Applied Biochemistry and Biotechnology最新文献

Cytotoxic and Cellular Response of Doped Nb-NTO Nanoparticles Functionalized with Mentha arvensis and Mucuna pruriens Extracts on MDA-MB-231 Breast Cancer Cells.

IF 3.1 4区生物学

Applied Biochemistry and Biotechnology Pub Date : 2025-04-03 DOI: 10.1007/s12010-025-05227-y

Muhammad Awais Farooqi, Ji-Hyang Kim, Sehui Kim, Kyeoung Cheol Kim, Hafiz Muhammad Umer Farooqi, Dong-Sun Lee, Chul Ung Kang

{"title":"Cytotoxic and Cellular Response of Doped Nb-NTO Nanoparticles Functionalized with Mentha arvensis and Mucuna pruriens Extracts on MDA-MB-231 Breast Cancer Cells.","authors":"Muhammad Awais Farooqi, Ji-Hyang Kim, Sehui Kim, Kyeoung Cheol Kim, Hafiz Muhammad Umer Farooqi, Dong-Sun Lee, Chul Ung Kang","doi":"10.1007/s12010-025-05227-y","DOIUrl":"https://doi.org/10.1007/s12010-025-05227-y","url":null,"abstract":"Triple-negative breast cancer (TNBC) poses significant challenges as it lacks specific treatment approaches. In this study, we synthesized niobium-nitrogen-doped titanium dioxide (Nb-NTO) nanoparticles (NPs) functionalized with Mentha arvensis ethanolic and Mucuna pruriens methanolic extracts and evaluated their anti-cancer potential against MDA-MB-231 TNBC cells. The functionalization of doped Nb-NTO NPs with Mentha arvensis and Mucuna pruriens extract exhibited significant synergistic effects, reducing cell viability in a dose-dependent manner with enhanced cytotoxicity at lower concentrations compared to individual treatments. Microscopic analysis revealed morphological changes indicative of apoptosis and necrosis, while flow cytometry demonstrated increased apoptotic and necrotic cell populations in the combination-treated groups. These treatments also significantly reduced the secretion of pro-inflammatory cytokines IL-6 and IL-8, suggesting modulation of the inflammatory tumor microenvironment. Furthermore, the functionalized Nb-NTO NPs effectively targeted cancer stem cell (CSC) properties, inhibiting mammosphere formation and clonogenic survival and downregulating critical CSC markers, including c-Myc, OCT4, and NANOG. This study highlights the potential of Nb-NTO NPs functionalized with Mentha arvensis and Mucuna pruriens as a novel therapeutic strategy for TNBC, addressing key hallmarks of cancer, including apoptosis, inflammation, and CSC targeting. While these findings demonstrate promising in vitro anti-cancer efficacy, further in vivo validation and mechanistic studies are necessary to advance these treatments toward clinical anti-cancer applications.","PeriodicalId":465,"journal":{"name":"Applied Biochemistry and Biotechnology","volume":" ","pages":""},"PeriodicalIF":3.1,"publicationDate":"2025-04-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143770826","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"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

The Cyanobacteria Genus Aphanothece: Bioactive Compounds and Applications in Biotechnology.

IF 3.1 4区生物学

Applied Biochemistry and Biotechnology Pub Date : 2025-04-03 DOI: 10.1007/s12010-025-05221-4

Ronald Tarazona Delgado, Rui Dos Santos Ferreira Filho, Carlos Rafael Borges Mendes

