Biotechnology Letters最新文献_第5页

Factors affecting rAAV titers during triple-plasmid transient transfection in HEK-293 cells 影响 HEK-293 细胞中三重质粒瞬时转染过程中 rAAV 滴度的因素

IF 2.7 4区生物学

Biotechnology Letters Pub Date : 2024-09-11 DOI: 10.1007/s10529-024-03520-0

Martina Pistek, Peter Andorfer, Reingard Grabherr, Barbara Kraus, Juan A. Hernandez Bort

{"title":"Factors affecting rAAV titers during triple-plasmid transient transfection in HEK-293 cells","authors":"Martina Pistek, Peter Andorfer, Reingard Grabherr, Barbara Kraus, Juan A. Hernandez Bort","doi":"10.1007/s10529-024-03520-0","DOIUrl":"https://doi.org/10.1007/s10529-024-03520-0","url":null,"abstract":"The efficiency of triple-plasmid transfection in recombinant Adeno-Associated Virus (rAAV) production was analyzed by examining two distinct HEK-293 cells lines. These were categorized as high producer (HP) and low producer (LP) based on their differing levels of productivity under identical conditions. Analysis of RNA expression levels of viral genes revealed disparities in plasmid derived gene expression between the cell lines. Further assessment of transfection efficiency utilizing labeled plasmids revealed lower plasmid uptake and less efficient nuclear transport in LP cell line. Additionally, we observed inferior translation activity in LP, contributing to its shortcomings in overall productivity. In our attempt to optimize plasmid ratios to enhance fully packaged rAAV particle yield, we discovered cell-line-specific optimization potential. The findings highlight the transfection's complexity, urging tailored strategies for improved rAAV production based on each cell line's characteristics, enhancing understanding and guiding further efficiency optimization in rAAV production.","PeriodicalId":8929,"journal":{"name":"Biotechnology Letters","volume":"3 1","pages":""},"PeriodicalIF":2.7,"publicationDate":"2024-09-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142186272","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Succession from acetoclastic to hydrogenotrophic microbial community during sewage sludge anaerobic digestion for bioenergy production 用于生物能源生产的污水污泥厌氧消化过程中从嗜醋微生物群落到嗜氢微生物群落的演替

IF 2.7 4区生物学

Biotechnology Letters Pub Date : 2024-09-11 DOI: 10.1007/s10529-024-03528-6

Mariana Erthal Rocha, Norberto Mangiavacchi, Marcia Marques, Lia Teixeira

{"title":"Succession from acetoclastic to hydrogenotrophic microbial community during sewage sludge anaerobic digestion for bioenergy production","authors":"Mariana Erthal Rocha, Norberto Mangiavacchi, Marcia Marques, Lia Teixeira","doi":"10.1007/s10529-024-03528-6","DOIUrl":"https://doi.org/10.1007/s10529-024-03528-6","url":null,"abstract":"To assess microbial dynamics during anaerobic digestion (AD) of sewage sludge (SWS) from a municipal Wastewater Treatment Plant (WWTP), a Biochemical Methane Potential (BMP) assay at 37 °C under mono-digestion conditions was conducted. Utilizing the Illumina MiSeq platform, 16S ribosomal RNA (rRNA) gene sequencing unveiled a core bacterial community in the solid material, showcasing notable variations in profiles. The research investigates changes in microbial communities and metabolic pathways to understand their impact on the efficiency of the digestion process. Prior to AD, the relative abundance in SWS was as follows: Proteobacteria > Bacteroidota > Actinobacteriota. Post-AD, the relative abundance shifted to Firmicutes > Synergistota > Proteobacteria, with Sporanaerobacter and Clostridium emerging as dominant genera. Notably, the methanogenic community underwent a metabolic pathway shift from acetoclastic to hydrogenotrophic in the lab-scale reactors. At the genus level, Methanosaeta, Methanolinea, and Methanofastidiosum predominated initially, while post-AD, Methanobacterium, Methanosaeta, and Methanospirillum took precedence. This metabolic transition may be linked to the increased abundance of Firmicutes, particularly Clostridia, which harbor acetate-oxidizing bacteria facilitating the conversion of acetate to hydrogen.","PeriodicalId":8929,"journal":{"name":"Biotechnology Letters","volume":"60 1","pages":""},"PeriodicalIF":2.7,"publicationDate":"2024-09-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142224326","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Research on the application of heterotrophic nitrification-aerobic denitrification bacteria in membrane bioreactor (MBR) 异养硝化-好氧反硝化菌在膜生物反应器（MBR）中的应用研究

