Biotechnology Letters最新文献

{"title":"Efficient RNA interference method by feeding in Brachionus plicatilis (Rotifera).","authors":"Yu Zhang, Dongqi Kan, Yang Zhou, Hairong Lian, Lingling Ge, Jing Shen, Zhongqi Dai, Yan Shi, Cui Han, Xiaojie Liu, Jiaxin Yang","doi":"10.1007/s10529-024-03524-w","DOIUrl":"10.1007/s10529-024-03524-w","url":null,"abstract":"<p><p>Rotifers are small, ubiquitous invertebrate animals found throughout the world and have emerged as a promising model system for studying molecular mechanisms in the fields of experimental ecology, aquatic toxicology, and geroscience. However, the lack of efficient gene expression manipulation techniques has hindered the study of rotifers. In this study, we used the L4440 plasmid with two reverse-oriented T7 promoters, along with RNase-deficient E. coli HT115, to efficiently produce dsRNA and thereby present an efficient feeding-based RNAi method in Brachionus plicatilis. We targeted Bp-Ku70 & Ku80, key proteins in the DNA double-strand breaks repair pathway, and then subjected rotifers to UV radiation. We found that the mRNA expression, fecundity, as well as survival rate diminished significantly as a result of RNAi. Overall, our results demonstrate that the feeding-based RNAi method is a simple and efficient tool for gene knockdown in B. plicatilis, advancing their use as a model organism for biological research.</p>","PeriodicalId":8929,"journal":{"name":"Biotechnology Letters","volume":" ","pages":"961-971"},"PeriodicalIF":2.0,"publicationDate":"2024-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142131747","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":"Characterization, immobilization and evaluation of anti-Pseudomonas aeruginosa biofilm activity of alginate lyase from marine bacterium, Enterobacter tabaci RAU2C.","authors":"Ramya Petchimuthu, Krishnan Sundar, Vanavil Balakrishnan","doi":"10.1007/s10529-024-03551-7","DOIUrl":"10.1007/s10529-024-03551-7","url":null,"abstract":"<p><p>Alginate lyases have the potential to be used as a therapeutic agent for P. aeruginosa infections. The present work was focused on the characterization of free and immobilized alginate lyase produced by marine bacteria, Enterobacter tabaci RAU2C isolated previously in the laboratory for alginate lyase production and exploring the potential of alginate lyase as an anti-biofilm agent against the P. aeruginosa biofilm. RAU2C alginate lyase was immobilized using an epoxy-activated curdlan matrix by three different methods. Further, the free and immobilized were characterized for its optimal pH and temperature. The effect of alginate concentration on alginate lyase activity was assessed and the kinetic parameters were evaluated. The anti-biofilm activity of the crude alginate lyase was studied using biofilm inhibition and disruption assays in microtiter plates with crystal violet. The biofilm disruption by RAU2C alginate lyase was also ascertained by microscopic analysis. The immobilization matrix prepared using method 3 had a better binding capacity compared to other methods. Both soluble and immobilized alginate lyase exhibited optimal activity at 37 °C and pH 7.0. K_m and V_max of soluble and immobilized alginate lyase were found to be 3.38 mg/mL, 22.98 mg/mL min and 3.67 mg/mL and 26.59 mg/mL min respectively. Both microtiter assay and microscopic analysis confirmed the prevention and dispersal of pre-existing biofilms by crude RAU2C alginate lyase, highlighting its potential as an anti-biofilm agent against P. aeruginosa. The study highlights the efficacy of RAU2C alginate lyase as an anti-biofilm agent in controlling P. aeruginosa biofilms.