnology

Pub Date : 2025-11-29 DOI: 10.1016/j.jbiotec.2025.11.019

Farzaneh Barati , Fakhrisadat Hosseini , Parinaz Ghadam , Seyed Shahriar Arab

Enzymes are widely used in various industries due to their eco-friendly nature, high selectivity, and ability to operate under mild conditions. However, large-scale applications are often hindered by challenges such as limited stability, high costs, and difficulties in recyclability. Enzyme immobilization presents a promising strategy to overcome these limitations. Microbial collagenases, particularly those from Clostridium histolyticum, play a crucial role in industrial and biomedical applications. In silico analyses effectively predict enzyme immobilization outcomes, minimizing experimental time and cost. This study employed enzyme surface analysis, a bioinformatics approach, to determine the surface charge type of the support for the physical immobilization of C. histolyticum collagenases. The analysis identified the most suitable regions on the enzyme surfaces for attachment—positively charged and distant from critical sites—ensuring minimal disruption to activity. Copper oxide (CuO) nanoparticles (NPs) were synthesized using walnut green husk extract and subsequently characterized. The green-synthesized CuO NPs were monodisperse and spherical (∼40 nm) with a negatively charged surface, confirming their suitability as supports for enzyme immobilization. Employing the CuO NPs as supports resulted in an immobilization yield of 42.15 % and an activity yield of 146.2 %. Further optimization of immobilization conditions could improve IY. The high AY suggests that immobilization did not hinder enzyme function but may have enhanced enzyme-substrate affinity and increased local substrate concentration, thereby boosting enzyme efficiency.

酶因其生态友好、高选择性和在温和条件下工作的能力而广泛应用于各个行业。然而，大规模应用经常受到诸如稳定性有限、成本高和可回收性困难等挑战的阻碍。酶固定化是克服这些限制的一种很有前途的策略。微生物胶原酶，特别是来自溶组织梭菌的胶原酶，在工业和生物医学应用中起着至关重要的作用。在硅分析有效地预测酶固定结果，最大限度地减少实验时间和成本。本研究采用酶表面分析这一生物信息学方法，确定了溶组织菌胶原酶物理固定载体的表面电荷类型。分析确定了酶表面最适合附着的区域-带正电且远离关键位点-确保对活性的干扰最小。以核桃绿壳提取物为原料合成了氧化铜纳米颗粒，并对其进行了表征。绿色合成的CuO NPs为单分散球形（~40nm），表面带负电荷，证实了其作为固定化酶载体的适宜性。采用CuO NPs作为载体，固定化产率为42.15%，活性产率为146.2%。进一步优化固定条件可以提高IY。高AY表明固定化不会阻碍酶的功能，但可能增强了酶与底物的亲和力，增加了局部底物浓度，从而提高了酶的效率。

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引用次数: 0

Directional lactic acid production could mitigate antibiotic and ARGs pollution 定向乳酸生产可以减轻抗生素和ARGs的污染

IF 3.9 2区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Journal of biotechnology

Pub Date : 2025-11-27 DOI: 10.1016/j.jbiotec.2025.11.014

Fubin Yin , Tianjing Lian , Haoying Qu , Dongan Cui , Xiaomeng Li , Yangyang Cai , Hongmin Dong

The technology of directional lactic acid production (DLAP) for animal manure is an innovative way of value-added utilization, but the influence of residual antibiotics in animal manure on the DLAP is still unclear, especially the degradation rate and degradation pathways of antibiotic with residues lactic acid bacteria (LAB). In this study, the most common residual antibiotic (Sulfachloropyridazine, SCP) and LAB (Lactobacillus bulgaricus, Lactobacillus acidophilus, Lactobacillus plantarum and Lactobacillus casei) were employed as the research objects, the SCP degradation, degradation pathways and antibiotic resistance genes (ARGs) with different LAB were investigated in pure culture. Results showed that LAB has a positive effect on the reduction in ARGs (sul1, sul2, sul3, sulA, intI1 and tnp A). The SCP degradation rates were 90.3 %, 85.1 %, 86.1 % and 84.0 % with the growth of Lactobacillus bulgaricus, Lactobacillus acidophilus, Lactobacillus plantarum and Lactobacillus casei at 50 mg/L, respectively. And the SCP degradation pathway is different for LAB, but the main decomposition functions, including S-N bond cleavage, CH₃ release, SO₂ release, nitration, hydroxylation, NH₂ release and C-N bond cleavage, are similar. And most of intermediate products of SCP were lower toxicity compared to the parent compound. The findings proved the DLAP process as a guidance for a potential solution for reducing antibiotic and ARGs contamination.