{"title":"The Cyanobacteria Genus Aphanothece: Bioactive Compounds and Applications in Biotechnology.","authors":"Ronald Tarazona Delgado, Rui Dos Santos Ferreira Filho, Carlos Rafael Borges Mendes","doi":"10.1007/s12010-025-05221-4","DOIUrl":"https://doi.org/10.1007/s12010-025-05221-4","url":null,"abstract":"Aphanothece is a genus of colonial cyanobacteria with a global distribution that is found in various aquatic and terrestrial environments. It has garnered interest because of its high content of amino acids, carbohydrates, fatty acids, and pigments, which possess bioactive and biotechnological properties. This review analyzes articles highlighting Aphanothece species in biotechnological contexts and describes their biochemical composition. Among its primary metabolites are glutamic acid, alanine, palmitic acid, chlorophyll a, echinenone, and β-carotene. The biotechnological potential of Aphanothece spans the fields of biofuel, health, agro-industry, and bioremediation. The notable bioactivities of species such us A. sacrum, A. pallida, and A. bullosa include photoprotective, immunostimulant, antimicrobial, anticancer, and biostimulant activities due to secondary metabolites such as mycosporine-like amino acids, peptides, betaines, and glycerophospholipids. The high production of hydrogen and lipids by A. halophytica supports its use in biofuels. Species such as A. microscopica are effective at treating agro-industrial and domestic effluents and water polluted by metals and hydrocarbons, alongside simultaneous CO2 capture. This review provides information that can guide the sustainable use of Aphanothece species and identifies gaps in current knowledge, particularly in the development of commercial products. Continuous exploration of this genus can significantly promote environmental sustainability and biotechnological innovation.","PeriodicalId":465,"journal":{"name":"Applied Biochemistry and Biotechnology","volume":" ","pages":""},"PeriodicalIF":3.1,"publicationDate":"2025-04-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143770828","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"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

USP14-Dependent IGF1R Aggravates High Glucose-Induced Diabetic Retinopathy by Upregulating BAP1.

IF 3.1 4区生物学

Applied Biochemistry and Biotechnology Pub Date : 2025-03-31 DOI: 10.1007/s12010-025-05215-2

Li Yu, Xia Zheng, Yan Wu, Kui Ge

{"title":"USP14-Dependent IGF1R Aggravates High Glucose-Induced Diabetic Retinopathy by Upregulating BAP1.","authors":"Li Yu, Xia Zheng, Yan Wu, Kui Ge","doi":"10.1007/s12010-025-05215-2","DOIUrl":"https://doi.org/10.1007/s12010-025-05215-2","url":null,"abstract":"Diabetic retinopathy (DR) is a microvascular complication of diabetes. Insulin-like growth factor 1 receptor (IGF1R) has been implicated in the pathogenesis of DR; however, the underlying mechanism remains unclear. Quantitative reverse transcription polymerase chain reaction (qRT-PCR) was used to assess IGF1R mRNA expression. Western blotting assays were performed to analyze the protein expression of IGF1R, ubiquitin-specific peptidase 14 (USP14), and BRCA1-associated protein 1 (BAP1). Cell viability, apoptosis, interleukin-1 beta (IL-1β), and tumor necrosis factor-alpha (TNF-α) levels were analyzed using cell counting kit-8 assay, flow cytometry, and enzyme-linked immunosorbent assays, respectively. Fluorescent microscopy and flow cytometry were performed for reactive oxygen species (ROS) level assessment, and colorimetric assays for iron (Fe2+) and glutathione (GSH) levels. Co-immunoprecipitation assays and/or colocalization techniques were employed to validate the association of IGF1R with USP14 and BAP1. Treatment with high glucose (HG) increased the protein expression of IGF1R, USP14, and BAP1 in ARPE-19 cells. Silencing of IGF1R mitigated HG-induced apoptosis, inflammatory response, and ferroptosis in ARPE-19 cells. USP14 was found to stabilize IGF1R protein expression through deubiquitination. Overexpression of USP14 exacerbated HG-induced cellular injury, whereas silencing of USP14 protected ARPE-19 cells by reducing IGF1R expression. Interaction between IGF1R and BAP1 was confirmed in ARPE-19 cells and IGF1R silencing protected cells from HG-induced injury by regulating BAP1 expression. Thus, USP14-dependent regulation of IGF1R expression and its interaction with BAP1 play a crucial role in the pathogenesis of high glucose-induced diabetic retinopathy.","PeriodicalId":465,"journal":{"name":"Applied Biochemistry and Biotechnology","volume":" ","pages":""},"PeriodicalIF":3.1,"publicationDate":"2025-03-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143750573","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"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

Lipase Activation by Poly(Methyl Methacrylate) in Dispersed Solution: Mechanistic Insights by Fluorescence Spectroscopy.

