Biotechnology Letters最新文献

{"title":"Clinical and mechanistic insights into biomedical application of Se-enriched probiotics and biogenic selenium nanoparticles.","authors":"Farshid Ataollahi, Bagher Amirheidari, Zohreh Amirheidari, Mahshid Ataollahi","doi":"10.1007/s10529-024-03559-z","DOIUrl":"https://doi.org/10.1007/s10529-024-03559-z","url":null,"abstract":"<p><p>Selenium is an essential element with various industrial and medical applications, hence the current considerable attention towards the genesis and utilization of SeNPs. SeNPs and other nanoparticles could be achieved via physical and chemical methods, but these methods would not only require expensive equipment and specific reagents but are also not always environment friendly. Biogenesis of SeNPs could therefore be considered as a less troublesome alternative, which opens an excellent window to the selenium and nanoparticles' world. bSeNPs have proved to exert higher bioavailability, lower toxicity, and broader utility as compared to their non-bio counterparts. Many researchers have reported promising features of bSeNP such as anti-oxidant and anti-inflammatory, in vitro and in vivo. Considering this, bSeNPs have been tried as effective agents for health disorders, especially as constituents of probiotics. This article briefly reviews selenium, selenium nanoparticles, Se-enriched probiotics, and bSeNPs' usage in an array of health disorders. Obviously, there are very many articles on bSeNPs, but we wanted to summarize studies on prominent bSeNPs features published in the twenty-first century. This review is hoped to give an outlook to researchers for their future investigations, ultimately serving better care of health disorders.</p>","PeriodicalId":8929,"journal":{"name":"Biotechnology Letters","volume":"47 1","pages":"18"},"PeriodicalIF":2.0,"publicationDate":"2025-01-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142999207","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":"Comparative assessment of chondral defect repair using human bone marrow- and adipose tissue-derived mesenchymal stem cells, adult and foetal articular cartilage-derived chondrocytes, and chondroprogenitors: an ex-vivo model.","authors":"Elizabeth Vinod, Ganesh Parasuraman, Jeya Lisha J, Jithu James Varghese, Abel Livingston, Grace Rebekah, Deepak Vinod Francis, Solomon Sathishkumar, Alfred Job Daniel, Boopalan Ramasamy","doi":"10.1007/s10529-024-03558-0","DOIUrl":"https://doi.org/10.1007/s10529-024-03558-0","url":null,"abstract":"<p><strong>Purpose: </strong>Cartilage repair necessitates adjunct therapies such as cell-based approaches, which commonly use MSCs and chondrocytes but is limited by the formation of fibro-hyaline cartilage. Articular cartilage-derived chondroprogenitors(CPs) offer promise in overcoming this, as they exhibit higher chondrogenic and lower hypertrophic phenotypes. The study aimed to compare the efficacy of various cell types derived from adult and foetal cartilage suspended in platelet-rich plasma(PRP) in repairing chondral defects in an Ex-vivo Osteochondral Unit(OCU) model.</p><p><strong>Methods: </strong>In-vitro characterization of the cells included growth kinetics, FACS, qRT-PCR, and multilineage differentiation potential using histology and GAG analysis. Ex-vivo human OCUs with chondral defects containing the different cells in PRP were cultured and subjected to analysis for matrix and collagen staining.</p><p><strong>Results: </strong>The ex-vivo OCU analysis, in terms of defect repair, showed that adult chondrocytes, sorted-CPs, and foetal MCPs displayed better host integration and filling. The In-vitro analysis of adult chondrocytes displayed greater chondrogenic genes ACAN and COL2A1 expression, with sorted-CPs also showing higher levels of ACAN. In terms of accumulation of extracellular matrix uptake evident by Safranin O staining and collagen type II fibrillar uptake, the AD-MSCs, BM-MSCs, and sorted CPs outperformed the other groups. BM-MSCs also showed corroborative higher CD146 levels, however, the gene analysis of the AD-MSCs showed a high hypertrophic tendency in terms of its COL1A1 and RUNX2 expression.