BMC Biotechnology最新文献

{"title":"Dual function of heterotrophic ammonia-oxidizing bacteria in facilitating maize compensatory growth under limited rewatering after drought.","authors":"Qiang Lv, Ruo-Yu Hao, Xiao-Ling Wang, Li-Ju Zhou, Lin Qi, Peng Song","doi":"10.1186/s12896-025-01006-z","DOIUrl":"10.1186/s12896-025-01006-z","url":null,"abstract":"<p><p>Water scarcity threatens global food security, making drought resilience in crops like maize crucial. In response to this challenge, this study investigates the potential of heterotrophic ammonia-oxidizing bacteria (HAOB) to enhance maize compensatory growth under post-drought limited rewatering conditions. Specifically, we focus on the dual mechanism of HAOB in modulating cytokinin synthesis and transport, aiming to develop an innovative agricultural biotechnology to support sustainable crop production. The S2_8_1 HAOB strain was used across two experiments. Experiment 1 investigated varying NO₃^- levels' effects on cytokinin translocation from roots to leaves under limited rewatering. Experiment 2 combined NO₃^- supplementation with HAOB inoculation to assess HAOB's twofold function in promoting compensatory growth under limited rewatering. The results showed that optimal NO₃^- levels (20-30 mmol·L^- 1 for limited rewatering) enhanced maize growth, root-to-shoot cytokinin translocation, and leaf cytokinin levels under limited rewatering. Notably, inoculation with HAOB outperformed these effects, demonstrating a more robust impact on cytokinin delivery and plant growth. This confirmed HAOB's twofold mechanism: Nitrification pathway - HAOB enhances rhizospheric NO₃⁻ availability, thereby stimulating cytokinin biosynthesis in roots and its translocation to leaves. Non-nitrification pathway - HAOB further promotes cytokinin translocation through mechanisms independent of soil NO₃⁻ increase. Sufficient rewatering increased rhizosphere nitrification rates, boosting root cytokinin translocation to leaves, thereby supported compensatory growth. Limited rewatering reduced rhizosphere nitrification, cytokinin translocation, and compensatory growth. However, HAOB overcame these constraints through its twofold function, enhancing cytokinin translocation and improving water use efficiency by more than fourfold, successfully promoting compensatory growth even under limited rewatering. Additionally, NO₃^- supplementation alleviated some limitations by increasing rhizosphere NO₃^-, but HAOB inoculation proved more effective, highlighting its superior role. This twofold function of HAOB strain significantly elevated cytokinin levels in leaves, supporting compensatory growth under limited rewatering. This biotechnology offers high agricultural potential, particularly in water-scarce regions, by improving drought resilience and yield stability.</p>","PeriodicalId":8905,"journal":{"name":"BMC Biotechnology","volume":"25 1","pages":"71"},"PeriodicalIF":3.5,"publicationDate":"2025-07-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12247422/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144616114","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Antimicrobial, antidiabetic, antiviral, and antioxidant activities of fucoidan extracted from the brown seaweed Padina pavonica.","authors":"Faiza M A Akl, Mostafa M El-Sheekh, Mofida E M Makhlof, Suzan I Ahmed","doi":"10.1186/s12896-025-01004-1","DOIUrl":"10.1186/s12896-025-01004-1","url":null,"abstract":"<p><p>Macroalgae are considered a promising biological source of active substances, and sulfated polysaccharides are among the most important compounds with potential applications. Here, fucoidan from Padina pavonica collected from the Rocky Bay of Abu Qir in Alexandria was analyzed for its ash, water, protein, sulfated groups, elemental content, total sugars, and uronic acid levels. Additionally, its monosaccharides were qualitatively and quantitatively determined using high-performance liquid chromatography for fucose detection, which is the main fingerprint of fucoidans. The isolated fucoidan was characterized using FTIR, scanning electron microscopy, EDX, and thermogravimetric analysis (TGA). Its biological activities, including antiviral, antioxidant, antimicrobial, and antidiabetic effects, were then evaluated. The yields of fucoidan, fucose, and sulfate were found to be 17.8 ± 0.23%, 34.45%, and 9.52 ± 0.19%, respectively. It inhibited HSV-1 with an inhibition percentage of 30.89 ± 0.84. The maximum Ferric Reducing Antioxidant Power (FRAP) value reached 81.82 ± 1.44% at 1000 µg/ml. Padina fucoidan (PF) showed the largest inhibition zone of 18 mm against Methicillin-Resistant Staphylococcus aureus (MARSA) (ATCC 4330) with an MIC of 1.25 mg/L. It also demonstrated a promising inhibitory effect on α-Amylase enzyme, reaching 75.69 ± 1.05 at a concentration of 1000 µg/ml. We conclude that Padina pavonica is an excellent producer of fucoidan, with a significant sulfate content that enhances its biological activities, especially its antidiabetic properties.