Biotechnology and applied biochemistry最新文献

{"title":"Influence of Cell Physiological Status on the Intensified Fed-Batch Cultures at Ultra-High Seeding Density.","authors":"Qingyuan Ran, Ying Su, Weijian Zhang, Xinran Zhang, Liang Zhao, Min Chen, Yuxiang Wan, Wen-Song Tan, Qian Ye","doi":"10.1002/bab.2721","DOIUrl":"https://doi.org/10.1002/bab.2721","url":null,"abstract":"<p><p>Recent years, intensified fed-batch culture with ultra-high seeding density (uHSD-IFB) is coming to the forefront of manufacturers' choice for its enhanced productivity. However, the effects of seed cell physiological state and aeration strategies on these processes remain underexplored due to the ultra-high seeding density. Currently, the pre-production seeding inoculum (N-1) crucial for the uHSD-IFB cultures relies heavily upon case-by-case empirical experiences. To develop a rational seeding approach as a guideline, we here explored the impact of perfusion rates and cell growth states on the subsequent uHSD-IFB processes. It was found that seed cells in the exponential growth phase with high perfusion rates in the N-1 perfusion stage allowed for higher viable cell density and titer in the production stage. In particular, lower levels of reactive oxygen species, higher proportions of G1 and S phase, and higher specific cell oxygen uptake rates (OURs) were exhibited in these cells, resulting in higher cell specific growth rates and integral of viable cell concentration (IVCC) throughout the production cultures. Further investigation into the effect of aeration strategies was carried out in the benchtop bioreactors. A final yield of 4.5 g/L, an increase of nearly 110%, was achieved by a sophisticated dual sparger system compared to the other two processes with either one l-shaped or micro-sparger. These results provide a direction for the design and establishment of high-titer processes in intensified fed-batch cultures at ultra-high seeding density. Synopsis: In this work, we first explored the impact of perfusion rates and cell growth states on the subsequent uHSD-IFB processes. Further investigation into the effect of aeration strategies of intensified fed-batch process was carried out in the benchtop bioreactors. These results provide a direction for the design and establishment of high-titer processes in intensified fed-batch cultures at ultra-high seeding density.</p>","PeriodicalId":9274,"journal":{"name":"Biotechnology and applied biochemistry","volume":" ","pages":""},"PeriodicalIF":3.2,"publicationDate":"2025-01-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143000416","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}

{"title":"Optimization of Culture Medium Ingredients and Culture Conditions for Bacteriocin Production in Lactococcus lactis NCU036019.","authors":"Ziqi Wei, Pengrong Fan, Bo Li, Philippe Madjirebaye, Zhen Peng, Tao Xiong","doi":"10.1002/bab.2714","DOIUrl":"https://doi.org/10.1002/bab.2714","url":null,"abstract":"<p><p>Bacteriocin lactococcin036019 was identified and characterized from Lactococcus lactis NCU036019, which displayed significant antibacterial activity toward foodborne pathogenic bacteria Staphylococcus aureus under various conditions. However, the in situ low-level expression of lactococcin036019 severely limited its wide application in food industry. In this study, we optimized the medium ingredients and culture conditions of L. lactis NCU036019 for maximum production of lactococcin036019. The effects of different carbon sources, nitrogen sources, inorganic salts, growth factors, surfactants, and buffer salts on the production of bacteriocin were studied using antibacterial titer and diameter of inhibitory zone as evaluation indexes. Through single-factor experiments, Plackett-Burman (PB) experiment, steepest ascent experiment and response surface methodology, yeast extract, zinc sulfate, sodium acetate, mannitol, Tween-80, and dipotassium hydrogen phosphate were identified to display significant influence on the production of bacteriocin. By