</p><p><strong>Conclusion: </strong>Sorted chondroprogenitors outperformed both in terms of filling and hyaline-like repair, with AD-MSC and BM-MSC groups also achieving functional cartilage of a hyaline nature, warranting further evaluation using in-vivo and clinical studies.</p>","PeriodicalId":8929,"journal":{"name":"Biotechnology Letters","volume":"47 1","pages":"17"},"PeriodicalIF":2.0,"publicationDate":"2025-01-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142943759","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":"Application of aqueous two-phase extraction for separation and purification of various adeno-associated viruses.","authors":"Xiao-Qian Fu, Hui-Yi Leong, Liang-Zhi Qiao, Jia-Nan Zhou, Wei Hu, Shan-Jing Yao, Dong-Qiang Lin","doi":"10.1007/s10529-024-03555-3","DOIUrl":"https://doi.org/10.1007/s10529-024-03555-3","url":null,"abstract":"<p><strong>Objective: </strong>Adeno-associated viruses (AAVs) are widely used as gene therapy vectors due to their safety, stability, and long-term expression characteristics. The objective of this work is to develop an aqueous two-phase system (ATPS) as a universal platform for the separation and purification of AAVs.</p><p><strong>Results: </strong>This study utilized polyethylene glycol (PEG)/salt ATPSs to separate and purify various AAV serotypes, including AAV5, AAV8, and AAV9, which focusing on serotype-specific performance and partial empty capsid removal. The results showed that all the AAV serotypes were mainly enriched in the interphase of ATPS, with achieving high recovery (> 95%) and impurity removal (> 95%). The PEG/sodium citrate ATPS was serotype-independent, but the process optimization of component concentrations for each serotype was necessary to attain the best performance. Notably, a single-step aqueous two-phase extraction also demonstrated the ability to remove some amount of empty capsids from the crude cell lysate, with removal rate ranging from 4 to 25%.</p><p><strong>Conclusions: </strong>The results demonstrated the practical applicability of PEG/sodium citrate ATPS in separating and purifying different AAV serotypes, which addressing key challenges in gene therapy vector production.</p>","PeriodicalId":8929,"journal":{"name":"Biotechnology Letters","volume":"47 1","pages":"16"},"PeriodicalIF":2.0,"publicationDate":"2025-01-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142943757","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":"Coselection of BAC for Escherichia coli chromosomal DNA multiplex automated genome engineering.","authors":"Junyu Wang, Hong Wang, Jiamei Wang, Guangdong Shang","doi":"10.1007/s10529-024-03554-4","DOIUrl":"https://doi.org/10.1007/s10529-024-03554-4","url":null,"abstract":"<p><p>Recombineering (recombination-mediated genetic engineering) is a powerful strategy for bacterial genomic DNA and plasmid DNA modifications. CoS-MAGE improved over MAGE (multiplex automated genome engineering) by co-electroporation of an antibiotic resistance repair oligo along with the oligos for modification of the Escherichia coli chromosome. After several cycles of recombineering, the sub-population of mutants were selected among the antibiotic resistant colonies. However, a pre-generated strain with mutS deletion and multiple inactivated antibiotic resistance genes integration is required. Herein, CoS-MAGE was modified by employing a single copy BAC vector harboring a bla-mkan cassette and a Red helper vector cloned with dominant mutL E32K, thus bypassing the utilization of the pre-generated strain. The proof-of-concept of the new strategy, CoS-BAC-MAGE, was demonstrated via the mutation of non-essential genes, essential genes, and AT rich regions of the wild type strain E. coli MG1655. With this system, an editing efficiency of 60% was realized. Furthermore, by toggling between two antibiotic resistance genes (one active, the other defective) on the BAC, sequential mutations were achieved without the requirement of BAC vector elimination and re-transformation. Via CoS-BAC-MAGE, simultaneously mutations of three sites were obtained in a day. We envision that CoS-BAC-MAGE will be a practical improvement for the generation of chromosomal mutations using the Cos-MAGE approach.