</p>","PeriodicalId":8905,"journal":{"name":"BMC Biotechnology","volume":"25 1","pages":"70"},"PeriodicalIF":3.5,"publicationDate":"2025-07-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12243346/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144599158","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Carotenoid metabolomic and transcriptomic analyses provide insights into the flower color transition in Lonicera macranthoides.","authors":"Ling Ling Lv, Li Yun Li, Jian Hui Pi","doi":"10.1186/s12896-025-01007-y","DOIUrl":"10.1186/s12896-025-01007-y","url":null,"abstract":"<p><strong>Background: </strong>In Lonicera macranthoides (L. macranthoides), the role of carotenoids in the flower color transition remains unclear. In this study, at the four flower developmental stages of green flower bud (GB), white flower bud (WB), white flower (WF) and golden flower (GF) in L.macranthoides, combined transcriptomic and carotenoid metabolomic analyses was performed to clarify the changes of carotenoid content and the expressions of genes related to carotenoid biosynthesis.</p><p><strong>Results: </strong>A total of sixteen carotenoids, including 5 carotenes and 11 xanthophylls, were detected and quantified from the all samples. The content of 16 carotenoids was found to decrease first and reach the lowest level at WF, then dramatically increase at GF. At GB, the carotenoid content was the highest and the top three carotenoids in content were lutein, zeaxanthin and violaxanthin. At WB and WF, the carotenoid contents were relatively low, and the buds or flowers appeared white. At GF, β-carotene and violaxanthin were obviously dominant, accounting for 64.95% of the content of 14 detectable carotenoids, and they might be the major contributors to yellow color at GF. The expressions of differentially expressed genes indicated that with the development of flowers, the biosynthesis of carotenoids shifted from α-carotene branch to β-carotene branch.</p><p><strong>Conclusion: </strong>The findings are beneficial to genetic improvement of varieties in L.macranthoides by increasing the carotenoids content in flowers.</p>","PeriodicalId":8905,"journal":{"name":"BMC Biotechnology","volume":"25 1","pages":"69"},"PeriodicalIF":3.5,"publicationDate":"2025-07-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12243273/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144599159","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Bioprocessing and characterization of thermostable phytase from Aspergillus terreus, an endophyte of Catharanthus roseus, with a potential activity to hydrolyze phytic acid in wheat bran.","authors":"Marwa A Yassin, Eman H Mohsen, Nelly M George, Mohamed A Marawan, Ashraf S A El-Sayed, Marwa M El-Demerdash","doi":"10.1186/s12896-025-00988-0","DOIUrl":"10.1186/s12896-025-00988-0","url":null,"abstract":"<p><p>Phytic acid is one of the common anti-nutritional factors in animal feeds, due to its chelating activity of metal ions and amino acids, so, phytase has been used for increasing the nutritional value of the animal feeds by releasing phosphorous. The stability and catalytic efficiency of this enzyme are the major challenges, so, the objective of this study was to purify and characterize phytase with relatively unique biochemical properties. Among the recovered fungal endophytes of Catharanthus roseus, Aspergillus terreus EFBL-AS PV412881.1 was recognized as the most potent phytase producing isolate. Upon nutritional optimization with the face-centered central composite design (FCCD), the productivity of phytase by A. terreus grown on wheat bran amended with 0.2% NaNO₃ and 0.4 % yeast extract, under solid state fermentation, was increased into 36.3 μmol/mg/min. Phytase of A. terreus was purified to its molecular homogeneity by gel-filtration and ion-exchange chromatography, with 3.48 purification folds (125 μmol/mg/min). The purified enzyme had a molecular subunit 