optimizing Man Rogosa and Sharpe (MRS) culture medium ingredients, the antibacterial activity of lactococcin036019 in the cell-free supernatant raised from 46.19 to 300.14 Au/mL, namely, 6.5 times increased. Furthermore, the culture conditions, such as inoculation amount, culture time, and culture temperature, were optimized, and this further increased the antibacterial activity to 409 Au/mL, namely, 8.8 times increased. This study investigated the effects of culture media and conditions on the production of lactococcin036019, and they were optimized for a maximum harvest of bacteriocin, and the significant increase of bacteriocin production in L. lactis NCU036019 facilitates the application of the antibacterial substance in future work.</p>","PeriodicalId":9274,"journal":{"name":"Biotechnology and applied biochemistry","volume":" ","pages":""},"PeriodicalIF":3.2,"publicationDate":"2025-01-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143000418","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}

{"title":"Lipocalin-2 Determination on Multiwalled Carbon Nanotube Integrated Circular Electrodes for Diagnosing Ulcerative Colitis.","authors":"Guangchao Qiao, Jijian Sheng","doi":"10.1002/bab.2715","DOIUrl":"https://doi.org/10.1002/bab.2715","url":null,"abstract":"<p><p>Ulcerative colitis affects the inner lining of the large intestine, causing discomfort, pain, and digestion issues, and sometimes leading to life-threatening complications. With proper medication, symptoms and inflammation can be reduced, improving the condition. In this research, a multiwalled carbon nanotube (MWCN)-modified circular interdigitated electrode (circular-IDE) biosensor was developed to detect the ulcerative colitis biomarker lipocalin-2 and measured at 0-2 V. A dual probing strategy with aptamer and antibody on gold nanoparticles was employed for the detection of lipocalin-2. Probe immobilization was optimized on MWCN-modified circular-IDE, and saturation of 800 nM of aptamer on the GNP-antibody facilitated the identification of lipocalin-2 at concentrations as low as 1 pg/mL, with an R² value of 0.9716 [y = 2.1058x - 2.7351]. Furthermore, lipocalin-2 spiking in serum increased the current responses in correlation with the concentrations of lipocalin-2, indicating selective identification without interference. In addition, nonimmune antibody and GNP-conjugated complementary aptamer did not increase the current responses, affirming the specific detection of lipocalin-2. This MWCN-modified circular-IDE biosensor, utilizing aptamer-antibody interactions, aids in identifying the condition of ulcerative colitis.</p>","PeriodicalId":9274,"journal":{"name":"Biotechnology and applied biochemistry","volume":" ","pages":""},"PeriodicalIF":3.2,"publicationDate":"2025-01-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143000417","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}

{"title":"Assessment of Garbage Enzyme as a Bioremediation Method for the Wastewater Treatment.","authors":"Anoop Narain Singh, Maya Rajnarayan Ray, Umank Mishra, Kruti B Jethwa, Nishant Yadav, Nisha Shankhwar, Gulab Singh Chauhan, Kundan Meshram","doi":"10.1002/bab.2720","DOIUrl":"https://doi.org/10.1002/bab.2720","url":null,"abstract":"<p><p>This study evaluates the efficacy of garbage enzyme (GE) in bioremediation to reduce pollutants in sewage drains that discharge into the natural streams and rivers. Garbage enzyme is prepared with help of brown sugar, fruit, vegetable wastes, and water in the proportion 1:3:10 (by weight), which is then applied to the samples collected from various drainage sites in Jaunpur district, Uttar Pradesh, India. Different concentrations of GE (ranging from 0% to 20%) are mixed with sewage to assess pollution reduction. Different parameters, that is, pH, dissolved oxygen (DO), total suspended solids (TSS), total dissolved solids (TDS), biochemical oxygen demand (BOD), chemical oxygen demand (COD), and CD have been measured on 3rd, 5th, 7th, 10th, 15th, and 20th