</p>","PeriodicalId":8929,"journal":{"name":"Biotechnology Letters","volume":"47 1","pages":"14"},"PeriodicalIF":2.0,"publicationDate":"2024-12-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142891788","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":"An HRP-integrated CRISPR/Cas12a biosensor towards chair-side diagnosis for Porphyromonas gingivalis.","authors":"Ningning Pi, Rong Xiang, Lu Zhu, Yi Li, Xuan Wu","doi":"10.1007/s10529-024-03556-2","DOIUrl":"https://doi.org/10.1007/s10529-024-03556-2","url":null,"abstract":"<p><p>Rapid diagnostic tools for Porphyromonas gingivalis (Pg), the primary microorganism responsible for the development of periodontitis, particularly those designed for chair-side applications, could provide substantial health benefits to patients. To address this issue, we developed a CRISPR/Cas12a-based rapid Pg detection method. Dual-gRNA and hairpin reporter strategies were employed to enhance CRISPR/Cas12a reaction efficiency. By modifying the hairpin reporter with HRP, the pre-amplification-free HRP-CRISPR/Cas12a reaction was enabled to produce a colorimetric output, amplifying the detection signal. This method achieved high sensitivity (as low as 33 CFU) without the risk of aerosol contamination from pre-amplification. When testing clinical samples, the method showed high consistency with the reference RT-PCR. Furthermore, compared with RT-PCR, this method only requires room temperature operation, is simpler, and has a shorter detection time of about 35 min. In conclusion, the pre-amplification-free HRP-integrated CRISPR/Cas12a detection method requires no complex equipment, making it an ideal, end-user-friendly approach for chair-side Pg detection.</p>","PeriodicalId":8929,"journal":{"name":"Biotechnology Letters","volume":"47 1","pages":"15"},"PeriodicalIF":2.0,"publicationDate":"2024-12-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142891784","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":"Comparative proteomic analysis to annotate the structural association of the hypothetical proteins from the conserved domain of P. aeruginosa as novel vaccine candidates.","authors":"Prajval Tenginakai, Samiksha Bhor, Fathimathuz Zehra Waasia, Sameer Sharma, Susha Dinesh","doi":"10.1007/s10529-024-03546-4","DOIUrl":"https://doi.org/10.1007/s10529-024-03546-4","url":null,"abstract":"<p><strong>Objectives: </strong>Pseudomonas aeruginosa, identified as an ESKAPE pathogen, contributes to severe clinical diseases worldwide and despite its prevalence an effective vaccine or treatment remains elusive. Numerous computational methods are being employed to target hypothetical proteins (HPs). Presently, no studies have predicted multi-epitope vaccines for these HPs.</p><p><strong>Results: </strong>Totally, 877 HPs from P. aeruginosa were included in the study and the data showcased here illustrate a methodical approach to prioritize the proteome by employing diverse comparative proteomics. The study employed physicochemical property assessment and conserved domain analysis to identify stable and immunologically pertinent proteins for epitope prediction. The VaxiJen2.0 antigenicity assessment aided in epitope selection, contributing to the foundational steps in vaccine development by predicting T-cell and B-cell epitopes. Potential T and B cell epitopes with high antigenicity, non-toxic categorization, and robust binding affinities were identified in the investigation. The periplasmic HP WP_132813935.1 was predicted as conserved, stable, and soluble. The T-cell peptide RTSMRALAY and the B-cell peptide MPVYLYLM were predicted to be probable non-allergen and demonstrated strong binding with MHC class I allele HLA-C*03:03.</p><p><strong>Conclusions: </strong>This research provides a comprehensive approach to predict T and B cell epitopes for conditions associated with P. aeruginosa, offering a candidate pool for tailored vaccine development. However, the efficacy of these epitopes in vaccine development necessitates clinical validation and testing for confirmation.