畜禽粪便定向产乳酸（DLAP）技术是一种创新的增值利用方式，但畜禽粪便中残留抗生素对DLAP的影响尚不清楚，尤其是抗生素与残留乳酸菌（LAB）的降解速率和降解途径。本研究以最常见的残留抗生素磺胺氯吡嗪（磺胺氯吡嗪）和乳酸菌（保加利亚乳杆菌、嗜酸乳杆菌、植物乳杆菌和干酪乳杆菌）为研究对象，在纯培养中研究了不同乳酸菌对SCP的降解、降解途径和抗生素抗性基因（ARGs）。结果表明，LAB对ARGs （sul1、sul2、sul3、sulA、intI1和tnp a）的减少具有积极作用。当保加利亚乳杆菌、嗜酸乳杆菌、植物乳杆菌和干酪乳杆菌生长在50 mg/L时，SCP的降解率分别为90.3 %、85.1 %、86.1 %和84.0 %。LAB的SCP降解途径不同，但主要分解功能相似，包括S-N键裂解、CH3释放、SO2释放、硝化、羟基化、NH2释放和C-N键裂解。与母体化合物相比，SCP的大部分中间产物毒性较低。研究结果证明，DLAP工艺为减少抗生素和ARGs污染的潜在解决方案提供了指导。

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引用次数: 0

Sodium alginate-immobilized epoxide hydrolase: A multifaceted strategy for enhanced stability, reusability, and catalytic performance 海藻酸钠固定化环氧化物水解酶：提高稳定性、可重用性和催化性能的多方面策略

IF 3.9 2区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Journal of biotechnology

Pub Date : 2025-11-27 DOI: 10.1016/j.jbiotec.2025.11.017

Zongzhong Yu , Meinan Zhu , Xiao Gu , Zhongkun Wu , Peiqin Chen , Chunying Jin , Junning Zhao , Guangya Zhang , Wei Jiang

Immobilized macromolecular enzymes effectively address critical challenges including environmental sensitivity, thermal instability, and non-reusability, thereby establishing the fundamental and practical significance of enzyme immobilization research. This study employed sodium alginate (SA), ZIF-8, and Bi-EA materials to immobilize Aspergillus carlsbadensis-derived epoxide hydrolase (AcEH). Comparative analysis revealed SA's superior immobilization efficiency. Material characterization via SEM, FTIR, and XRD was performed, followed by optimization of SA immobilization parameters. The immobilized enzyme maintained activity over 6 reuse cycles. Incorporating 0.5 wt% carboxymethyl cellulose (CMC) extended AcEH@SA-CMC reusability to 8 cycles. Magnetic modification through Fe₃O₄ powder incorporation yielded AcEH@SA-CMC- Fe₃O₄ hydrogel with enhanced recoverability. Results demonstrate SA's exceptional biocompatibility underpins its immobilization efficacy, while CMC co-entanglement creates a porous network facilitating mass transfer and mechanical reinforcement. Fe₃O₄ powder plays a promoting role in improving enzyme activity. This methodology not only evaluates immobilization matrices but establishes an optimized protocol for macromolecular AcEH immobilization.

固定化大分子酶有效地解决了环境敏感性、热不稳定性和不可重复使用等关键挑战，从而确立了酶固定化研究的基础和现实意义。本研究采用海藻酸钠（SA）、ZIF-8和Bi-EA材料固定化卡尔斯巴登曲霉衍生环氧化物水解酶（AcEH）。对比分析表明，SA具有较好的固定化效果。通过SEM， FTIR和XRD对材料进行了表征，并对SA的固定参数进行了优化。固定化酶在6次重复使用循环中保持活性。加入0.5 wt%羧甲基纤维素（CMC）将AcEH@SA-CMC可重复使用延长至8次循环。通过掺入Fe3O4粉末进行磁改性，得到了回收率提高的AcEH@SA-CMC- Fe3O4水凝胶。结果表明，SA的特殊生物相容性支撑了其固定效果，而CMC的共缠结形成了多孔网络，促进了传质和机械加固。Fe3O4粉末对提高酶活性有促进作用。该方法不仅评价了固定基质，而且建立了大分子AcEH固定的优化方案。

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