days. It has been observed in the study that significant percentage reduction in BOD, COD, TSS, and TDS of 80.6%, 62.9%, 74.29%, and 43.9%, respectively, while an increase in DO is 67.8% on 20th day of experiment with 20% GE addition. The result of the study reveals that there is highest reduction occurring on the 20th day with a 20% GE concentration. It has been observed that the GE possesses protease, amylase, and lipase activity. Meaning GE contains microorganisms that breakdown carbohydrates, fats, and proteins and it hydrolyzes these complex organic molecules into simpler compounds, making them more readily biodegradable and that is the reason for substantial reduction in the pollutants. Moreover, in this study it has been observed that there is a notable decrease in foul odor emanating from the samples. Time-dependent fluctuations in pH, TDS, DO, BOD, COD, and temperature are documented, with their correlations examined. This investigation underscores GE's potential in pollution mitigation, particularly in sewage systems, and offers valuable insights for sustainable environmental management practices aimed at conserving natural water bodies. Further tests are required to check the efficiency of treatment with GE dosage higher than 20%.</p>","PeriodicalId":9274,"journal":{"name":"Biotechnology and applied biochemistry","volume":" ","pages":""},"PeriodicalIF":3.2,"publicationDate":"2025-01-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143000415","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}

{"title":"Synthesis and Characterization of Brucine Gold Nanoparticles for Targeted Breast Cancer Therapy: Mechanistic Insights Into Apoptosis and Antioxidant Disruption in MCF-7 Cells.","authors":"Saravanan Alamelu, Kamalesh Balakumar Venkatesan, Manoj Kumar Srinivasan, Pugalendhi Pachaiappan","doi":"10.1002/bab.2719","DOIUrl":"https://doi.org/10.1002/bab.2719","url":null,"abstract":"<p><p>Globally, breast cancer continues to be the leading type of cancer affecting women, with rising mortality rates projected by 2030. This highlights the importance of developing new, affordable treatments, like drug delivery systems that use nanoparticles. Gold nanoparticles (AuNPs), including their exceptional optical and physical attributes, make them an attractive vehicle for targeted treatment, allowing for accurate and focused delivery of medication directly to cancerous cells while reducing harmful side effect. This study focuses on the synthesis and characterization of brucine-gold nanoparticles (BRU-AuNPs) for targeted breast cancer therapy by evaluating their antioxidant and apoptotic mechanism. BRU-AuNPs were synthesized and characterized (UV-Vis spectroscopy, Fourier transform infrared [FTIR], scanning electron microscopy [SEM], x-ray diffraction [XRD], dynamic light scattering [DLS], and zeta potential) to confirm successful synthesis, size, and stability. In vitro studies were assessed using MCF-7 breast cancer cell lines to evaluate cell cytotoxicity, antioxidant balance, reactive oxygen species (ROS) generation, mitochondrial membrane potential, apoptosis induction, cell migration, and pro-apoptotic gene expression. Characterization results confirmed the successful synthesis of BRU-AuNPs with an average crystal size of 85.40 nm and stable surface charge. Results demonstrated that BRU-AuNPs reduced MCF-7 cell viability in a dose-dependent manner, with an IC₅₀ value of 11.47 µg/mL. Treatment with BRU-AuNPs altered the antioxidant balance, increased ROS generation, depolarized mitochondrial membranes, and induced apoptosis. Additionally, BRU-AuNPs inhibited cell migration and upregulated pro-apoptotic gene expression. The synthesized BRU-AuNPs exhibit potential as a highly effective targeted delivery system for breast cancer treatment. Their ability to directly deliver BRU to tumor cells while reducing side effects and enhancing therapeutic efficacy underscores their promise in advancing breast cancer therapy. Further studies are warranted to explore their clinical potential and optimize therapeutic outcomes.</p>","PeriodicalId":9274,"journal":{"name":"Biotechnology and applied biochemistry","volume":" ","pages":""},"PeriodicalIF":3.2,"publicationDate":"2025-01-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142944966","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}