IF 3.1 4区生物学

Applied Biochemistry and Biotechnology Pub Date : 2025-03-31 DOI: 10.1007/s12010-025-05217-0

André Merz, Jonas Thelen, Jürgen Linders, Christian Mayer, Kerstin Hoffmann-Jacobsen

{"title":"Lipase Activation by Poly(Methyl Methacrylate) in Dispersed Solution: Mechanistic Insights by Fluorescence Spectroscopy.","authors":"André Merz, Jonas Thelen, Jürgen Linders, Christian Mayer, Kerstin Hoffmann-Jacobsen","doi":"10.1007/s12010-025-05217-0","DOIUrl":"https://doi.org/10.1007/s12010-025-05217-0","url":null,"abstract":"We investigated the mechanisms of polymer-lipase interactions that govern the catalytic activity of lipases in the presence of polymers. Using a combination of fluorescence correlation spectroscopy (FCS), activity analysis, fluorescence spectroscopy, and computational surface analysis, three model lipases-Thermomyces lanuginosus lipase (TLL), Candida antarctica lipase B (CalB), and Bacillus subtilis lipase A (BSLA), with different degrees of hydrophobic active site exposure were studied. Low-molecular-weight poly(methyl methacrylate) (PMMA), synthesized via ARGET ATRP, was employed to study the effect of unstructured polymers in dispersed solution on lipase activity. PMMA significantly enhanced TLL and BSLA hydrolytic activity, while no CalB activation was observed. FCS analysis indicated that this activation was facilitated by polymer lipase binding, a phenomenon observed with TLL and BSLA but not with CalB. Computational analysis further revealed that the surface properties of the lipases were critical for the lipases' susceptibility to activation by PMMA. Although CalB exhibited the largest total hydrophobic surface area, its homogeneous distribution prevented activation, whereas strong, localized hydrophobic interactions allowed PMMA to bind and activate TLL and BSLA. Supported by the quantitative correlation between elevated 8-anilino-1-naphthalenesulfonic acid (ANS) fluorescence in the presence of PMMA and lipase activity, the activation was attributed to locally increased hydrophobicity of the lipases upon polymer binding. These findings provide critical insights into the role of polymer interactions in lipase activation and stabilization, highlighting the potential for designing tailored polymer carriers to optimize enzyme performance in industrial and biotechnological applications.","PeriodicalId":465,"journal":{"name":"Applied Biochemistry and Biotechnology","volume":" ","pages":""},"PeriodicalIF":3.1,"publicationDate":"2025-03-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143750570","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"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

Identification of Enoyl-CoA Hydratase EchA19 in Mycolicibacterium neoaurum Involved in the First β-Oxidation Pathway of Phytosterols for Key Steroidal Intermediate Synthesis.

IF 3.1 4区生物学

Applied Biochemistry and Biotechnology Pub Date : 2025-03-28 DOI: 10.1007/s12010-025-05230-3

Liangyan Zhu, Han Si, Xuemei Li, Na Liu, Jinhui Feng, Qiaqing Wu, Dunming Zhu

{"title":"Identification of Enoyl-CoA Hydratase EchA19 in Mycolicibacterium neoaurum Involved in the First β-Oxidation Pathway of Phytosterols for Key Steroidal Intermediate Synthesis.","authors":"Liangyan Zhu, Han Si, Xuemei Li, Na Liu, Jinhui Feng, Qiaqing Wu, Dunming Zhu","doi":"10.1007/s12010-025-05230-3","DOIUrl":"https://doi.org/10.1007/s12010-025-05230-3","url":null,"abstract":"Microbial transformation has enabled phytosterols as readily available and bio-renewable starting materials for the industrial synthesis of steroidal active pharmaceutical ingredients (APIs). Editing the phytosterol side chain would create various steroidal compounds with a specific C17-side chain, which will greatly facilitate the synthesis of steroidal APIs. Precise cleavage of the phytosterol side chain requires identification of the key enzymes and the reaction pathways of phytosterol side chain metabolism. In this study, a hydratase EchA19 was identified in Mycolicibacterium neoaurum NRRL B-3805, a strain which was engineered by traditional mutation and screening or genetic manipulation, generating recombinant strains for the industrial-scale production of androstenedione (AD), androstadienedione (ADD), and 9α-hydroxy-androstenedione (9α-OH-AD) from phytosterols. It was found that EchA19 is the key hydratase affecting the first β-oxidation pathway of phytosterol side chain metabolism. The previously proposed carboxylation at the C28 position might occur after the cleavage of the C24 branched alkyl side chain, rather than after the dehydrogenation reaction. This study has provided us with new insights and a deeper understanding of the metabolic pathways of phytosterol side chain, and laid a foundation for synthesizing valuable steroid drug intermediates from phytosterols through metabolic regulation by precisely editing the side chain.","PeriodicalId":465,"journal":{"name":"Applied Biochemistry and Biotechnology","volume":" ","pages":""},"PeriodicalIF":3.1,"publicationDate":"2025-03-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143735583","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"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 Tolerance and Growth of Chlorella vulgaris and Scenedesmus quadricauda in Anaerobic Digestate Food Waste Effluent.