85 kDa by denaturing-PAGE, with highest activity at reaction temperature 37-40 °C, and reaction pH 7.0. The T_1/2 of A. terreus phytase was 124.5, 5.2 and 3.8 h, at 4, 40, and 50 °C, respectively. The thermal denaturation rate (Kr) was 0.095 ×10^-3, 0.27 × 10^-3 and 0.292 ×10^-3/min at 20, 40, and 50 °C, respectively. The enzyme was slightly inhibited by Ca²⁺ ions, unlike the resistance to various cations. The concentration of phytic acid of wheat bran was reduced by about 6.5 folds upon phytase treatment, ensuring the feasibility of this enzyme in the animal feed application. From the molecular docking analysis, phytase from A. terreus had a higher affinity to hydrolyze phytic acid, with binding energy - 7.1 kcal/mol, compared to that of A. niger and P. pinophilum (-6.7 kcal/mol), ensuring the stability of the interaction.</p>","PeriodicalId":8905,"journal":{"name":"BMC Biotechnology","volume":"25 1","pages":"68"},"PeriodicalIF":3.5,"publicationDate":"2025-07-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12239475/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144590359","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"CDCP1 promotes the malignant phenotypes of nasopharyngeal carcinoma via the Wnt/β-catenin signaling pathway.","authors":"Guoliang Bie, Shuang Cheng, Weiping Huang, Zhongpu Yin, Jianzhi Liu","doi":"10.1186/s12896-025-01001-4","DOIUrl":"10.1186/s12896-025-01001-4","url":null,"abstract":"<p><strong>Background: </strong>CUB domain-containing protein 1 (CDCP1), a type I transmembrane glycoprotein, is abundantly expressed in various cancers. However, its role and mechanism in nasopharyngeal carcinoma (NPC) remain ambiguous.</p><p><strong>Methods: </strong>The UALCAN and GEPIA databases were analyzed to explore CDCP1 expression and survival prognosis in head and neck squamous cell carcinoma (HNSC) patients. Fifteen pairs of NPC tissues and adjacent normal tissues were collected for CDCP1 expression analysis. CCK-8 assays, flow cytometry, and transwell assays were performed on NPC cell lines (C666-1, 5-8 F, and HONE-1). The impact of GSK-3β inhibitor LiCl on C666-1 cells after CDCP1 knockdown was investigated. A C666-1 xenograft model was established for in vivo validation.</p><p><strong>Results: </strong>CDCP1 was overexpressed in HNSC patients, and elevated CDCP1 correlated with poor survival. NPC tissues confirmed CDCP1 upregulation compared to normal tissues. CDCP1 knockdown in C666-1 and 5-8 F cells inhibited proliferation, migration, invasion, and promoted apoptosis, while LiCl partially reversed these effects. In vivo, CDCP1 silencing suppressed tumor growth, downregulated PCNA, Wnt3a, β-catenin, and p-GSK-3β, and upregulated cleaved caspase-3 and E-cadherin. CDCP1 overexpression in HONE-1 cells produced opposing effects.</p><p><strong>Conclusions: </strong>In summary, CDCP1 promotes NPC progression via the Wnt/β-catenin pathway, suggesting its potential as a therapeutic target.</p><p><strong>Clinical trial number: </strong>Not applicable.</p>","PeriodicalId":8905,"journal":{"name":"BMC Biotechnology","volume":"25 1","pages":"67"},"PeriodicalIF":3.5,"publicationDate":"2025-07-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12235957/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144582963","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Unlocking the antioxidant and antibacterial potential of exopolysaccharides produced by endophytic fungi (Aspergillus fumigatus and Preussia isabellae) isolated from Tanzania's mangroves.","authors":"Bhoke Marwa Nyaisaba, Rose Justus Masalu, Hawa Myovela, Cyprian Beda Mpinda","doi":"10.1186/s12896-025-01005-0","DOIUrl":"10.1186/s12896-025-01005-0","url":null,"abstract":"","PeriodicalId":8905,"journal":{"name":"BMC Biotechnology","volume":"25 1","pages":"66"},"PeriodicalIF":3.5,"publicationDate":"2025-07-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12229027/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144567025","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Scientific and technological challenges of recombinant egg protein production.","authors":"Franziska Beck, Philipp Noll, Ute Schweiggert-Weisz, Marius Henkel","doi":"10.1186/s12896-025-01002-3","DOIUrl":"10.1186/s12896-025-01002-3","url":null,"abstract":"<p><p>Eggs are among the most widely consumed and versatile animal-derived foods and are valued for their exceptional nutritional and functional properties. However, conventional egg production is associated