{"title":"Dual Targeting of MEK1 and Akt Kinase Identified SBL-027 as a Promising Lead Candidate to Control Cell Proliferations in Gastric Cancer.","authors":"Maha Alamodi Alghamdi, Hemali Deshpande","doi":"10.1002/bab.2716","DOIUrl":"https://doi.org/10.1002/bab.2716","url":null,"abstract":"<p><p>Dual inhibition of Akt and MEK1 pathways offers a promising strategy to enhance treatment efficacy in gastric cancer. In this study, we employed computational approaches followed by in vitro validations. Our results demonstrate that SBL-027 exhibits robust and enduring interactions with Akt and MEK1 kinases, as evidenced by atomistic molecular dynamics simulations and molecular mechanics Poisson-Boltzmann surface area (MM-PBSA) based binding free energy estimates. The predicted Gibbs binding free energies indicate highly favorable interactions between SBL-027 and both Akt and MEK1 kinases. In vitro, SBL-027 displayed an IC₅₀ value of 195.20 nM against Akt and 239.10 nM against MEK1 enzymes. The compound exhibited potent inhibition of cell proliferation in KATOIII and SNU-5 cells, with GI₅₀ values of 490.70 and 615.14 nM, respectively. Moreover, SBL-027 induced an increase in the sub G₀/G₁ population during the cell cycle of KATOIII and SNU-5 cells, while facilitating early and late-phase apoptosis in these cell lines. Notably, the compound significantly reduced the percentage of dual-positive cells expressing both MEK1 and Akt in gastric cancer cells. The strong binding affinity, stability, and favorable thermodynamics of SBL-027 along with the established in vitro efficacy highlight its potential as a lead compound for further preclinical and clinical development.</p>","PeriodicalId":9274,"journal":{"name":"Biotechnology and applied biochemistry","volume":" ","pages":""},"PeriodicalIF":3.2,"publicationDate":"2025-01-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142944965","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}

{"title":"Harnessing Microbial Signal Transduction Systems in Natural and Synthetic Consortia for Biotechnological Applications.","authors":"Ahmadullah Zahir, Peter A Okorie, Veronica N Nwobasi, Esther I David, Rita O Nwankwegu, Fidelis Azi","doi":"10.1002/bab.2707","DOIUrl":"https://doi.org/10.1002/bab.2707","url":null,"abstract":"<p><p>Signal transduction is crucial for communication and cellular response in microbial communities. Consortia rely on it for effective communication, responding to changing environmental conditions, establishing community structures, and performing collective behaviors. Microbial signal transduction can be through quorum sensing (QS), two-component signal transduction systems, biofilm formation, nutrient sensing, chemotaxis, horizontal gene transfer stress response, and so forth. The consortium uses small signaling molecules in QS to regulate gene expression and coordinate intercellular communication and behaviors. Biofilm formation allows cells to adhere and aggregate, promoting species interactions and environmental stress resistance. Chemotaxis enables directional movement toward or away from chemical gradients, promoting efficient resource utilization and community organization within the consortium. In recent years, synthetic microbial consortia have gained attention for their potential applications in biotechnology and bioremediation. Understanding signal transduction in natural and synthetic microbial consortia is important for gaining insights into community dynamics, evolution, and ecological function. It can provide strategies for biotechnological innovation for enhancing biosensors, biodegradation, bioenergy efficiency, and waste reduction. This review provides compelling insight that will advance our understanding of microbial signal transduction dynamics and its role in orchestrating microbial interactions, which facilitate coordination, cooperation, gene expression, resource allocation, and trigger specific responses that determine community success.</p>","PeriodicalId":9274,"journal":{"name":"Biotechnology and applied biochemistry","volume":" ","pages":""},"PeriodicalIF":3.2,"publicationDate":"2024-12-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142909333","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}