IF 3.1 4区生物学

Applied Biochemistry and Biotechnology Pub Date : 2025-03-28 DOI: 10.1007/s12010-025-05220-5

Chunchun Guo, Junhui Chen, Yanfang Ma, Shupeng Lin, Renjie Dong, Roger Ruan, Shan Liu

{"title":"Enhanced Tolerance and Growth of Chlorella vulgaris and Scenedesmus quadricauda in Anaerobic Digestate Food Waste Effluent.","authors":"Chunchun Guo, Junhui Chen, Yanfang Ma, Shupeng Lin, Renjie Dong, Roger Ruan, Shan Liu","doi":"10.1007/s12010-025-05220-5","DOIUrl":"https://doi.org/10.1007/s12010-025-05220-5","url":null,"abstract":"Anaerobic digestate food waste effluent (ADFE) contains high nutrient loads and causes water pollution and waste of resources if discharged without treatment. Microalgae provides a promising strategy for nutrients recovery, biomass production, and CO2 capture. However, due to the characteristics of high ammonia nitrogen and low C/N ratio, it is hard for the original algae to remove N and P in ADFE. Hence, this study investigated the gradient domestication of C. vulgaris and S. quadricauda to enhance their tolerance to high concentrations of ADFE and evaluated their growth and metabolic responses under ADFE concentrations ranging from 20 to 60%. The results revealed that S. quadricauda exhibited better tolerance and growth performance compared to C. vulgaris, with a 42.31% biomass increase in the 40% BG11-diluted ADFE group after domestication. Additionally, extracellular polymeric substances (EPS) secretion increased by 27.66% to help shield algal cells from pollutants. The final removal efficiencies of total nitrogen, total phosphorus, and soluble chemical oxygen demand by S. quadricauda in BG11 dilution group were 37.13%, 40.64%, and 75.45%, respectively, which indicated that domestication alleviated oxidative stress to algal cells. This work demonstrates the simultaneous ADFE treatment and cost-effective microalgal biomass production, providing new insights into microalgal cultivation for the efficient treatment and valorization of nutrient-rich ADFE for sustainability.","PeriodicalId":465,"journal":{"name":"Applied Biochemistry and Biotechnology","volume":" ","pages":""},"PeriodicalIF":3.1,"publicationDate":"2025-03-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143735581","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"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 Thermal Stability/Activity of Geobacillus jurassicus Esterase by Rational Design and Application in the Synthesis of Cinnamyl Acetate.

IF 3.1 4区生物学

Applied Biochemistry and Biotechnology Pub Date : 2025-03-26 DOI: 10.1007/s12010-025-05223-2

Junze Li, Runfei Song, Lin Lin, Tao Li, Yan Yan, Wei Wei, Dongzhi Wei

{"title":"Enhanced Thermal Stability/Activity of Geobacillus jurassicus Esterase by Rational Design and Application in the Synthesis of Cinnamyl Acetate.","authors":"Junze Li, Runfei Song, Lin Lin, Tao Li, Yan Yan, Wei Wei, Dongzhi Wei","doi":"10.1007/s12010-025-05223-2","DOIUrl":"https://doi.org/10.1007/s12010-025-05223-2","url":null,"abstract":"Geobacillus sp. represents an important source of thermophilic esterases, yet studies on the rational design and industrial application of these enzymes remain limited. In our previous research, we identified the esterase Gju768 from Geobacillus jurassicus DSMZ 15726. In the present study, we employed a novel computer-aided rational design approach, ACDP (AutoDock, Consurf, Discovery Studio, PoPMuSiC), to enhance the enzyme's thermal stability. Through molecular docking and conservation analysis, three hotspots were identified. Virtual saturation mutagenesis was subsequently performed, yielding two selected mutations, Q78I and Q78L, from the resulting library. Notably, mutants Q78I and Q78L exhibited significant improvements in thermal stability and enzyme activity compared to the wild type (WT). Compared to WT, mutants Q78I and Q78L exhibited a 65.27% and 38.38% increase in half-life at 65 °C, along with a 14.48% and 1.60% improvement in specific activity at their respective optimal temperatures. Furthermore, under optimized conditions for cinnamyl acetate production, mutant Q78I demonstrated a yield of 68%, compared to only 31% for WT. This study underscored the potential of protein engineering strategies to enhance enzyme performance in industrial applications, particularly for the synthesis of value-added compounds such as cinnamyl acetate.","PeriodicalId":465,"journal":{"name":"Applied Biochemistry and Biotechnology","volume":" ","pages":""},"PeriodicalIF":3.1,"publicationDate":"2025-03-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143727476","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"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

