Biochemical Engineering Journal最新文献_第9页

Optimizing Rituximab glycosylation: Insights from single-use bioreactor cultivation 优化利妥昔单抗糖基化：来自一次性生物反应器培养的见解

IF 3.7 3区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Biochemical Engineering Journal

Pub Date : 2025-10-17 DOI: 10.1016/j.bej.2025.109979

Alireza Sattarzadeh , Masoumeh Sadat Mousavi Maleki , Hossein Sedighikamal , Reza Karimi Mostofi , Sadegh Salehi , Meisam Parsianfard

The therapeutic efficacy of monoclonal antibodies (mAbs) is closely related to glycosylation, which affects critical properties, including secretion, solubility, receptor recognition, antigenicity, immunogenicity, bioactivity, and pharmacokinetics. This study evaluated the impact of culture duration and manganese chloride (MnCl₂) concentration on the glycan profile of Rituximab expressed in recombinant Chinese hamster ovary (CHO) cells employing a 200-liter Single-Use Bioreactor (SUB). For this purpose, six experimental groups were designed and conducted with different cultivation times (252, 276, and 300 h) and MnCl2 concentrations (0, 30, 50, and 70 µM). The results demonstrated that cultivation for 276 h with 70 µM MnCl₂ significantly improved galactosylation of the Rituximab glycan profile by 27.24 % relative to the control group without MnCl₂ (P-value <0.05). Additionally, higher levels of fucosylated galactose (G1F & G1F') were observed, likely due to the combined effect of MnCl₂ concentration and optimized culture duration. Peptide mapping by HPLC and SDS-PAGE analyses demonstrated that the primary structure of Rituximab remained unchanged and consistent with MabThera®, with no proteolytic degradation or aggregation.

单克隆抗体（mab）的治疗效果与糖基化密切相关，糖基化影响关键特性，包括分泌、溶解度、受体识别、抗原性、免疫原性、生物活性和药代动力学。本研究采用200升单用途生物反应器（SUB）评估了培养时间和氯化锰（MnCl2）浓度对利妥昔单抗在重组中国仓鼠卵巢（CHO）细胞中表达的聚糖谱的影响。为此，设计了6个不同培养时间（252、276和300 h）和MnCl2浓度（0、30、50和70 µM）的实验组。结果表明，与不添加MnCl 2的对照组相比，添加70 µM MnCl 2培养276 h显著提高了利妥昔单抗聚糖谱的半乳糖基化率27.24 % （p值<；0.05）。此外，观察到较高水平的聚焦半乳糖（G1F & G1F'），可能是由于MnCl 2浓度和优化培养时间的共同作用。HPLC和SDS-PAGE分析显示，利妥昔单抗的初级结构保持不变，与MabThera®一致，没有蛋白水解降解或聚集。

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引用次数: 0

Upgrade of sucrose to glucosyl glycerate and mannitol by a thermostable and minimized three-enzyme cascade 通过耐热性和最小化的三酶级联将蔗糖升级为葡萄糖基甘油和甘露醇

IF 3.7 3区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Biochemical Engineering Journal

Pub Date : 2025-10-17 DOI: 10.1016/j.bej.2025.109983

Zihan Cheng , Juanjuan Liu , Zixuan Huang , Jianyu Wang , Chao Yuan , Dongdong Meng

Upgrading sucrose, the well-established raw material for industrial biomanufacturing, to produce high-value products is of great significance; however, challenges such as its poor atomic economy and low substrate conversion persist in this process. Here, a minimized three-enzyme cascade, featuring theoretically high atomic economy, was designed for the synthesis of value-added glucosyl glycerate (GG) and mannitol from sucrose. Thermophilic sucrose phosphorylase from Bifidobacterium adolescentis, mannitol dehydrogenase from Thermotoga maritima, and formate dehydrogenase from Thiobacillus sp. were purified via a convenient heat treatment and used to construct the enzymatic cascade. Following validation of the feasibility of both the GG synthesis module and the mannitol synthesis module, combined with dual-module integration and enzyme dosage optimization, the three-enzyme cascade was shown to exhibit excellent overall performance. Ultimately, the glucose moiety of sucrose underwent condensation with D-glycerate to yield a high GG concentration of 203.3 mM, achieving a molar substrate conversion of 81.3 % and a productivity of 33.9 mM/h. Concurrently, fructose moiety of sucrose was reduced to 164.5 mM mannitol with a molar substrate conversion of 65.8 %. This thermostable and minimized three-enzyme cascade is characterized by its high atomic economy in converting sucrose into GG and mannitol via green and sustainable enzymatic catalysis. Collectively, this study demonstrates an enzymatic cascade for sucrose upgrading, thereby providing a reference for the value-added utilization of similar substrates.