{"title":"Enhanced Synthesis, Purification, and Characterization of a Marine Bacterial Consortium-Derived Protease Enzyme With Destaining and Keratinolytic Activity.","authors":"Rashmi Thakor, Harsh Mistry, Hesham S Almoallim, Mohammad Javed Ansari, Ashish Patel, Virendra Kumar Yadav, Dipak Kumar Sahoo, Himanshu Bariya","doi":"10.1002/bab.2711","DOIUrl":"https://doi.org/10.1002/bab.2711","url":null,"abstract":"<p><p>Two marine-derived bacteria, Bacillus paralicheniformis (HR-1) and Bacillus haynesii (HR-5), were isolated from sediments and identified using 16S ribosomal RNA gene amplification and sequencing as well as biochemical analysis. The development of a bacterial consortium (HR-1 & HR-5) from these two bacteria was used to increase the production of the protease enzyme under various conditions, including fermentation media, carbon and nitrogen sources (1% w/v), different pH levels, incubation time, and the obtained enzyme, were detected using SDS-PAGE followed by purification. Bacterial consortium HR-1 & HR-5 exhibited maximum protease production (330.42 ± 4.47 U/mL) than the individual isolates HR-1 (156.32 ± 2.14 U/mL) and HR-5 (185.73 ± 5.14 U/mL) on supplementing peptone (1% w/v), 2.8% skim milk + N-broth, pH 9, and dextrose (1% w/v) after 48 h of incubation time. The purified enzyme showed increased activity at alkaline pH 9.0 and also in the presence of ions such as Ca⁺², Fe⁺³, Mg⁺², and Mn⁺². The purified protease obtained from the consortium HR-1 and HR-5 shows improved efficiency for stain removal from cloth as well as high keratinolytic efficiency for poultry feather degradation, making this enzyme suitable for industrial use, particularly in the textile and tannery sectors.</p>","PeriodicalId":9274,"journal":{"name":"Biotechnology and applied biochemistry","volume":" ","pages":""},"PeriodicalIF":3.2,"publicationDate":"2024-12-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142909330","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}

{"title":"PRKAA2 Promotes Tumor Growth and Inhibits Ferroptosis through SLC7A11/GSH/GPX4 Pathway in Non-Small Cell Lung Cancer.","authors":"Zhiqiang Wei, Zhilian Zhou, Yu Zhang, Jie Wang, Ke Huang, Yuanyu Ding, Yingming Sun, Mingming Gu, Xiangang Kong, Erping Xi, Shaoshan Zeng","doi":"10.1002/bab.2710","DOIUrl":"https://doi.org/10.1002/bab.2710","url":null,"abstract":"<p><p>Non-small cell lung cancer (NSCLC) is the most pervasive sort of lung cancer with deadly outcome. According to recent studies, a number of neoplastic disorders and ferroptosis are intimately connected. This study aims to identify the role of key ferroptosis-related gene (protein kinase AMP-activated catalytic subunit alpha 2, PRKAA2) and explore new directions for the diagnosis and treatment of NSCLC. The PRKAA2 expression and its influence on survival were analyzed in multiple public databases (TCGA, TIMER2.0, and GEPIA). And PRKAA2 mRNA level in NSCLC cells were examined by qRT-PCR. Silencing of PRKAA2 (sh-PRKAA2) were used to cell transfection. CCK-8, EdU, and flow cytometry assays were used to measure cell proliferation and apoptosis. The protein levels of ferroptosis markers (SLC7A11, GPX4, and NRF2) were determined by western blotting. Meanwhile, the related ferroptosis analysis, such as malondialdehyde (MDA) and glutathione (GSH), reactive oxygen species (ROS), iron, and Fe²⁺ levels were also detected in the transfected cells. Moreover, the relationship between PRKAA2 expression