IF 2.7 4区生物学

Biotechnology Letters Pub Date : 2024-09-11 DOI: 10.1007/s10529-024-03529-5

Tianrui Zhai, Tiantao Zhao, Yuhao Zhong, Peipei Chen, Guojian Li, Liang Teng, Lijie Zhang, Hao Liu

引用次数: 0

The repertoire and levels of secondary metabolites in microbial cocultures depend on the inoculation ratio: a case study involving Aspergillus terreus and Streptomyces rimosus. 微生物共培养物中次生代谢物的种类和水平取决于接种比例：一项涉及赤曲霉和链霉菌的案例研究。

IF 2 4区生物学

Biotechnology Letters Pub Date : 2024-08-01 Epub Date: 2024-06-06 DOI: 10.1007/s10529-024-03500-4

Tomasz Boruta, Grzegorz Englart, Martyna Foryś, Weronika Pawlikowska

{"title":"The repertoire and levels of secondary metabolites in microbial cocultures depend on the inoculation ratio: a case study involving Aspergillus terreus and Streptomyces rimosus.","authors":"Tomasz Boruta, Grzegorz Englart, Martyna Foryś, Weronika Pawlikowska","doi":"10.1007/s10529-024-03500-4","DOIUrl":"10.1007/s10529-024-03500-4","url":null,"abstract":"Objective: The aim of this study was to determine the influence of the inoculation volume ratio on the production of secondary metabolites in submerged cocultures of Aspergillus terreus and Streptomyces rimosus.Results: The shake flask cocultures were initiated by using 23 inoculum variants that included different volumes of A. terreus and S. rimosus precultures. In addition, the axenic controls were propagated in parallel with the cocultures. UPLC‒MS analysis revealed the presence of 15 secondary metabolites, 12 of which were found both in the \"A. terreus vs. S. rimosus\" cocultures and axenic cultures of either A. terreus or S. rimosus. The production of the remaining 3 molecules was recorded solely in the cocultures. The repertoire and quantity of secondary metabolites were evidently dependent on the inoculation ratio. It was also noted that detecting filamentous structures resembling typical morphological forms of a given species was insufficient to predict the presence of a given metabolite.Conclusions: The modification of the inoculation ratio is an effective strategy for awakening and enhancing the production of secondary metabolites that are not biosynthesized under axenic conditions.","PeriodicalId":8929,"journal":{"name":"Biotechnology Letters","volume":" ","pages":"601-614"},"PeriodicalIF":2.0,"publicationDate":"2024-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11217084/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141282895","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}

引用次数: 0

Mutational analysis of pig tissue factor pathway inhibitor α to increase anti-coagulation activity in pig-to-human xenotransplantation. 对猪组织因子通路抑制剂α进行突变分析，以提高猪-人异种移植中的抗凝血活性。

IF 2 4区生物学

Biotechnology Letters Pub Date : 2024-08-01 Epub Date: 2024-06-13 DOI: 10.1007/s10529-024-03505-z

Chang-Hee Lee, Hyeon Jeong Lee, Si-Won Park, Jiyoon Shin, Seok-Jin Kang, In-Byung Park, Hyun Kyung Kim, Taehoon Chun