将蔗糖这一成熟的工业生物制造原料升级为高附加值产品具有重要意义；然而，在这一过程中，其原子经济性差和衬底转化率低等挑战仍然存在。本文设计了一种理论上具有高原子经济性的最小化三酶级联反应，用于从蔗糖合成增值甘油葡萄糖（GG）和甘露醇。分别从双歧杆菌中纯化出嗜热性蔗糖磷酸化酶、从海洋热菌中纯化出甘露醇脱氢酶和从硫杆菌中纯化出甲酸脱氢酶，并将其用于构建酶级联反应。在验证了GG合成模块和甘露醇合成模块的可行性后，结合双模块集成和酶用量优化，三酶级联具有优异的综合性能。最终，蔗糖的葡萄糖部分与d -甘油缩合，得到了高浓度的GG，浓度为203.3 mM，摩尔底物转化率为81.3 %，产率为33.9 mM/h。同时，蔗糖的果糖部分被还原为164.5 mM甘露醇，摩尔底物转化率为65.8% %。这种耐热和最小化的三酶级联反应的特点是通过绿色和可持续的酶催化将蔗糖转化为GG和甘露醇的高原子经济性。综上所述，本研究证明了蔗糖的酶级联升级，从而为类似底物的增值利用提供了参考。

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引用次数: 0

Formate assimilation pathway contributes recombinant protein expression of Komagataella phaffii in bioelectrical carbon dioxide reduction system 甲酸同化途径对法菲Komagataella phaffii在生物电二氧化碳还原系统中的重组蛋白表达有促进作用

IF 3.7 3区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Biochemical Engineering Journal

Pub Date : 2025-10-16 DOI: 10.1016/j.bej.2025.109980

Xiao Zheng , Aibo Feng , Wenjie Cong , Hualan Zhou , Jianguo Zhang

Komagataella phaffii serves as a pivotal industrial microbial cell factory of diverse recombinant proteins and high-value chemicals, which is also developed successfully as a microbial cell using carbon dioxide in the bioelectrical reactor (BER) with results of 1.7-fold recombinant protein production of traditional methanol induction. To elucidate the metabolic shifts of this K. phaffii system, transcriptomic analysis through RNA-seq revealed that 63.8 % of K. phaffii genes were differentially expressed genes (DEGs) significantly, including 1738 up-regulated genes and 1477 down-regulated genes. After gene annotations and enrichment through databases of evolutionary genealogy of genes: non-supervised orthologous groups (EggNOG), gene ontology (GO), and kyoto encyclopedia of genes and genomes (KEGG), of gene analysis involved in methanol metabolic pathway, formate assimilation pathway was found to increase significantly, which was confirmed as the major force of efficient recombinant protein expression by K. phaffii in BER through formaldehyde dehydrogenase gene (FLD) and dihydroxyacetone synthase gene (DAS1) knock out respectively. Therefore, this BER system with engineered K. phaffii open a light window for various efficient recombinant protein expression furtherly from carbon dioxide.