</p>","PeriodicalId":8929,"journal":{"name":"Biotechnology Letters","volume":"47 1","pages":"9"},"PeriodicalIF":2.0,"publicationDate":"2024-11-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142765743","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":"Coupling ultrasound and membrane filtration for the fractionation of Spirulina platensis sp. and the recovery of phycocyanin and pigment-free proteins.","authors":"Sara Obeid, Hussein Rida, Jérôme Peydecastaing, Hosni Takache, Ali Ismail, Pierre-Yves Pontalier","doi":"10.1007/s10529-024-03541-9","DOIUrl":"10.1007/s10529-024-03541-9","url":null,"abstract":"<p><p>The cyanobacterium Spirulina platensis was subjected to a fractionation process involving ultrasound-assisted extraction and membrane filtration to obtain a pure phycocyanin fraction and a clarified colorless protein fraction free of chlorophyll and carotenoids. The effects of pressure and power on total protein release were assessed. The retention of the extracted proteins was then assessed by ultrafiltration, with and without ammonium sulfate precipitation. Total protein recovery yields reached 97% in aqueous solution, at a low frequency (12 kHz), atmospheric pressure, and with an ultrasonic power of 200 Watts (W). Ammonium sulfate (25% w/v) precipitation was used to remove pigments and impurities from the crude protein extract. Finally, semi-frontal ultrafiltration resulted in high levels of C-phycocyanin recovery in the retentate: 95% and 91% with 10 and 100 kDa-cutoff membranes, respectively. However, the levels of total non-pigmented proteins in the permeate compartment did not exceed 67% with a 100 kDa-cutoff membrane. A fractionation process is proposed here for the valorization of two different protein fractions from Spirulina platensis.</p>","PeriodicalId":8929,"journal":{"name":"Biotechnology Letters","volume":"47 1","pages":"8"},"PeriodicalIF":2.0,"publicationDate":"2024-11-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11607054/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142754459","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":"The influence of ZIF-L in a microbial fuel cell (MFC) cathode for oxygen reduction reaction (ORR).","authors":"Müşerref Eryılmaz, Janset Otuzoğlu, Ulas Tezel, Oktay Demircan","doi":"10.1007/s10529-024-03548-2","DOIUrl":"10.1007/s10529-024-03548-2","url":null,"abstract":"<p><p>Microbial fuel cells (MFCs) utilize the metabolic activities of microorganisms, through which the chemical energy is directly converted into electrical energy. Bacteria produce electrons by means of oxidation of organic/inorganic substrates within the MFCs. Metal organic frameworks (MOFs) that are porous coordination polymers have gained much interest in the field of efficient catalysts due to their unique characteristics. The utilization of MOF catalysts for oxygen reduction reaction (ORR) in the MFC cathode is one of the most remarkable research areas in material science. MOF (zeolitic imidazole framework-leaf like, ZIF-L) decorated cathode system was employed for the first time in MFC to monitor the improvement in performance by taking advantages of both electrocatalytic activity and porosity of MOFs for the utilization of bioelectrons for ORR. Analysis of ORR performance of ZIF-L/carbon black (CB) composite cathode demonstrated that ZIF-L containing cathode system had an improved ORR activity compared to MFC cathode materials in the literature. The remarkable current density value of 2.1 mA cm^-2 and the maximum power density value of 1,462 mW m^-2 at room temperature revealed that ZIF-L decorated cathode is an excellent alternative for efficient reduction of oxygen in MFCs.</p>","PeriodicalId":8929,"journal":{"name":"Biotechnology Letters","volume":"47 1","pages":"5"},"PeriodicalIF":2.0,"publicationDate":"2024-11-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142749838","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":"Synergistic anticancer effects of interleukin-21 combined with therapeutic peptides in multiple cancer cells.","authors":"Muhammad Akram, Nao Akusa Fujimura, Saad Tahir, Rabia Abbas, Mohsin Ahmad Khan, Kausar Malik, Nadeem Ahmed","doi":"10.1007/s10529-024-03544-6","DOIUrl":"10.1007/s10529-024-03544-6","url":null,"abstract":"<p><strong>Background: </strong>Interleukin-21 (IL-21) is a cytokine produced by various cell types, including T cells, natural killer cells, myeloid cells, and B cells, and has a broad range of potential applications in cancer therapy. To improve the therapeutic index, we explored the use of fusion technologies that involved linking other anticancer peptides to the IL-21 gene using specific linkers.