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IF 3 3区生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochimie

Pub Date : 2026-01-01 DOI: 10.1016/S0300-9084(25)00306-2

引用次数: 0

ANGPTL4 regulates the adipogenic–osteogenic differentiation balance of bone marrow mesenchymal stem cells: A novel mechanism of osteoporosis from the perspective of lipid metabolism ANGPTL4调节骨髓间充质干细胞成脂-成骨分化平衡：脂质代谢视角下骨质疏松的新机制

IF 3 3区生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochimie

Pub Date : 2025-12-31 DOI: 10.1016/j.biochi.2025.12.014

Fuhao Huang , Zhu Zhu , Qiqiang Xing , Ming Li

To investigate the role of lipid metabolism abnormalities in the progression of osteoporosis (OP), clarify the impact of the key regulator angiopoietin-like protein 4 (ANGPTL4) on the adipogenic–osteogenic differentiation balance of bone marrow mesenchymal stem cells (BMSCs), and provide new insights into the molecular mechanisms and targeted therapy of OP, single-cell and multi-omics transcriptomic datasets were integrated with lipid metabolism–related gene sets. Potential key genes were identified through AUCell scoring, enrichment analysis, and machine learning algorithms validated by 5-fold cross-validation. CellChat was applied to analyze intercellular communication, while GSVA revealed associated signaling pathways. Furthermore, functional validation was performed by knocking down ANGPTL4 in BMSCs using two independent siRNA sequences. The effects on differentiation were assessed by lipid accumulation and osteogenic mineralization assays, biochemical assays, recombinant protein rescue experiments, time-course Western blot, and qPCR analysis of clinical bone marrow samples. Analysis revealed that OP-BMSCs exhibited significantly enhanced lipid metabolism activity. ANGPTL4 was identified as a core candidate gene, demonstrating robust discriminative power with a mean AUC of 0.777 in 5-fold cross-validation. Functional assays confirmed that ANGPTL4 knockdown significantly inhibited adipogenesis while enhancing osteogenic differentiation independent of cell proliferation. Importantly, treatment with recombinant ANGPTL4 protein effectively reversed these phenotypic changes. Mechanistically, ANGPTL4 silencing specifically upregulated BMP2, BMP4, and BMPR1A, leading to the activation of p-Smad1/5/9 and the accelerated expression of Runx2 and Ocn in a time-dependent manner. Consistent with these findings, ANGPTL4 mRNA levels were significantly elevated in bone marrow samples from OP patients. In conclusion, ANGPTL4 serves as a critical checkpoint connecting lipid metabolism and OP pathology. It inhibits osteogenesis by suppressing the BMP2/4-BMPR1A-Smad signaling axis. Targeting ANGPTL4 effectively restores the adipo-osteogenic balance of BMSCs, suggesting it is a promising candidate target for OP therapy, pending further in vivo validation.

为了研究脂质代谢异常在骨质疏松症（OP）进展中的作用，阐明关键调节因子血管生成素样蛋白4 （ANGPTL4）对骨髓间充质干细胞（BMSCs）成脂-成骨分化平衡的影响，并为OP的分子机制和靶向治疗提供新的见解，我们将单细胞和多组学转录组数据集与脂质代谢相关基因集整合在一起。通过AUCell评分、富集分析和5倍交叉验证的机器学习算法确定潜在的关键基因。CellChat应用于分析细胞间通讯，而GSVA揭示了相关的信号通路。此外，通过使用两个独立的siRNA序列敲除BMSCs中的ANGPTL4，进行了功能验证。通过脂质积累和成骨矿化实验、生化实验、重组蛋白救援实验、时间过程Western blot和临床骨髓样本qPCR分析来评估其对分化的影响。分析显示，OP-BMSCs的脂质代谢活性显著增强。ANGPTL4被确定为核心候选基因，5倍交叉验证的平均AUC为0.777，具有较强的判别能力。功能分析证实，ANGPTL4敲低显著抑制脂肪形成，同时增强成骨分化，而不依赖于细胞增殖。重要的是，重组ANGPTL4蛋白治疗有效地逆转了这些表型变化。从机制上讲，ANGPTL4沉默特异性上调BMP2、BMP4和BMPR1A，导致p-Smad1/5/9的激活，并以时间依赖性的方式加速Runx2和Ocn的表达。与这些发现一致，OP患者骨髓样本中ANGPTL4 mRNA水平显著升高。综上所述，ANGPTL4是连接脂质代谢和OP病理的关键检查点。它通过抑制BMP2/4-BMPR1A-Smad信号轴抑制成骨。靶向ANGPTL4有效地恢复了骨髓间充质干细胞的脂肪成骨平衡，这表明它是一个有希望的OP治疗候选靶点，有待进一步的体内验证。