Komagataella phaffii是一个重要的工业微生物细胞工厂，生产多种重组蛋白和高价值化学品，也成功地在生物电反应器（BER）中利用二氧化碳开发了一个微生物细胞，其重组蛋白的产量是传统甲醇诱导的1.7倍。为了阐明该phaffii系统的代谢变化，通过RNA-seq转录组学分析发现63.8 %的phaffii基因显著差异表达基因（deg），其中上调基因1738个，下调基因1477个。通过基因进化谱系数据库进行基因注释和富集后：参与甲醇代谢途径、甲酸代谢途径的基因分析中，发现非监督同源群（EggNOG）、基因本体（GO）和京都基因基因组百科全书（KEGG）显著增加，这证实了K. phaffii通过敲除甲醛脱氢酶基因（FLD）和二羟丙酮合成酶基因（DAS1）在BER中高效表达重组蛋白的主要力量。因此，该系统为进一步利用二氧化碳高效表达多种重组蛋白打开了一扇窗。

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引用次数: 0

Bioremediation of diesel-contaminated saline soil through halotolerant-lipolytic fungal consortium and humic acid 耐盐溶脂真菌联合体和腐植酸对柴油污染盐渍土的生物修复作用

IF 3.7 3区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Biochemical Engineering Journal

Pub Date : 2025-10-15 DOI: 10.1016/j.bej.2025.109969

Ahmed Sarfaraz , Sumbal Sajid , Qingming Zhang , Hengxing Zhang , Lin Lin , Wenfang Chen , Farhan Nabi , Syed Muhammad Mustajab Shah , Obey Kudakwashe Zveushe , Amal Mohamed Omer , Ying Han

Rapid urbanization has resulted in high diesel oil demand and consumption. Such processes are mostly carried out near coastal areas, where accidental oil spills occur during storage and handling, resulting in diesel-contaminated soils. Usually, soil microbial populations can remediate such contamination. However, the remediation of diesel-contaminated saline soils is challenging, as high salt concentrations inhibit the growth and activity of many hydrocarbon-degrading microbes, thereby limiting their remediation potential. In the current study, a combined approach was employed to remediate diesel-contaminated saline soil through bioaugmentation of a halotolerant-lipolytic fungal consortium (HLFC) and humic acid (HA) to alleviate the phytotoxicity of diesel and salt in Lolium perenne in the greenhouse experiment. The highest total petroleum hydrocarbon (TPH) degradation was achieved by the combined application of HLFC-HA (78.1 %), followed by HLFC (74.1 %), HA (65.2 %), and control (CK) (50.8 %). Plants displayed significant stress in diesel-contaminated saline soil. However, the combined application significantly reduced stress, plants showed better growth, photosynthesis, chlorophyll content, and lower antioxidant enzyme activity. Microbial diversity analysis showed that the relative abundances of the fungal genera Scopulariopsis and Aspergillus were significantly high in HLFC and HLFC-HA amended soils. Besides that, soil enzyme activity, including lipase and laccase, significantly increased in HLFC and HLFC-HA-amended soil. These findings demonstrated that HLFC-HA could be a beneficial strategy for the remediation of such oil-contaminated sites.

快速的城市化导致了柴油的高需求和高消耗。这些过程大多在沿海地区附近进行，在储存和处理过程中会发生意外的石油泄漏，导致柴油污染土壤。通常，土壤微生物种群可以修复这种污染。然而，修复柴油污染的盐碱地是具有挑战性的，因为高浓度的盐抑制了许多碳氢化合物降解微生物的生长和活性，从而限制了它们的修复潜力。本研究在温室试验中，采用耐盐溶脂真菌联合体（HLFC）和腐植酸（HA）联合修复柴油污染盐碱地的方法，减轻了柴油和盐对黑麦草（Lolium perenne）的植物毒性。HLFC-HA对总石油烃（TPH）的降解效果最好（78.1 %），其次是HLFC（74.1 %）、HA（65.2% %）和对照（CK）（50.8 %）。在柴油污染的盐渍土壤中，植物表现出明显的胁迫。但配施显著降低了胁迫，植株生长、光合、叶绿素含量均有所提高，抗氧化酶活性降低。微生物多样性分析表明，在HLFC和HLFC- ha改性土壤中，真菌属Scopulariopsis和Aspergillus的相对丰度显著较高。此外，HLFC和HLFC- ha处理的土壤酶活性显著提高，包括脂肪酶和漆酶。这些发现表明，高氟化碳- ha可能是一种有益的石油污染场地修复策略。

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引用次数: 0

Gold nanocages functionalized poly(3-methylthiophene) sensing interface for uric acid assay in clinical serum samples 金纳米笼功能化聚（3-甲基噻吩）传感界面用于临床血清尿酸测定