with significant environmental, ethical, and health concerns, increasing the global consumer demand for sustainable animal protein. To address these challenges, recombinant egg protein production through precision fermentation has emerged as a promising alternative. Yet, this alternative production path is still in its infancy, and current efforts in research have not yet led to a widespread adoption of recombinant egg protein. This review provides an overview of the bioprocesses used to produce recombinant egg proteins, highlighting their nutritional, bio-functional, and techno-functional significance. The current state of the art of recombinant egg protein production is presented, with a comparison of different microbial expression hosts in terms of suitability and associated challenges. Only six egg proteins were reported to be expressed at laboratory scale, including ovalbumin (3.7 g/L with Escherichia coli EcN) and ovomucoid (3.2 g/L with Komagataella phaffii). The realization of large-scale production of functional egg proteins remains challenging. These challenges include posttranslational modifications, achieving functionality and cost parity to natural egg proteins, efficient resource bioconversion, and optimizing the bioprocess chain (upstream, bioproduction, and downstream processes). This requires further improvements and research to increase protein titers, space-time yields, and production rates. Nevertheless, recombinant egg protein produced via precision fermentation holds great promise as a functional food ingredient. With further advancements, this approach could contribute to global protein demand, enhance food security, and strengthen food system resilience while providing a more sustainable and ethical alternative to conventional egg production.</p>","PeriodicalId":8905,"journal":{"name":"BMC Biotechnology","volume":"25 1","pages":"65"},"PeriodicalIF":3.5,"publicationDate":"2025-07-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12224593/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144551847","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Evaluating the anticancer efficacy of Chara vulgaris ethanolic extract and selenium nanoformulation in Ehrlich carcinoma mice: role of autophagy and apoptosis.","authors":"Maha Alsunbul, Thanaa A El-Masry, Maisra M El-Bouseary, Enas I El Zahaby, Mostafa M El-Sheekh, Mohamed M S Gaballa, Eman Wahsh, Heba Kamel Badawy, Jawaher Abdullah Alamoudi, Reem ALQahtani, Naifa Alenazi, Maysa M F El-Nagar","doi":"10.1186/s12896-025-00998-y","DOIUrl":"10.1186/s12896-025-00998-y","url":null,"abstract":"<p><strong>Background: </strong>Bioactive compounds with a wide range of chemical compositions and biological functions are found in the Chlorophyceae family. The present work investigated the anticancer effect of ethanolic extract from Chara vulgaris (C. vulgaris) and its selenium nanoformulation (CvSeNPs) against solid Ehrlich carcinoma (SEC).</p><p><strong>Methods: </strong>Gas chromatography-mass spectroscopy was used to analyze C. vulgaris, and many analytical methods were used to characterize the biosynthesized CvSeNPs, including zeta potential, particle size, polydispersity index (PDI), SEM, TEM, and FTIR. In addition, mice with SEC were randomly assigned to seven equal groups (n = 6) to investigate possible mechanisms behind the antitumor activity. Group 1: Tumor control, group 2: Tamoxifen (10 mg/kg), group 3: Free SeNPs, group 4: C. vulgaris (25 mg/kg), group 5: C. vulgaris (50 mg/kg), group 6: 25 mg/kg CvSeNPs, group 7: 50 mg/kg CvSeNPs.</p><p><strong>Results: </strong>Gas chromatography-mass spectroscopy analysis of the ethanolic extract of C. vulgaris revealed the presence of several bioactive chemicals that may promote anticancer activity. Protein levels of TNF-α, NF-кB, VEGF, and Bcl-2 were suppressed in the CvSeNPs group (50 mg/kg), whereas those of caspase-3, BAX, P53, P62, LC3, and Beclin1 were increased. Additionally, CvSeNPs significantly exceeded similar dosages of free extract in terms of caspase-3, BAX, Bcl-2, P53, TNF-α, NF-кB, VEGF, P62, LC3, and Beclin1 gene expression.</p><p><strong>Conclusion: </strong>The CvSeNPs mediated their positive anticancer impact, which manifested as a decrease in tumor volume and an improvement in overall survival rate, by promoting autophagy, apoptosis, and lowering inflammation.