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引用次数: 0

Extracellular mitochondria: a potential player involved in exercise health benefits 细胞外线粒体：参与运动健康益处的潜在参与者。

IF 3 3区生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochimie

Pub Date : 2025-12-26 DOI: 10.1016/j.biochi.2025.12.011

Mafalda Barbosa Pedrosa , Lúcio Lara Santos , Rita Ferreira , José Magalhães

Exercise is widely recognized as an effective nonpharmacological therapy for noncommunicable diseases, with its health benefits mediated in part by exerkines. Recently, extracellular mitochondria (ex-Mito) have been suggested as a player in mediating intercellular communication. While it is known that the health benefits of exercise involve the remodeling of mitochondria in multiple organs, the impact of exercise on circulating ex-Mito is poorly understood. Most existing studies have focused on cell-free circulating mitochondrial DNA, skeletal muscle-derived extracellular vesicles, or platelet-derived mitochondria, without focusing on other types of ex-Mito. The cellular origin of exercise-induced circulating ex-Mito and the role of each form (vesicle-enclosed, free, or as mitochondrial components) in mediating exercise's therapeutic effects are yet to be elucidated. This review aims to delve into the role of ex-Mito as potential players in exercise-related health benefits, paving the way for future research aimed at uncovering the molecular culprits of this nonpharmacological therapy, including mitochondrial transfer and transplantation.

运动被广泛认为是一种有效的非传染性疾病的非药物治疗方法，其健康益处部分是由运动介导的。最近，细胞外线粒体（ex-Mito）被认为是介导细胞间通讯的参与者。虽然我们知道运动对健康的好处涉及到多个器官线粒体的重塑，但运动对循环前水户的影响却知之甚少。大多数现有的研究都集中在无细胞循环线粒体DNA、骨骼肌来源的细胞外囊泡或血小板来源的线粒体上，而没有关注其他类型的前水户线粒体。运动诱导的循环ex-Mito的细胞起源以及每种形式（囊泡封闭、游离或作为线粒体成分）在调节运动治疗效果中的作用尚未阐明。本综述旨在深入研究前水户蛋白在运动相关健康益处中的潜在作用，为未来的研究铺平道路，旨在揭示这种非药物治疗的分子罪魁祸首，包括线粒体转移和移植。

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引用次数: 0

Giardia duodenalis: Base excision repair pathway enzyme FEN1 carries out catalytic activities pertaining to NER pathway 十二指肠贾第虫：碱基切除修复途径酶FEN1具有与NER途径相关的催化活性。

IF 3 3区生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochimie

Pub Date : 2025-12-24 DOI: 10.1016/j.biochi.2025.12.010

Ulises Omar García-Lepe, Sofía Gabriela Tomás-Morales, María Teresa Izaguirre-Hernández, María Luisa Bazán-Tejeda, Rosa María Bermúdez-Cruz

Giardia duodenalis is a binuclear protozoan that causes intestinal infection in humans and animals. The life cycle of G. duodenalis is comprised by 2 stages: trophozoite (vegetative, ploidy: 4 N) and cyst (infective, ploidy: 8–16 N) and the transition from one to another requires a precise coordination as well as the support of the DNA repair machinery. While DNA homologous recombination DNA repair has recently been characterized, NER and BER are pathways that had not been explored. Most of the structure specific enzymes (SSE) participate in a variety of processes like DNA replication stress, DNA adduct repair and Holliday junction processing. In an effort to explore these kinds of enzymes in G. duodenalis, a minimalist parasite, we aimed at characterizing the Fen1 enzyme by cloning its gene to study its catalytic properties (binding and nuclease) using flap and bubble DNA substrates. Unexpectedly, we found that GdFen-1 is able to cleave bubble DNA, then to shed light on which domains of this enzyme are responsible for this activity, giardial acid block and a portion of a cap region were substituted by their human counterparts, and while acid block substitution did not affect this activity, the modification in the cap region did. The possible implications of these findings are addressed.