IF 3.7 3区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Biochemical Engineering Journal

Pub Date : 2025-10-15 DOI: 10.1016/j.bej.2025.109976

Hui Xu , Shengkun Zhang , Zhigang Li , Kexin Lou , Changchun Tu , Baoting Dou , Po Wang , Yujuan Qi

It is crucial to monitor the level of uric acid (UA) in body fluids for the clinical diagnosis and treatment of related diseases. However, the complex composition of human body fluids often means that UA is present alongside substances that have similar chemical structures and redox potentials, making direct determination of UA in biological samples difficult. This study addresses this challenge by modifying the sensing electrode with poly(3-methylthiophene) (P3MT) and assembling gold nanocages (GNCs) to develop an efficient electrochemical sensor for the determination of UA. The inherent catalytic and conductive properties of P3MT provide a suitable substrate and microenvironment, enhancing the assembly efficiency of GNCs. The synergistic catalysis of P3MT and GNCs contributes to the efficient distinguishing of the electrochemical signals of UA against interfering signals. The electrooxidation reaction of UA is found to involve equal proton and electron transfer, and the UA concentration and current display an excellent linear relationship, ranging from 1 to 26 µM under the coexistence of 20 µM ascorbic acid. A detection limit of 0.3 µM is achieved, enabling the sensitive detection of UA in serum samples from both healthy volunteers and gout patients. With its high accuracy, good selectivity, and sensitivity, the sensor is a valuable asset in clinical diagnosis and drug research related to purine metabolism disorders.

监测体液中尿酸水平对临床相关疾病的诊断和治疗具有重要意义。然而，人体体液的复杂组成通常意味着UA与具有相似化学结构和氧化还原电位的物质一起存在，这使得直接测定生物样品中的UA变得困难。本研究通过用聚（3-甲基噻吩）（P3MT）修饰传感电极和组装金纳米笼（GNCs）来开发一种高效的测定UA的电化学传感器，从而解决了这一挑战。P3MT固有的催化和导电特性为GNCs的组装提供了合适的衬底和微环境，提高了GNCs的组装效率。P3MT和GNCs的协同催化作用有助于有效区分UA的电化学信号和干扰信号。在20 µM的抗坏血酸共存条件下，UA的浓度与电流呈良好的线性关系，范围为1 ~ 26 µM。检测限为0.3 µM，可以灵敏地检测健康志愿者和痛风患者血清样品中的UA。该传感器准确度高、选择性好、灵敏度高，在嘌呤代谢紊乱的临床诊断和药物研究中具有重要价值。

{"title":"Gold nanocages functionalized poly(3-methylthiophene) sensing interface for uric acid assay in clinical serum samples","authors":"Hui Xu , Shengkun Zhang , Zhigang Li , Kexin Lou , Changchun Tu , Baoting Dou , Po Wang , Yujuan Qi","doi":"10.1016/j.bej.2025.109976","DOIUrl":"10.1016/j.bej.2025.109976","url":null,"abstract":"<div><div>It is crucial to monitor the level of uric acid (UA) in body fluids for the clinical diagnosis and treatment of related diseases. However, the complex composition of human body fluids often means that UA is present alongside substances that have similar chemical structures and redox potentials, making direct determination of UA in biological samples difficult. This study addresses this challenge by modifying the sensing electrode with poly(3-methylthiophene) (P3MT) and assembling gold nanocages (GNCs) to develop an efficient electrochemical sensor for the determination of UA. The inherent catalytic and conductive properties of P3MT provide a suitable substrate and microenvironment, enhancing the assembly efficiency of GNCs. The synergistic catalysis of P3MT and GNCs contributes to the efficient distinguishing of the electrochemical signals of UA against interfering signals. The electrooxidation reaction of UA is found to involve equal proton and electron transfer, and the UA concentration and current display an excellent linear relationship, ranging from 1 to 26 µM under the coexistence of 20 µM ascorbic acid. A detection limit of 0.3 µM is achieved, enabling the sensitive detection of UA in serum samples from both healthy volunteers and gout patients. With its high accuracy, good selectivity, and sensitivity, the sensor is a valuable asset in clinical diagnosis and drug research related to purine metabolism disorders.</div></div>","PeriodicalId":8766,"journal":{"name":"Biochemical Engineering Journal","volume":"226 ","pages":"Article 109976"},"PeriodicalIF":3.7,"publicationDate":"2025-10-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145359493","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Enhanced biodegradation of n-alkanes in crude oil by immobilized Methylorubrum populi: Enzymatic roles of alkane monooxygenase, cytochrome p450, and lipase 固定化的褐藻增强原油中正构烷烃的生物降解：烷烃单加氧酶、细胞色素p450和脂肪酶的酶促作用