</p>","PeriodicalId":8905,"journal":{"name":"BMC Biotechnology","volume":"25 1","pages":"53"},"PeriodicalIF":3.5,"publicationDate":"2025-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12211136/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144537987","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"An engineered self-cleavage fusion system for the production of chimaera spider silk proteins.","authors":"Yongqin Su, Ke Zheng, Cheng Cheng","doi":"10.1186/s12896-025-00987-1","DOIUrl":"10.1186/s12896-025-00987-1","url":null,"abstract":"<p><strong>Background: </strong>Spidroins are well-known for their exceptional mechanical properties, which have inspired extensive research and applications across various fields. Large-scale production of spidroin continues to face major challenges due to the complexity involved in inducing host organisms to express full-length spidroins. This process requires advanced techniques and has issues such as plasmid instability and potential misfolding.</p><p><strong>Results: </strong>In this study, we developed a novel expression system by combining a fusion tag with a self-cleavage intein, enabling the convenient expression of three chimeric spidroins with varying numbers of repetitive units. After optimising expression conditions, NT2RepCT, NT4RepCT and NT6RepCT spidroins were obtained in soluble form, with yields of 266, 135 and 125 mg/L, respectively. All three spidroins exhibited an increased β-sheet content with increased numbers of repetitive units during transition from soluble to dry state. In terms of nanofibril morphologies, the self-assemblies of NT4RepCT and NT6RepCT closely resemble those of native silk proteins.</p><p><strong>Conclusion: </strong>This study can serve as a reference for preparation of high-performance spider silk materials and soluble expression of proteins, such as collagen, and as a foundation for preparation of other structurally complex polymer materials.</p>","PeriodicalId":8905,"journal":{"name":"BMC Biotechnology","volume":"25 1","pages":"56"},"PeriodicalIF":3.5,"publicationDate":"2025-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12218096/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144537902","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Comparison of active site mutations at subsite + 2 of Anoxybacillus ayderensis A9 β-glucosidase for hydrolysis of pNPG and polydatin.","authors":"Numan Saleh Zada, Ali Osman Belduz, Abdulrahman H Alessa, Halil Ibrahim Güler, Mine Karaoğlan, Malik Badshah, Aamer Ali Shah, Aasia Kalsoom, Samiullah Khan","doi":"10.1186/s12896-025-00984-4","DOIUrl":"10.1186/s12896-025-00984-4","url":null,"abstract":"<p><p>β-glucosidase from Anoxybacillus ayderensis A9 (BglA9) is a potent enzyme for enzymatic hydrolysis of polydatin to resveratrol. Based on structural and bioinformatics analysis an area near + 2 subsite of the active site pocket of BglA9 was selected and single point mutations were introduced with the aim to enhance the catalytic efficiency of the enzyme towards pNPG and polydatin. The active site region selected for mutations is non-conserved between different glycoside hydrolase family 1 (GH1) enzymes and is located at the end of β-strand 6. The changes introduced in the active site residues were L221S, N222S and G226Q. The E. coli BL21 (DE3) cells were used for the expression of mutant proteins and purification was achieved by Ni-NTA column chromatography. The thermal and pH stability was retained in all the mutants. The proteins with mutated residue resulted in variations in K_m and k_cat/K_m (catalytic efficiency) values. The K_m values of mutants for pNPG and polydatin were lowered, indicating a better enzyme-substrate complex, while variations in k_cat/K_m values were observed for both substrates. The docking analysis supported these observations and by comparing binding energies; the mutant N222S showed the best docked complex. This investigation suggests that the + 2 subsite of BglA9 is an interesting area to be mutated and changes in amino acid residues at this site can influence both K_m and catalytic efficiency. The deglycosylated derivates were also compared for their antioxidant activities and showed enhanced antioxidant potential as compared to glycoside measured by DPPH assay.</p>","PeriodicalId":8905,"journal":{"name":"BMC Biotechnology","volume":"25 1","pages":"52"},"PeriodicalIF":3.5,"publicationDate":"2025-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12211328/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144537986","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}