</p><p><strong>Objectives: </strong>This study aimed to compare the anticancer potential of IL-21 and IL-21 fusion proteins.</p><p><strong>Methods: </strong>Antimicrobial peptides possessing anticancer properties were fused with IL-21 gene using a flexible linker (-GGGGS-), and the resulting construct was inserted into the pSecTag2a mammalian expression vector. The cassette was transfected into several cancer cell lines including H1 HeLa, HepG2, MCF-7, MDA-MB-231, HCT-116, HCC-1954, HEK-293, and SF-767. The cytotoxic effects of IL-21 and fusion proteins were evaluated using MTT, Caspase-3, LDH, and scratch assays.</p><p><strong>Results: </strong>The IL-21-Tachyplesin I fusion protein had the strongest antiproliferative activity against all tested cancer cells, followed by IL21-LPSBD2 and IL-21. In contrast, IL21-Cop A3, IL21-CSP I-Plus, and IL21-RGD Temporin-Las did not inhibit the viability of cancer cells.</p><p><strong>Conclusion: </strong>Fusion technology is a promising therapeutic technique that can be used to enhance the cytotoxicity and antiproliferative activity of anticancer proteins such as IL-21.</p>","PeriodicalId":8929,"journal":{"name":"Biotechnology Letters","volume":"47 1","pages":"7"},"PeriodicalIF":2.0,"publicationDate":"2024-11-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142749784","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":"Functional decoration of elastin-like polypeptides-based nanoparticles with a modular assembly via isopeptide bond formation.","authors":"Jun Yamaguchi, Kei Nishida, Eiry Kobatake, Masayasu Mie","doi":"10.1007/s10529-024-03549-1","DOIUrl":"10.1007/s10529-024-03549-1","url":null,"abstract":"<p><p>Temperature-responsive elastin-like polypeptides (ELPs) exhibit a low critical solution temperature-type phase transition and offer potential as useful materials for the construction of nanoparticles. Herein, we developed a novel decoration method for ELP-based nanoparticles via isopeptide bond formation with the SnoopTag/SnoopCatcher system that is not affected by the heating process required for particle formation. A mixture of a fusion protein of ELP and poly(aspartic acid) (poly(D)), known as ELP-poly(D), and ELP-poly(D) fused with SnoopCatcher (ELP-poly(D)-SnC) formed protein nanoparticles as a result of the temperature responsiveness of ELP, with the resultant nanoparticles displaying the SnoopCatcher binding domain on their surfaces. In the present study, two model proteins fused to SnoopTag were displayed on the surfaces of protein nanoparticles constructed from ELP-poly(D)-SnC and ELP-poly(D). The model proteins are enhanced green fluorescent protein (EGFP) and Renilla luciferace (Rluc), which exhibits luminescent capability and weak thermostability, respectively. EGFP on the particle surface was found to retain 48.7% activity, while Rluc exhibited almost full activity, as calculated from the binding efficiency and nanoparticle activities recovered after purification. ELP-based nanoparticles containing the SnoopTag/SnoopCatcher system offer the opportunity for particle decoration with a wide range of functional proteins via isopeptide bond formation.</p>","PeriodicalId":8929,"journal":{"name":"Biotechnology Letters","volume":"47 1","pages":"6"},"PeriodicalIF":2.0,"publicationDate":"2024-11-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142749783","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":"Rolling circle amplification cooperating crRNA switch for direct and sensitive methicillin-resistant Staphylococcus aureus (MRSA) analysis.","authors":"Junling Qiu, Chang Liu, Yuxia Zhu","doi":"10.1007/s10529-024-03550-8","DOIUrl":"10.1007/s10529-024-03550-8","url":null,"abstract":"<p><p>Evaluating the methicillin resistance of Staphylococcus aureus (S. aureus) is highly important for adapting nursing strategies. Nevertheless, the identification of methicillin-resistant S. aureus (MRSA) that is both sensitive and reliable continues to pose a significant obstacle. This study describes a method for