十二指肠贾第虫是一种双核原生动物，可引起人类和动物肠道感染。十二指肠棘豆的生命周期分为两个阶段：滋养体（营养体，倍性：4N）和囊体（感染体，倍性：8-16N），从一个阶段过渡到另一个阶段需要精确的协调和DNA修复机制的支持。虽然DNA同源重组DNA修复最近已被表征，但NER和BER是尚未探索的途径。大多数结构特异性酶（SSE）参与DNA复制胁迫、DNA加合物修复、Holliday结加工等多种过程。为了在极简寄生虫G. duodenalis中探索这些酶，我们旨在通过克隆Fen1酶基因来研究其催化特性（结合酶和核酸酶），并利用皮瓣和气泡DNA底物研究其催化特性。出乎意料的是，我们发现GdFen-1能够切割气泡DNA，然后阐明该酶的哪个结构域负责这种活性，gi心包酸块和一部分帽区被它们的人类对应物取代，虽然酸块取代不影响这种活性，但帽区修饰会影响这种活性。讨论了这些发现可能产生的影响。

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引用次数: 0

Functional characterization of Cr-CATHs: Novel antimicrobial peptides from the coastal bird Chroicocephalus ridibundus Cr-CATHs的功能表征：来自沿海鸟类的新型抗菌肽。

IF 3 3区生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochimie

Pub Date : 2025-12-18 DOI: 10.1016/j.biochi.2025.12.008

Haoran Dou , Shuangyu Li , Pingchuan Zhang , Zifan Ye , Lili Li , Yipeng Wang , Xudong Jiao

The escalating global threat of antimicrobial resistance (AMR) and chronic biofilm-associated infections underscores the urgent need for novel therapeutic agents. Antimicrobial peptides (AMPs) offer a promising alternative due to their potent activity, broad-spectrum efficacy, and low resistance induction. In this study, we identified three novel cathelicidin-like peptides—Cr-CATH-1 (Cr1), Cr-CATH-2 (Cr2), and Cr-CATH-3 (Cr3)—from Chroicocephalus ridibundus. These peptides were systematically evaluated for their physicochemical properties, antimicrobial activity, bactericidal kinetics, and effects on biofilm formation and persister cells. Cr1 exhibited the most potent and broad-spectrum antimicrobial activity, particularly against Gram-negative and aquatic pathogens. Although Cr2 and Cr3 displayed relatively weaker antimicrobial effects, both peptides were effective in biofilm eradication and persister cell killing. Cr1 also demonstrated strong bactericidal activity, low cytotoxicity, and minimal hemolysis, suggesting a favorable safety profile. Its stability under physiological salt conditions, along with its ability to disrupt bacterial membranes, bind to DNA, and induce reactive oxygen species (ROS) production, highlights its therapeutic potential. In vivo, Cr1 significantly reduced bacterial load and improved survival in a murine peritonitis model, further supporting its potential for clinical and aquaculture applications. Our results suggest that Cr1 is a promising candidate for further development, while Cr2 and Cr3 may provide valuable insights for targeted therapeutic strategies, warranting additional investigation.

抗菌素耐药性（AMR）和慢性生物膜相关感染的全球威胁不断升级，突显了对新型治疗药物的迫切需求。抗菌肽（AMPs）因其有效活性、广谱疗效和低耐药诱导而成为一种有前景的替代方案。在这项研究中，我们从ridibundus中鉴定了三种新的cathelicidin样肽- cr - cath -1 (Cr1), Cr-CATH-2 （Cr2）和Cr-CATH-3 （Cr3）。系统地评估了这些肽的理化性质、抗菌活性、杀菌动力学以及对生物膜形成和持久性细胞的影响。Cr1表现出最有效的广谱抗菌活性，特别是对革兰氏阴性和水生病原体。虽然Cr2和Cr3的抗菌作用相对较弱，但这两种肽都能有效地清除生物膜和杀死持久性细胞。Cr1还显示出很强的杀菌活性、低细胞毒性和最小的溶血，表明其具有良好的安全性。它在生理盐条件下的稳定性，以及它破坏细菌膜、结合DNA和诱导活性氧（ROS）产生的能力，凸显了它的治疗潜力。在体内，Cr1显著降低了小鼠腹膜炎模型中的细菌负荷并提高了存活率，进一步支持了其在临床和水产养殖中的应用潜力。我们的研究结果表明，Cr1是一个有希望进一步开发的候选者，而Cr2和Cr3可能为靶向治疗策略提供有价值的见解，值得进一步研究。