and SLC7A11 was analyzed. NSCLC xenograft mouse models were used for in vivo verification of the PRKAA2 function. Here, our data revealed that PRKAA2 was upregulated in NSCLC cells. Additionally, PRKAA2 strengthened cell proliferation and attenuated apoptosis and ferroptosis of NSCLC cells. The depletion of PRKAA2 enhanced the erastin-induced inhibition effect on cell growth, and notably increased the levels of MDA, ROS, iron, and Fe²⁺, while decreased GSH level in NSCLC cells. In the mechanism exploration, we discovered that PRKAA2 could activate the SLC7A11/GSH/GPx4 antioxidant pathway. The rescue experiments showed that SLC7A11 abrogated the inhibitive impacts of PRKAA2 repression on cellular proliferation, cell apoptosis, and ferroptosis in NSCLC. Besides, animal experiments proved that PRKAA2 enhanced NSCLC tumor growth in vivo. The results discovered that PRKAA2 accelerated the malignant progression, diminished apoptosis and ferroptosis in NSCLC through SLC7A11/GSH/GPX4 pathway. This study provide a novel target in the application of PRKAA2 for NSCLC treatment.</p>","PeriodicalId":9274,"journal":{"name":"Biotechnology and applied biochemistry","volume":" ","pages":""},"PeriodicalIF":3.2,"publicationDate":"2024-12-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142892433","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}

{"title":"Biochemical and In Silico Aspects of Active Compounds From Nyctanthes arbor-tristis Flower As Antidiabetic Agent.","authors":"Saleh ALNadhari, Waleed A A Alsakkaf, Faisal Abdulaziz Albarakat","doi":"10.1002/bab.2709","DOIUrl":"https://doi.org/10.1002/bab.2709","url":null,"abstract":"<p><p>Targeting alpha-glucosidase (maltase-glucoamylase [MGAM] and sucrase-isomaltase [SI]) under diabetes conditions is important to overcome hyperglycemia. Moreover, it is necessary to mitigate hyperglycemia-mediated oxidative stress to evade the progression of diabetes-associated secondary complications. Hence, in the present study, under-explored Nyctanthes arbor-tristis flowers (NAFs) were studied for inhibition of alpha-glucosidase activities. The NAF methanolic extract (NAFME) was prepared. Through liquid chromatography/electrospray ionization tandem mass spectrometry (LC-ESI/MS/MS) analysis, various phytocompounds belonging to different classes-flavonoids, iridoid glycosides, proanthocyanidin, anthocyanin, polyphenol, phenolic acid, fatty acid ester, and carotenoid-were identified. NAFME showed in vitro antioxidant activity. NAFME inhibited maltase, sucrase, glucoamylase, and isomaltase in mixed mode with Ki values of 179.93, 176.38, 126.03, and 201.56 µg/mL, respectively. In silico screening of phytocompounds identified in NAFME indicated that hinokiflavone (HKF), pelargonidin-3-O-glucoside (PG), isorhamnetin-3-glucoside-7-rhamnoside (IGR), and petunidin-3-rutinoside (PR) showed better interactions with different subunits of human alpha-glucosidase, namely, N-terminal (Nt-MGAM and Nt-SI) and C-terminal (Ct-MGAM and Ct-SI). Molecular dynamics (MD) simulation, binding free energy study (molecular mechanics-generalized Born surface area [MM/GBSA]), and post-MD simulation studies (principal component analysis [PCA] and dynamic cross-correlation matrix [DCCM]) provided an in-depth understanding of these ligands' interactions with proteins. The overall efficacy of NAFME against oxidative stress and alpha-glucosidase in vitro is understood. Moreover, in silico analysis has shown the possible potential of HKF, PG, IGR, and PR to act as alpha-glucosidase inhibitors. Further studies on the antidiabetic potential of NAFME, HKF, PG, IGR, and PR in in vivo conditions are required to fully unveil the applicability of NAFME in the management of T2DM as a complementary medicine.</p>","PeriodicalId":9274,"journal":{"name":"Biotechnology and applied biochemistry","volume":" ","pages":""},"PeriodicalIF":3.2,"publicationDate":"2024-12-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142881287","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}