</p>","PeriodicalId":8929,"journal":{"name":"Biotechnology Letters","volume":"47 1","pages":"13"},"PeriodicalIF":2.0,"publicationDate":"2024-12-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142862957","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":"Biocontrol toxicity of Trichoderma harzianum (Hypocreales: Hypocreaceae) derived chemical molecules against malarial mosquito Anopheles stephensi with molecular docking studies.","authors":"Perumal Vivekanandhan, Abdullah A Alarfaj, Saleh Alfarraj, Mohammad Javed Ansari, Chinnaperumal Kamaraj","doi":"10.1007/s10529-024-03542-8","DOIUrl":"https://doi.org/10.1007/s10529-024-03542-8","url":null,"abstract":"<p><p>In this study, the crude chemical constituents extracted from Trichoderma harzianum and their toxicity were evaluated against the larvae, pupae, and adults of Anopheles stephensi at 24 and 48 h post-treatment. Additionally, the chemical constituents of the crude extracts were identified using gas chromatography-mass spectrometry (GC-MS) analysis, and their ability to bind with target proteins was confirmed through molecular docking studies. The results clearly demonstrated that the chemical compounds from T. harzianum exhibited promising mortality rates in larvae (98.66%), pupae (92%), and adult mosquitoes (81.33%) of A. stephensi 48 h after treatment. The study assessed the impact of crude extracts on insect enzymes 24 h post treatment, revealing significant alterations: a reduction in catalase activity and an increase in glutathione S-transferase levels compared to the control group. The treatment with crude chemical extracts resulted in mortality rates of 37.33% and 52% at 24 and 48 h, respectively, on Artemia salina , indicating minimal effects. After 48 h, the crude extract exhibited minimal toxicity on Eudrilus eugeniae, with a recorded mortality rate of 15% after 48 h. GC-MS analysis of T. harzianum-derived crude extracts identified ten major chemical constituents. Among these, chemicals, 2,4-bis(1,1-dimethylethyl) phenol (19.02%) was recognized as the predominant chemical component. This 2,4-bis(1,1-dimethylethyl) phenol molecule demonstrates a high binding affinity with target proteins, which is a key factor contributing to its insecticidal activity. This study concludes that the chemical constituents derived from T. harzianum are promising candidates for an eco-friendly, effective, and target-specific alternative control method for A. stephensi mosquitos.</p>","PeriodicalId":8929,"journal":{"name":"Biotechnology Letters","volume":"47 1","pages":"12"},"PeriodicalIF":2.0,"publicationDate":"2024-12-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142817042","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":"A recombinant L-threonine aldolase with high catalytic efficiency for the asymmetric synthesis of L-threo-phenylserine and L-threo-4-fluorophenylserine.","authors":"Bijing Lei, Wan Jiang, Jinsong Ma, Caiyun Wang, Yinping Pan, Zhi Zhang, Bochu Wang, Jian Guo, Na Qi","doi":"10.1007/s10529-024-03553-5","DOIUrl":"https://doi.org/10.1007/s10529-024-03553-5","url":null,"abstract":"<p><strong>Objectives: </strong>To develop robust variants of L-threonine aldolases (L-TAs), potent catalysts for synthesizing asymmetric β-hydroxy-α-amino acids, it is necessary to identify critical residues beyond the known active site residues.</p><p><strong>Results: </strong>Through virtual screening, a neglected residue Asn305, was identified as critical for catalytic efficiency. Subsequent site-saturation mutagenesis led to a potent variant N305R which exhibited excellent conversions of 88%_conv (87%_de) and 80%_conv (94%_de) for the synthesis of L-threo-phenylserine and L-threo-4-fluorophenylserine respectively. This variant not only outperformed the template enzyme, but also represented a promising L-TA for synthesizing the two β-hydroxy-α-amino acids. It was suggested that Arg305 of the variant N305R generated strong cation-arene interaction and electrostatic force with the intermediates, leading to strengthened binding, enhanced L-threo favored orientation and wider entrance.