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引用次数: 0

Inhibiting catalytic activity of Plasmodium falciparum aspartate protease plasmepsin V: A biochemical approach to malaria intervention 抑制恶性疟原虫天冬氨酸蛋白酶Plasmepsin V的催化活性：一种生物化学方法干预疟疾。

IF 3 3区生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochimie

Pub Date : 2025-12-13 DOI: 10.1016/j.biochi.2025.12.006

Anitadevi K. Prajapati , Yogesh Kumar , Sinjan Choudhary

Plasmepsin V, an aspartate protease, is crucial for the survival of the malarial parasite Plasmodium falciparum, making it a promising target for antimalarial drug development. In this study, the quinoline-based drugs amodiaquine (AQ) and clioquinol (CQ) are repurposed to inhibit the catalytic activity of PfPlmV. Fluorescence quenching assays suggest that both AQ and CQ exhibit moderate binding affinity for PfPlmV. Temperature-dependent thermodynamic analyses indicate the involvement of both H-bonding/electrostatic interactions and hydrophobic interactions in the binding of AQ/CQ with PfPlmV. Isothermal titration calorimetry (ITC) indicates that these interactions are both enthalpically and entropically favourable. Molecular docking studies show that AQ binds directly to the substrate-binding site of PfPlmV, engaging catalytic dyad residues Asp118 and Asp365, whereas CQ binds to PfPlmV but does not directly interact with the catalytic dyad. Molecular dynamics simulations also corroborated the docking results, revealing that AQ forms a more conformationally stable and hydrophobic interactions driven complex with PfPlmV compared to CQ. Further, the enzyme kinetics studies using fluorogenic substrate demonstrated that AQ efficiently inhibits the catalytic activity of PfPlmV with an IC₅₀ value of (0.42 ± 0.04) μM via competitive inhibition mode. In contrast, CQ fails to bind at the catalytic site and does not exhibit any inhibitory effect on PfPlmV activity. These mechanistic insights will lay the foundations for new biochemical approaches to develop targeted therapies aimed at malaria disease.

Plasmepsin V是一种天冬氨酸蛋白酶，对疟疾寄生虫恶性疟原虫的存活至关重要，使其成为抗疟疾药物开发的一个有希望的靶点。在本研究中，喹啉类药物阿莫地喹（AQ）和氯喹诺（CQ）被用于抑制PfPlmV的催化活性。荧光猝灭实验表明，AQ和CQ对PfPlmV具有中等的结合亲和力。温度相关的热力学分析表明，AQ/ CQ与PfPlmV的结合涉及h键/静电相互作用和疏水相互作用。等温滴定量热法（ITC）表明，这些相互作用在焓和熵上都是有利的。分子对接研究表明，AQ直接与PfPlmV的底物结合位点结合，与催化二偶体Asp118和Asp365残基结合，而CQ与PfPlmV结合，但不直接与催化二偶体相互作用。分子动力学模拟也证实了对接结果，表明与CQ相比，AQ与PfPlmV形成了更稳定的构象和疏水相互作用驱动的复合物。此外，利用荧光底物进行的酶动力学研究表明，AQ通过竞争抑制模式有效抑制PfPlmV的催化活性，IC50值为（0.42±0.04）μM。相比之下，CQ不能在催化位点结合，对PfPlmV活性没有任何抑制作用。这些机制见解将为开发针对疟疾疾病的靶向治疗的新生化方法奠定基础。

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引用次数: 0

Lmod3 ablation disrupts auditory function without compromising hair cell stereocilia development or MET competence Lmod3消融术在不影响毛细胞立体纤毛发育或MET能力的情况下破坏听觉功能。

IF 3 3区生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochimie

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

Haibo Du , Xi Huang , Yixiao Sun , Hao Zhou

F-actin cytoskeleton plays critical roles in hair cell. This study identifies LMOD3, an F-actin nucleator, as specifically expressed in outer hair cells (OHCs) of mice. Lmod3 knockout causes significant hearing loss (about 40 dB elevation in auditory brainstem response (ABR) thresholds, increased distortion product otoacoustic emission (DPOAE) thresholds), while leaving stereocilia morphology and mechanoelectrical transduction (MET) function intact. Phenotypic analyses hint that LMOD3 deficiency likely impairs OHC-based cochlear amplification. Our work establishes LMOD3 as essential for auditory function.