Engineering Escherichia coli for Efficient Production of L-Tryptophan.

IF 3.1 4区生物学

Applied Biochemistry and Biotechnology Pub Date : 2025-03-26 DOI: 10.1007/s12010-025-05228-x

Song Yang, Shengyu Zhou, Quanfeng Liang, Yi Wang, Wei Luo

{"title":"Engineering Escherichia coli for Efficient Production of L-Tryptophan.","authors":"Song Yang, Shengyu Zhou, Quanfeng Liang, Yi Wang, Wei Luo","doi":"10.1007/s12010-025-05228-x","DOIUrl":"https://doi.org/10.1007/s12010-025-05228-x","url":null,"abstract":"L-tryptophan is an essential amino acid widely used in animal feed, food additives, and pharmaceuticals. Although the biosynthesis of L-tryptophan from renewable raw materials is sustainable and environmentally friendly, its long synthetic pathway and complex regulatory mechanisms often result in low production efficiency. In this study, multiple strategies were employed to improve L-tryptophan production in Escherichia coli. These strategies included three main ones: the introduction of anti-feedback inhibition mutations, the overexpression of key rate-limiting enzymes, and the blockage of competitive and catabolic pathways. Additionally, the glucose transport system was modified by replacing it with a non-phosphoenolpyruvate (PEP)-dependent system, thereby reducing PEP consumption. In addition, overexpression of an L-tryptophan transporter protein enhanced L-tryptophan efflux and alleviated product inhibition. Consequently, an efficient L-tryptophan-producing E. coli strain was constructed. Furthermore, dynamic regulation of L-tryptophan synthesis was initially explored by employing a temperature-sensitive promoter to modulate the PEP metabolic pathway. Scale-up fermentation in a 5-L bioreactor demonstrated that the final strain (TY9) produced 34.1 g·L-1 L-tryptophan under fed-batch conditions.","PeriodicalId":465,"journal":{"name":"Applied Biochemistry and Biotechnology","volume":" ","pages":""},"PeriodicalIF":3.1,"publicationDate":"2025-03-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143727475","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"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

Enhancing Chemosensitivity in Drug-Resistant Breast Cancer Cells Using β-Cyclodextrin-Loaded Quercetin and Doxorubicin Inclusion Complex via Modulating SRC/PI3K/Akt Pathway.

IF 3.1 4区生物学

Applied Biochemistry and Biotechnology Pub Date : 2025-03-26 DOI: 10.1007/s12010-025-05219-y

Charan Singh Pawar, Karankumar Balamurugan, Sugumar Baskar, N Rajendra Prasad, Haseeb A Khan