IF 3.7 3区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Biochemical Engineering Journal

Pub Date : 2025-10-15 DOI: 10.1016/j.bej.2025.109977

Mitra Parsa, Sonbol Nazeri

Effective remediation of petroleum contaminants is vital for addressing environmental degradation and advancing sustainable societal development. This study evaluated the hypothesis that immobilization of Methylorubrum populi enhances crude oil biodegradation by improving cell stability and enzymatic activity. Aerobic degradation of 1 % (v/v) crude oil was evaluated over a 7-day period using bacterium strain, Methylorubrum populi under free and immobilized conditions. The analysis included a semi-quantitative assessment of crude oil concentration, revealing that the strain could degrade n-alkanes ranging from C10 to C28. Biodegradation assays revealed that immobilized M. populi achieved a total petroleum hydrocarbon (TPH) degradation of 73 % by day 7, surpassing the 50 % observed in free cells. Analysis of n-alkane degradation (C10–C28) indicated a total degradation ratio of 61 % in immobilized cells versus 45 % in free cells over a 7-day incubation. Moreover, both free and immobilized cell systems could degrade C16–C18 n-alkanes and increased the ratio of C10–C15 fractions on day 3. Enzymatic activity assays of key degradative enzymes—alkane monooxygenase (AlkB), cytochrome P450 (CYP450), and lipase—revealed significantly higher levels in immobilized cells, correlating with enhanced hydrocarbon degradation efficiency. These findings indicate that immobilization improves the robustness and stability of microbial systems, underscoring their potential as practical and eco-friendly tools for petroleum bioremediation.

石油污染物的有效修复对于解决环境退化和促进社会可持续发展至关重要。本研究评估了固定化甲基蓝通过提高细胞稳定性和酶活性来促进原油生物降解的假设。在自由和固定化条件下，研究了菌株Methylorubrum populi在7天内对1 % （v/v）原油的好氧降解效果。分析包括对原油浓度的半定量评估，表明菌株可以降解C10至C28的正构烷烃。生物降解实验表明，固定化的大众分枝杆菌在第7天达到了73 %的总石油烃（TPH）降解，超过了在游离细胞中观察到的50 %。对正烷烃降解（C10-C28）的分析表明，在7天的培养过程中，固定细胞的总降解率为61 %，而游离细胞的总降解率为45 %。此外，游离和固定化细胞体系均能降解C16-C18正构烷烃，并在第3天增加了C10-C15组分的比例。对关键降解酶——烷烃单加氧酶（AlkB）、细胞色素P450 （CYP450）和脂肪酶的酶活性分析显示，固定化细胞中烷烃单加氧酶（AlkB）、细胞色素P450 （CYP450）和脂肪酶的水平显著提高，这与碳氢化合物降解效率的提高有关。这些发现表明，固定化提高了微生物系统的稳健性和稳定性，强调了它们作为石油生物修复的实用和环保工具的潜力。

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引用次数: 0

A high throughput assay to detect enzymatic polyethylene oxidation 一种检测酶促聚乙烯氧化的高通量测定方法

IF 3.7 3区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Biochemical Engineering Journal

Pub Date : 2025-10-15 DOI: 10.1016/j.bej.2025.109978

Ross R. Klauer , Mekhi Williams , Darien K. Nguyen , Megan Tarr , Dionisios G. Vlachos , Kevin V. Solomon , Mark A. Blenner