f -肌动蛋白细胞骨架在毛细胞中起重要作用。本研究确定了f -肌动蛋白核子LMOD3在小鼠外毛细胞（ohc）中特异性表达。Lmod3基因敲除会导致严重的听力损失（听觉脑干反应（ABR）阈值升高约40 dB，畸变产物耳声发射（DPOAE）阈值升高），而纤毛立体形态和机电转导（MET）功能完好无损。表型分析提示LMOD3缺陷可能损害ohc为基础的耳蜗放大。我们的研究证实LMOD3对听觉功能至关重要。

引用次数: 0

A comprehensive review on the advanced glycation end products detection: From conventional to advanced approaches 先进糖基化终产物检测的综合综述：从传统方法到先进方法。

IF 3 3区生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochimie

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

Subhrajeet Sahoo , Michael Howsam , Frédéric J. Tessier , Rashmi S. Tupe

The glycation process ultimately generates advanced glycation end products (AGEs) either exogenously or endogenously. With unique structural and functional properties, AGEs contribute to diabetic complications and other glycation-mediated disorders. Owing to the variety of conglomerations, AGEs quantification demands the development of specific and feasible techniques. This article summarizes the molecular mechanism of AGEs formation, along with classifications based on origin, chemical nature, and cellular localization. Moreover, the scope and limitations of existing quantification techniques for both exogenous and endogenous AGEs are discussed. Currently, spectroscopy, chromatography, and immunoassays are widely used methods for measuring AGEs. LC-MS/MS remains the most reliable and specific method, while ELISA and fluorescence spectroscopy offer practical alternatives for specific applications. Non-invasive detection techniques, such as AGEs readers and the facial glycation system, have paved the way; however, associated costs and controversies restrict their feasibility. Hence, future studies with a focus on established and emerging detection techniques for AGEs are crucial to understand the comprehensive role of AGEs in disease progression.

糖基化过程最终产生外源性或内源性的晚期糖基化终产物（AGEs）。由于其独特的结构和功能特性，AGEs与糖尿病并发症和其他糖基化介导的疾病有关。由于聚类的多样性，AGEs的量化需要开发具体可行的技术。本文综述了AGEs形成的分子机制，以及基于来源、化学性质和细胞定位的分类。此外，还讨论了现有外源和内源AGEs定量技术的范围和局限性。目前，光谱法、色谱法和免疫分析法是广泛使用的AGEs测定方法。LC-MS/MS仍然是最可靠和特异性的方法，而ELISA和荧光光谱法为特定应用提供了实用的替代方法。无创检测技术，如AGEs阅读器和面部糖基化系统，已经铺平了道路；然而，相关的成本和争议限制了其可行性。因此，未来的研究将重点放在已建立的和新兴的AGEs检测技术上，这对于了解AGEs在疾病进展中的全面作用至关重要。

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引用次数: 0

Mechanistic insights into NOBA hydrolysis by viper venom secreted phospholipase A2 毒蛇毒液分泌的磷脂酶A2水解NOBA的机理

IF 3 3区生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochimie

Pub Date : 2025-12-05 DOI: 10.1016/j.biochi.2025.12.003

Ana Rita Calixto , Roberto Pinto , Maciej Spiegel , Maria João Ramos , Pedro Alexandrino Fernandes

Snake envenoming remains a major global health challenge, particularly in tropical and subtropical regions. Among the key toxic components of snake venom, secreted phospholipases A₂ (sPLA₂) play a crucial role by hydrolysing cell membrane phospholipids, leading to membrane disruption and severe toxic effects such as inflammation, neurotoxicity, and myotoxicity.

To study sPLA₂ catalytic activity, synthetic soluble substrates like 4-nitro-3-octanoyloxy benzoic acid (NOBA) are widely used in experimental assays, in alternative to membrane phospholipids. However, it is questionable whether mechanistic conclusions taken with small, soluble substrates can be extrapolated to true cell-membrane substrates.

Here, we employed QM/MM calculations to investigate the catalytic mechanism of sPLA₂ using NOBA as a substrate. Our focus was on a sPLA₂ from Bothrops asper. However, the high conservation of sPLA₂ active sites suggests our conclusions should be generalisable to the sPLA₂ of most snake species.