</p><p><strong>Conclusions: </strong>Our work not only provided an excellent variant N305R, but also suggested the crucial function of a neglected residue Asn305, which offered valuable experiences for other L-TA studies.</p>","PeriodicalId":8929,"journal":{"name":"Biotechnology Letters","volume":"47 1","pages":"11"},"PeriodicalIF":2.0,"publicationDate":"2024-12-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142799356","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":"Identification, heterologous expression, and characterisation of β-1,3-xylanase BcXyn26B from human gut bacterium Bacteroides cellulosilyticus WH2.","authors":"Sanae Hori, Fumiyoshi Okazaki","doi":"10.1007/s10529-024-03547-3","DOIUrl":"10.1007/s10529-024-03547-3","url":null,"abstract":"<p><p>The cell walls of red and green algae contain β-1,3-xylan, which is hydrolysed by the endo-type enzyme β-1,3-xylanase. Notably, only marine-bacteria-derived β-1,3-xylanases have been functionally characterised to date. In this study, we characterised the enzymatic properties of a potential β-1,3-xylanase (BcXyn26B) derived from the human gut bacterium, Bacteroides cellulosilyticus WH2. The codon optimized BcXyn26B gene was synthesised and expressed in Escherichia coli BL21(DE3). The recombinant protein was purified by a two-step purification process using Ni-affinity chromatography followed by anion exchange chromatography, and its enzymatic properties were characterised. The recombinant BcXyn26B exhibited specific hydrolytic activity against β-1,3-xylan and released various β-1,3-xylooligosaccharides, with β-1,3-xylobiose as the primary product. The optimum reaction temperature was 50 °C, higher than that for other enzymes derived from marine bacteria. This study represents the first report on the identification, heterologous expression, and characterisation of β-1,3-xylanase from human gut microbes. Notably, the substrate specificity of BcXyn26B indicates that human gut Bacteroides species possess an unknown β-1,3-xylan utilisation system.</p>","PeriodicalId":8929,"journal":{"name":"Biotechnology Letters","volume":"47 1","pages":"10"},"PeriodicalIF":2.0,"publicationDate":"2024-12-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11611983/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142765745","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Homologous expression of Aspergillus niger α-L-rhamnosidase and its application in enzymatic debittering of Ougan juice.","authors":"Fei Zhang, Xue Wang, Lixia Pan, Zhao Wang, Jianyong Zheng","doi":"10.1007/s10529-024-03531-x","DOIUrl":"10.1007/s10529-024-03531-x","url":null,"abstract":"<p><p>The α-L-rhamnosidase (rha1) gene was homologously expressed in Aspergillus niger strains CCTCC 206047 and CCTCC 206047ΔpyrG, using hygromycin B and auxotrophic as selection markers. The engineered A. niger strains RHA001-1 and RHA003-1 were screened, yielding α-L-rhamnosidase activities of 20.81 ± 0.56 U/mL and 15.35 ± 0.87 U/mL, respectively. The copy numbers of the rha1 gene in strains RHA001-1 and RHA003-1 were found to be 18 and 14, respectively. Correlation analysis between copy number and enzyme activity in the A. niger strains revealed that α-L-rhamnosidase activity increased with the copy number of the rha1 gene. Recombinant α-L-rhamnosidase was utilized for the enzymatic debittering of Ougan juice, and its process conditions were optimized. Furthermore, the primary bitter substance neohesperidin (2.22 g/L) in Ougan juice was converted into hesperetin 7-O-glucoside (1.47 g/L) and hesperidin (0.143 g/L). This study presents a novel approach for the production of food-grade α-L-rhamnosidase and establishes a technical foundation for its application in the beverage industry.</p>","PeriodicalId":8929,"journal":{"name":"Biotechnology Letters","volume":" ","pages":"1187-1198"},"PeriodicalIF":2.0,"publicationDate":"2024-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142131748","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}