Biological plastics deconstruction and upcycling have emerged as sustainable alternatives to traditional recycling technologies for plastics waste. The discovery and engineering of efficient thermostable poly(ethylene terephthalate) (PET) hydrolases have made biological PET recycling possible at scale; however, enzymes for non-PET plastics, which account for approximately 70 % of all plastics produced, remain largely undiscovered. To accelerate the discovery of such enzymes, we develop a high-throughput screen to detect initial polymer oxidation, specifically that of the C-H bond to an aldehyde. We test 4-hydrazino-7-nitro-2,1,3-benxoxadiozole hydrazine (NBD-H), which reacts with generated aldehydes to form a fluorescent hydrazone on plasma oxidized low-density polyethylene (LDPE) films. Hydrazone generation correlated well with the area of aldehyde peaks as measured by Fourier Transform Infrared Spectroscopy (FTIR) (R² = 0.92). Moreover, we demonstrate that the probe reliably identifies LDPE-active dye decolorizing peroxidases (DyPs) that generate aldehydes on LDPE films (1.7 – 3.0 fold change relative to background), serving as an effective screen as demonstrated by receiver operating characteristic area under the curve of 0.95. This assay offers an LDPE oxidation screening platform that can be readily parallelized and automated for accelerated discovery of enzymes involved in polyolefin deconstruction.

生物塑料解构和升级回收已经成为传统塑料废物回收技术的可持续替代品。高效耐热型聚对苯二甲酸乙酯（PET）水解酶的发现和工程应用，使PET的生物大规模回收成为可能；然而，非pet塑料的酶，约占所有塑料产量的70% ，在很大程度上仍未被发现。为了加速这种酶的发现，我们开发了一种高通量筛选来检测聚合物的初始氧化，特别是C-H键到醛的氧化。我们测试了4-肼-7-硝基-2,1,3-苯并恶二唑肼（NBD-H），它与生成的醛反应在等离子体氧化低密度聚乙烯（LDPE）薄膜上形成荧光腙。傅里叶变换红外光谱（FTIR）测定的腙生成与醛峰面积有良好的相关性（R2 = 0.92）。此外，我们证明探针可靠地识别LDPE活性染料脱色过氧化物酶（DyPs），这些酶在LDPE薄膜上产生醛（相对于背景变化1.7 - 3.0倍），作为一个有效的筛选，在0.95曲线下的接收器工作特征面积证明了这一点。该分析提供了一个LDPE氧化筛选平台，可以很容易地并行化和自动化，以加速发现涉及聚烯烃解构的酶。

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引用次数: 0

In-depth analysis of acetylcholinesterase: Recent advances in structure, function and assays 乙酰胆碱酯酶的深入分析：结构、功能和检测方法的最新进展

IF 3.7 3区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Biochemical Engineering Journal

Pub Date : 2025-10-14 DOI: 10.1016/j.bej.2025.109974

Jiayi Wu , Lan Wang , Shuo Yang , Siying Pei , Tianwei Liu , Qian Zhou , Xiao Huang , Guanwen Gong , Qian Wang , Wei Liu , Qiong Wu

Acetylcholinesterase (AChE) is a pivotal enzyme in cholinergic neurotransmission, featuring a complex structure and diverse biological roles. This review comprehensively examines recent advances in AChE research, focusing on structure–function relationships, detection methodologies, and practical applications. The well-defined active site architecture is discussed, along with emerging insights into non-catalytic functions in apoptosis, cell adhesion, and disease progression. Significant progress in detection technologies is highlighted, including colorimetric, fluorescence, and multi-platform methods, with particular emphasis on nanozyme-based approaches for organophosphorus pesticide screening and multi-platform strategies for sensitive AChE activity monitoring. The development of multi-target (MT) inhibitors and nano-delivery systems is also discussed to enhance therapeutic efficacy and reduce side effects. Despite these advances, challenges persist in real-time dynamic monitoring, clinical translation of MT agents, and standardized biosensing platforms. This review underscores the need for interdisciplinary efforts to integrate structural biology, sensing technologies, and clinical applications, providing a roadmap for future research aimed at developing high-efficacy, low-toxicity MT drugs and integrated bioanalytical solutions.