detecting MRSA using a combination of fixed rolling circle amplification (RCA) and the exonuclease-iii (Exo-iii) assisted CRISPR-Cas12a system for signal amplification. When MRSA is present, the interaction between the \"b\" chain in the capture probe and MRSA allows the \"a\" chain to be exposed. This \"a\" chain acts as a primer to initiate the fixed RCA process. The H probe, which includes the crRNA segment, forms a bond with the RCA product and then releases the crRNA segment with the aid of Exo-iii. The Cas12a protein, when combined with the crRNA, generates an activated CRISPR-Cas12a system that cleaves the \"Reporter\" probe, resulting in the production of fluorescent signals. Furthermore, this fluorescent test has been utilized for the examination of clinical samples with a satisfactory rate of retrieval. Based on the elegant design, the proposed method exhibited a low detection limit of 4.6 cfu/mL, while maintaining a high specificity for MRSA even from a mixture of several interfering bacteria. Due to its cost-effectiveness, simplicity, and adaptability, the sensing system shows potential as a platform for detecting MRSA and evaluating postoperative nursing for stomach cancer patients.</p>","PeriodicalId":8929,"journal":{"name":"Biotechnology Letters","volume":"47 1","pages":"4"},"PeriodicalIF":2.0,"publicationDate":"2024-11-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142708971","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":"Butea monosperma bark extract: a natural boost for osteogenesis via Wnt/β-catenin pathway activation in adipose-derived mesenchymal stem cells.","authors":"Rebu Sundar, Gayathri Sundar, Annie John, Annie Abraham","doi":"10.1007/s10529-024-03545-5","DOIUrl":"10.1007/s10529-024-03545-5","url":null,"abstract":"<p><strong>Purpose: </strong>To investigate the impact of Butea monosperma (BM) bark extract on the osteogenic differentiation potential of rat adipose-derived mesenchymal stem cells (rADMSCs) and to elucidate the involvement of Wnt/β-catenin pathway in mediating this osseous effect.</p><p><strong>Methods: </strong>Characterizations (antioxidant assays, FTIR and LC/MS analyses) and docking studies (in silico) were performed to evaluate the presence of phytochemicals in the BM extract and their binding capacity to that of the frizzled receptor. rADMSCs were isolated and characterised for its differentiation potential of osteogenesis for stemness. Dose fixation, cytotoxicity, osteogenic differentiation (calcium, mineral deposition, alkaline phosphatase and osteocalcin) and gene expression (osteocalcin, Col1, osteonectin, Bmp2, Runx2, Wnt2, and β-catenin-14 and 28 days) of the extract were also evaluated in vitro.</p><p><strong>Results: </strong>FTIR and LC/MS analyses unveiled the phytochemicals in the extract and with docking studies confirmed their interaction with the frizzled receptor of Wnt/β-catenin pathway. rADMSCs were isolated and differentiated in the presence of the osteogenic induction medium. Dose fixation studies, cytotoxicity and cell viability assessments demonstrated the phytochemicals concentration-dependent cytotoxicity. The presence of specific bone markers highlighted the osteogenic differentiation potential of the phytochemicals. Furthermore, gene expression studies of rADMSCs depicted a heightened bone-forming capacity potentially facilitated by the activation of Wnt/β-catenin pathway.</p><p><strong>Conclusion: </strong> The phytochemicals of BM promoted the osteogenic differentiation of rADMSCs through the activation of the signalling Wnt/β-Catenin pathway, as evidenced by the significant upregulation of early and late bone markers. The phytochemicals may therefore be positioned as promising therapeutic agents for enhancing bone regeneration, offering new avenues for regenerative medicine.