{"title":"Enhancing Chemosensitivity in Drug-Resistant Breast Cancer Cells Using β-Cyclodextrin-Loaded Quercetin and Doxorubicin Inclusion Complex via Modulating SRC/PI3K/Akt Pathway.","authors":"Charan Singh Pawar, Karankumar Balamurugan, Sugumar Baskar, N Rajendra Prasad, Haseeb A Khan","doi":"10.1007/s12010-025-05219-y","DOIUrl":"https://doi.org/10.1007/s12010-025-05219-y","url":null,"abstract":"The challenge of multidrug resistance (MDR) in cancer, which hindered successful chemotherapy, was primarily due to the increased activity of drug efflux transporters in cancer cells. This study explored the potential of a nanosized inclusion complex to overcome drug resistance. The inclusion complex that we prepared contains a beta-cyclodextrin-based formulation encapsulating quercetin (QUE) and doxorubicin (DOX) (β-CD@QD IC), designed to enhance drug delivery and overcome MDR in cancer cells. Through integrative network pharmacology, 92 common targets were identified between QUE and cancer MDR, with SRC kinase emerging as a key target for inhibiting ABCG2 expression in MCF-7/DOX cancer cells. The β-CD@QD IC was formulated via freeze-drying method and characterized using spectroscopic and microscopic techniques. In vitro drug release studies showed that QUE and DOX from β-CD@QD IC exhibited sustained, controlled release, following Higuchi model kinetics. The anticancer efficacy of the complex was tested on MCF-7/DOX cells, where QUE modulated the ABCG2 efflux pump, enhancing the intracellular accumulation of DOX. In vitro assays demonstrated that the inclusion complex significantly increased cell cytotoxicity, induced nuclear condensation, disrupted mitochondrial membrane potential (ΔΨm), elevated reactive oxygen species (ROS) production, and triggered apoptosis-related morphological changes. Hoechst efflux studies demonstrated that QUE effectively inhibited the ABCG2 efflux pump, leading to increased accumulation of Hoechst dye in MCF-7/DOX cancer cells. QUE suppressed SRC kinase signaling, leading to decreased PI3K/Akt expression and reduced ABCG2 overexpression in MCF-7/DOX cells. This study indicated that the β-CD@QD IC loaded with QUE effectively overcame DOX resistance in MCF-7/DOX cells.","PeriodicalId":465,"journal":{"name":"Applied Biochemistry and Biotechnology","volume":" ","pages":""},"PeriodicalIF":3.1,"publicationDate":"2025-03-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143727477","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"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

Hyper-production and Characterization of Exoglucanase Through Physical, Chemical, and Combined Mutagenesis in Indigenous Strain of Thermophilic Aspergillus fumigatus.

IF 3.1 4区生物学

Applied Biochemistry and Biotechnology Pub Date : 2025-03-26 DOI: 10.1007/s12010-025-05222-3

Rabia Ishaq, Muddassar Zafar, Zahid Anwar, Iqra Dildar, Ghazala Mustafa, Tuba Tariq, Mansour Ghorbanpour, Murtaza Hassan

{"title":"Hyper-production and Characterization of Exoglucanase Through Physical, Chemical, and Combined Mutagenesis in Indigenous Strain of Thermophilic Aspergillus fumigatus.","authors":"Rabia Ishaq, Muddassar Zafar, Zahid Anwar, Iqra Dildar, Ghazala Mustafa, Tuba Tariq, Mansour Ghorbanpour, Murtaza Hassan","doi":"10.1007/s12010-025-05222-3","DOIUrl":"https://doi.org/10.1007/s12010-025-05222-3","url":null,"abstract":"The present study explored the optimization of exoglucanase production from waste cellulosic biomaterials using microbial cellulases, focusing on enhancing enzyme efficiency through mutagenesis techniques. Research illustrated the hyper-production and quantitative characterization of an exoglucanase from a thermophilic Aspergillus fumigatus strain via physical and chemical mutagenesis under optimized fermentation conditions. Physical mutagenesis via UV irradiation (15-min exposure) yielded the highest activity (96.57 U/mL), while chemical mutagenesis with ethyl methane sulfonates (250 µg/mL) resulted in 69.61 U/mL activity. Combined mutagenesis using EMS (250 µg/mL) concentration with 15-min UV exposure significantly enhanced exoglucanase production to 136.19 U/mL as compared to the native enzyme 52.46 U/mL. Among various cellulosic substrates, peanut shells exhibited superior suitability for exoglucanase production reaching a maximum activity of 202.41 U/mL. Fermentation parameters including pH, temperature, incubation period, and inoculum size were optimized, leading to a substantial increase in exoglucanase activity of 285.28 U/mL using response surface methodology followed by gel filtration chromatography. The mutant exoglucanase was characterized by its enhanced activities with a higher Vmax (0.6515) and lower Km (0.3142) than those of native enzyme. The characterization has confirmed the temperature and pH tolerance of the mutant enzyme, as well as its tolerance to metal ions and substrate concentrations. This study showed how mutagenesis-driven optimization could provide a means to enhance exoglucanase production from cellulosic biomass, with a rational insight toward enzyme kinetics and applications toward bioenergy generation.","PeriodicalId":465,"journal":{"name":"Applied Biochemistry and Biotechnology","volume":" ","pages":""},"PeriodicalIF":3.1,"publicationDate":"2025-03-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143727039","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"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