乙酰胆碱酯酶（Acetylcholinesterase, AChE）是胆碱能神经传递的关键酶，具有复杂的结构和多种生物学作用。本文综述了乙酰胆碱酯酶的最新研究进展，重点介绍了结构-功能关系、检测方法和实际应用。讨论了明确定义的活性位点结构，以及对细胞凋亡、细胞粘附和疾病进展的非催化功能的新见解。报告强调了检测技术的重大进展，包括比色法、荧光法和多平台方法，特别强调了基于纳米酶的有机磷农药筛选方法和敏感乙酰胆碱酯酶活性监测的多平台策略。本文还讨论了多靶点（MT）抑制剂和纳米递送系统的发展，以提高治疗效果和减少副作用。尽管取得了这些进展，但在实时动态监测、MT药物的临床翻译和标准化生物传感平台方面仍然存在挑战。这篇综述强调了将结构生物学、传感技术和临床应用相结合的跨学科努力的必要性，为未来开发高效、低毒的MT药物和综合生物分析解决方案提供了路线图。

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引用次数: 0

Sulfide removal from wastewater via self-purification by porous media: Effect of flow intermittency and various feed sulfide loadings 多孔介质自净去除废水中的硫化物：流动间歇和不同饲料硫化物负荷的影响

IF 3.7 3区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Biochemical Engineering Journal

Pub Date : 2025-10-13 DOI: 10.1016/j.bej.2025.109975

Lord Richard B. Verania , Hiroyasu Satoh , Tiffany Joan Sotelo

The objective of this study is to evaluate the sulfide removal capacity of a reactor incorporating porous media intermittently exposed to wastewater flow, focusing on the effect of flow intermittency and various feed compositions. To do so, synthetic feed of different initial sulfide concentrations and organic chemical oxygen demand (COD) make-up were introduced to the reactor with different flow frequencies. Changes in oxygen consumption rates and different sulfur species concentrations in the porous media were observed, where oxygen consumption was used as an indicator of COD removal. It was found that sulfide was removed and did not reform in the media within 6 h without flow and at 80 mgS L⁻¹ initial sulfide concentration. For lower initial sulfide concentrations, sulfide was removed within 1 h without flow. The same trends were observed with or without the addition of organic COD to the feed. The results also highlight that flow frequency plays a key role in sulfide oxidation, where more frequent flow yields better performance depending on the sulfide concentration in the feed. Finally, the sulfide removal rates were given on a per porous media volume and area basis. The per volume removal rate was 0.30–0.45 kgS m⁻³ d⁻¹ and the per area removal rate was 1.1–1.9 gS m⁻² d⁻¹.

本研究的目的是评估含多孔介质间歇暴露于污水流的反应器的硫化物去除能力，重点关注流动间歇和不同饲料成分的影响。为此，以不同的流动频率向反应器中引入不同初始硫化物浓度和有机化学需氧量（COD）组成的合成进料。观察了多孔介质中氧消耗速率和不同硫种浓度的变化，其中氧消耗作为COD去除的指标。结果表明，在初始硫化物浓度为80 mg L−1且无流动的情况下，在6 h内硫化物被去除且不发生重整。对于较低的初始硫化物浓度，硫化物在1 h内无流动去除。在饲料中添加有机COD或不添加有机COD时，观察到相同的趋势。研究结果还强调，流动频率在硫化物氧化中起着关键作用，根据进料中的硫化物浓度，流动频率越高，性能越好。最后，给出了每个多孔介质体积和面积的硫化物去除率。单位体积去除率为0.30 ~ 0.45 kgS m−3 d−1，单位面积去除率为1.1 ~ 1.9 gS m−2 d−1。

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引用次数: 0

Kinetic analysis of strictly NADPH-dependent xylose reductase from Debaryomyces nepalensis NCYC 3413 尼泊尔Debaryomyces nepalensis NCYC 3413严格依赖nadph的木糖还原酶动力学分析

IF 3.7 3区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Biochemical Engineering Journal