The results reveal a mixed single-water/assisted water mechanism. First, a water molecule, deprotonated by His₄₇, performs a nucleophilic attack on NOBA's carbonyl carbon, with a free energy barrier of 12.8 kcal/mol, resembling the single-water pathway. The collapse of the tetrahedral intermediate and protonation of the leaving group involves two water molecules, resembling the assisted-water pathway. This mixed pathway highlights the catalytic versatility of sPLA₂ and offers new insights into its enzymatic activity with synthetic substrates. Importantly, our finding support NOBA as a valid and suitable substrate for studying the chemical component of sPLA₂ toxicity, even though it does not account for the equally important membrane-binding component of the toxicity mechanism.

蛇中毒仍然是一个重大的全球卫生挑战，特别是在热带和亚热带地区。在蛇毒的关键毒性成分中，分泌磷脂酶A2 （sPLA2）通过水解细胞膜磷脂发挥关键作用，导致细胞膜破坏和严重的毒性作用，如炎症、神经毒性和肌毒性。为了研究sPLA2的催化活性，合成可溶性底物如4-硝基-3-辛烷氧基苯甲酸（NOBA）被广泛用于实验分析，以替代膜磷脂。然而，用小的可溶性底物得出的机械结论是否可以外推到真正的细胞膜底物，这是值得怀疑的。在这里，我们采用QM/MM计算来研究NOBA作为底物的sPLA2的催化机理。我们的重点是Bothrops aspper的sPLA2。然而，sPLA2活性位点的高度保守性表明我们的结论应该适用于大多数蛇种的sPLA2。结果显示单水/助水混合机理。首先，一个被His47去质子化的水分子对NOBA的羰基碳进行亲核攻击，自由能垒为12.8 kcal/mol，类似于单水途径。四面体中间体的坍塌和离去基的质子化涉及两个水分子，类似于辅助-水途径。这种混合途径突出了sPLA2的催化多功能性，并为其与合成底物的酶活性提供了新的见解。重要的是，我们的发现支持NOBA作为研究sPLA2毒性化学成分的有效和合适的底物，即使它不能解释毒性机制中同等重要的膜结合成分。

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引用次数: 0

The CYP134 family in Gram-positive bacteria: From Bacillus to beyond, an orphan P450 lineage awaiting functional discovery 革兰氏阳性细菌中的CYP134家族：从芽孢杆菌到其他，一个等待功能发现的孤儿P450谱系。

IF 3 3区生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochimie

Pub Date : 2025-12-03 DOI: 10.1016/j.biochi.2025.12.001

Yiru Wang , Ying Li

Cytochrome P450 monooxygenases (CYPs) are pivotal enzymes in microbial metabolism, catalyzing a broad spectrum of oxidative reactions with ecological, physiological, and biotechnological significance. Within this superfamily, numerous Bacillus CYPs have been experimentally characterized, yet the CYP134 lineage remains poorly understood and is generally considered an orphan family. Although limited in vitro activities have been observed, a clear physiological substrate or function has not been defined. Despite this uncertainty, CYP134 members are widely distributed across Gram-positive bacteria, particularly within the Firmicutes, and exhibit the conserved structural motifs that safeguard P450 catalytic competence. Their expression is often responsive to environmental cues such as iron limitation and stress, pointing toward potential roles in specialized metabolic circuits and microbial competition. The evolutionary conservation, structural stability, and ecological responsiveness of these enzymes underscore their relevance as emerging candidates for functional discovery. This review integrates current structural, genomic, and ecological perspectives on CYP134 proteins, highlights unresolved questions, and proposes future avenues for exploring their biochemical activities and biotechnological applications.

细胞色素P450单加氧酶（CYPs）是微生物代谢中的关键酶，催化广泛的氧化反应，具有生态，生理和生物技术意义。在这个超家族中，许多CYPs芽孢杆菌已经被实验表征，但CYP134谱系仍然知之甚少，通常被认为是一个孤儿家族。虽然已观察到有限的体外活性，但尚未明确的生理底物或功能。尽管存在这种不确定性，但CYP134成员广泛分布在革兰氏阳性菌中，特别是厚壁菌门中，并表现出保护P450催化能力的保守结构基序。它们的表达通常对环境因素（如铁限制和压力）有反应，指出了在专门的代谢回路和微生物竞争中的潜在作用。这些酶的进化保守性、结构稳定性和生态响应性强调了它们作为功能发现的新兴候选物的相关性。本文综述了目前CYP134蛋白的结构、基因组和生态学观点，强调了尚未解决的问题，并提出了探索其生化活性和生物技术应用的未来途径。

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引用次数: 0