</p>","PeriodicalId":8929,"journal":{"name":"Biotechnology Letters","volume":"47 1","pages":"3"},"PeriodicalIF":2.0,"publicationDate":"2024-11-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142708964","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":"Computer-aided rational design of a mRNA vaccine against Guanarito mammarenavirus.","authors":"Mohibullah Shah, Asifa Sarfraz, Muhammad Shehroz, Asia Perveen, Samavia Jaan, Aqal Zaman, Umar Nishan, Arlindo A Moura, Riaz Ullah, Zafar Iqbal, Mohamed A Ibrahim","doi":"10.1007/s10529-024-03543-7","DOIUrl":"10.1007/s10529-024-03543-7","url":null,"abstract":"<p><strong>Purpose: </strong>Guanarito mammarenavirus (GTOV) is a highly pathogenic virus that leads to Venezuelan hemorrhagic fever (VHF). Despite being a severe disease, there are currently no commercially available drugs or vaccines for its prevention.</p><p><strong>Methods: </strong>Here we computationally formulated a mRNA vaccine construct (VC) from the genome of GTOV to produce immunity against its infections. Two proteins, namely zinc-finger motif protein (NP_899220.1), and nucleocapsid protein (NP_899211.1) were screened as potential candidates for downstream analysis.</p><p><strong>Results: </strong>We determined the T and B cell epitopes of the candidate proteins. The resulting epitopes were analyzed, and the best epitopes were utilized in the formation of the peptide vaccine construct. The secondary and tertiary structures of the peptide construct were predicted and validated. Docking was conducted to check the binding energy of the designed peptide vaccine with the human immune receptors, namely TLR2 and TLR4. Our designed vaccine showed stable interactions with the HLA molecules, as verified through normal mode and MD simulation analysis. The immune simulation results indicated a positive immune response against the construct. A potentially stable mRNA vaccine was formulated by adding of sequences such as the Kozak, Goblin 5' UTR, tPA-signal peptide, MITD, 3' UTRs, and a poly(A) tail to the peptide vaccine construct. Lastly, the expression probability of the mRNA vaccine was confirmed in the expression system of E. coli strain K12.</p><p><strong>Conclusion: </strong>The designed vaccine showed the potential to elicit an immune response against the GTOV infection; however, experimental validation is recommended to verify the in-silico findings of this study.</p>","PeriodicalId":8929,"journal":{"name":"Biotechnology Letters","volume":"47 1","pages":"2"},"PeriodicalIF":2.0,"publicationDate":"2024-11-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142708966","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":"Production of derivatives of α-terpineol by bacterial CYP102A1 enzymes.","authors":"Jeong-Hoon Kim, Chan Mi Park, Hae Chan Jeong, Sungbeom Lee, Chul-Ho Yun","doi":"10.1007/s10529-024-03540-w","DOIUrl":"10.1007/s10529-024-03540-w","url":null,"abstract":"<p><p>The monooxygenase activity of engineered CYP102A1 on α-terpineol was investigated. CYP102A1 M850 mutant (F11Y/R47L/D68G/F81I/F87V/E143G/L188Q/E267V/H408R) showed the highest catalytic activity toward α-terpineol among the engineered mutants produced by random mutagenesis. The major product (P1) of α-terpineol, p-menth-1-ene-3,8-diol, was characterized by high-performance liquid chromatography, gas-chromatography mass spectrometry, and nuclear magnetic resonance spectroscopy. Three minor products (P2-P4) of α-terpineol were considered as 6-hydroxy-α,α,4-trimethyl-3-cyclohexene-1-methanol (P2), trans-sobrerol (P3), and carvone hydrate (P4). Optimal conditions for product formation were determined as pH 7.0 and 30 °C. Production of p-menth-1-ene-3,8-diol was 0.87 mM at 1 h. Structure modeling using PyMOL and CAVER Web 1.2 server indicated that several mutations of CYP102A1 M850 were involved in access tunnels and active sites, resulting in increased activity toward α-terpineol. The major product, p-menth-1-ene-3,8-diol, of α-terpineol was produced by engineered CYP102A1 M850 via regioselective carbon hydroxylation. The engineered CYP102A1 could be a suitable biocatalyst for producing α-terpineol derivatives.</p>","PeriodicalId":8929,"journal":{"name":"Biotechnology Letters","volume":"47 1","pages":"1"},"PeriodicalIF":2.0,"publicationDate":"2024-11-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142708970","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}