Pub Date : 2025-10-11 DOI: 10.1016/j.bej.2025.109973

Akilandaeswari J , Benson Rayan , Lawanya Natarajan , Sathyanarayana N. Gummadi

Efficient microbial xylitol production relies on xylose reductase (XR) activity, particularly from robust strains like Debaryomyces nepalensis NCYC 3413. However, the kinetic insights into strictly NADPH-dependent XRs remain scarce. Our study comprehensively characterises the catalytic mechanism of DnXR and the rate-limiting steps through steady-state and transient kinetic analyses. Steady-state kinetics revealed a compulsory ordered sequential bi-bi mechanism, with NADPH binding first. Transient stopped-flow experiments showed that NADPH binding involves two steps, linked to conformational changes in loop 7, while NADP⁺ binding occurs in a single step. Xylose and xylitol exhibited single-step binding to the XR-cosubstrate complex. Multiple-turnover experiments yielded limiting rate constants of 6 ± 0.5 s⁻¹ (reduction) and 1.2 ± 0.3 s⁻¹ (oxidation), without a burst phase. Fluorescence titration confirmed a threefold higher affinity of DnXR for NADPH (

K_{d}

= 1.8 ± 0.3 µM), favouring reduction under physiological conditions. The study provides the first transient kinetic analysis of a strictly NADPH-dependent XR, highlighting the ordered unidirectional reduction of DnXR and its kinetic efficiency for industrial applications.

有效的微生物木糖醇生产依赖于木糖还原酶（XR）活性，特别是来自尼泊尔Debaryomyces NCYC 3413等强大菌株。然而，对严格依赖nadph的xr的动力学见解仍然很少。我们的研究通过稳态和瞬态动力学分析全面表征了DnXR的催化机理和限速步骤。稳态动力学揭示了一个强制有序的顺序bi-bi机制，首先与NADPH结合。瞬时停流实验表明，NADPH结合涉及两个步骤，与环7的构象变化有关，而NADP⁺的结合只发生一个步骤。木糖和木糖醇与xr -共底物的结合为单步结合。多次循环实验得到的极限速率常数为6 ± 0.5 s⁻¹ （还原）和1.2 ± 0.3 s⁻¹ （氧化），没有爆发阶段。荧光滴定证实DnXR对NADPH的亲和力提高了3倍（Kd = 1.8±0.3µM），有利于生理条件下的还原。该研究首次提供了严格依赖nadph的XR的瞬态动力学分析，突出了DnXR的有序单向还原及其工业应用的动力学效率。

{"title":"Kinetic analysis of strictly NADPH-dependent xylose reductase from Debaryomyces nepalensis NCYC 3413","authors":"Akilandaeswari J , Benson Rayan , Lawanya Natarajan , Sathyanarayana N. Gummadi","doi":"10.1016/j.bej.2025.109973","DOIUrl":"10.1016/j.bej.2025.109973","url":null,"abstract":"<div><div>Efficient microbial xylitol production relies on xylose reductase (XR) activity, particularly from robust strains like <em>Debaryomyces nepalensis</em> NCYC 3413. However, the kinetic insights into strictly NADPH-dependent XRs remain scarce. Our study comprehensively characterises the catalytic mechanism of <em>Dn</em>XR and the rate-limiting steps through steady-state and transient kinetic analyses. Steady-state kinetics revealed a compulsory ordered sequential bi-bi mechanism, with NADPH binding first. Transient stopped-flow experiments showed that NADPH binding involves two steps, linked to conformational changes in loop 7, while NADP⁺ binding occurs in a single step. Xylose and xylitol exhibited single-step binding to the XR-cosubstrate complex. Multiple-turnover experiments yielded limiting rate constants of 6 ± 0.5 s⁻¹ (reduction) and 1.2 ± 0.3 s⁻¹ (oxidation), without a burst phase. Fluorescence titration confirmed a threefold higher affinity of <em>Dn</em>XR for NADPH (<span><math><msub><mrow><mi>K</mi></mrow><mrow><mi>d</mi></mrow></msub></math></span> = 1.8 ± 0.3 µM), favouring reduction under physiological conditions. The study provides the first transient kinetic analysis of a strictly NADPH-dependent XR, highlighting the ordered unidirectional reduction of <em>Dn</em>XR and its kinetic efficiency for industrial applications.</div></div>","PeriodicalId":8766,"journal":{"name":"Biochemical Engineering Journal","volume":"226 ","pages":"Article 109973"},"PeriodicalIF":3.7,"publicationDate":"2025-10-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145325507","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

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