{"title":"Mutational analysis of pig tissue factor pathway inhibitor α to increase anti-coagulation activity in pig-to-human xenotransplantation.","authors":"Chang-Hee Lee, Hyeon Jeong Lee, Si-Won Park, Jiyoon Shin, Seok-Jin Kang, In-Byung Park, Hyun Kyung Kim, Taehoon Chun","doi":"10.1007/s10529-024-03505-z","DOIUrl":"10.1007/s10529-024-03505-z","url":null,"abstract":"Blood coagulation mediated by pig tissue factor (TF), which is expressed in pig tissues, causes an instant blood-mediated inflammatory reaction during pig-to-human xenotransplantation. Previously, we generated a soluble pig tissue factor pathway inhibitor α fusion immunoglobulin (TFPI-Ig) which inhibits pig TF activity more efficiently than human TFPI-Ig in human plasma. In this study, we generated several pig TFPI-Ig mutants and tested the efficacy of these mutants in preventing pig-to-human xenogeneic blood coagulation. Structurally important amino acid residues of pig TFPI-Ig were changed into different residues by site-directed mutagenesis. Subsequently, a retroviral vector encoding each cDNA of several pig TFPI-Ig mutants was cloned and transduced into CHO-K1 cells. After establishing stable cell lines expressing each of the pig TFPI-Ig mutants, soluble proteins were produced and purified for evaluating their inhibitory effects on pig TF-mediated blood coagulation in human plasma. The replacement of K36 and K257 with R36 and H257, respectively, in pig TFPI-Ig more efficiently blocked pig TF activity in human plasma when compared with the wild-type pig TFPI-Ig. These results may provide additional information to understand the structure of pig TFPIα, and an improved pig TFPI-Ig variant that more efficiently blocks pig TF-mediated blood coagulation during pig-to-human xenotransplantation.","PeriodicalId":8929,"journal":{"name":"Biotechnology Letters","volume":" ","pages":"521-530"},"PeriodicalIF":2.0,"publicationDate":"2024-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141316697","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Diffusion-driven fed-batch fermentation in perforated ring flasks. 穿孔环形烧瓶中的扩散驱动饲料批量发酵。

IF 2 4区生物学

Biotechnology Letters Pub Date : 2024-08-01 Epub Date: 2024-05-17 DOI: 10.1007/s10529-024-03493-0

Clara Lüchtrath, Felix Lamping, Sven Hansen, Maurice Finger, Jørgen Magnus, Jochen Büchs

{"title":"Diffusion-driven fed-batch fermentation in perforated ring flasks.","authors":"Clara Lüchtrath, Felix Lamping, Sven Hansen, Maurice Finger, Jørgen Magnus, Jochen Büchs","doi":"10.1007/s10529-024-03493-0","DOIUrl":"10.1007/s10529-024-03493-0","url":null,"abstract":"Purpose: Simultaneous membrane-based feeding and monitoring of the oxygen transfer rate shall be introduced to the newly established perforated ring flask, which consists of a cylindrical glass flask with an additional perforated inner glass ring, for rapid bioprocess development.Methods: A 3D-printed adapter was constructed to enable monitoring of the oxygen transfer rate in the perforated ring flasks. Escherichia coli experiments in batch were performed to validate the adapter. Fed-batch experiments with different diffusion rates and feed solutions were performed.Results: The adapter and the performed experiments allowed a direct comparison of the perforated ring flasks with Erlenmeyer flasks. In batch cultivations, maximum oxygen transfer capacities of 80 mmol L-1 h-1 were reached with perforated ring flasks, corresponding to a 3.5 times higher capacity than in Erlenmeyer flasks. Fed-batch experiments with a feed reservoir concentration of 500 g glucose L-1 were successfully conducted. Based on the oxygen transfer rate, an ammonium limitation could be observed. By adding 40 g ammonium sulfate L-1 to the feed reservoir, the limitation could be prevented.Conclusion: The membrane-based feeding, an online monitoring technique, and the perforated ring flask were successfully combined and offer a new and promising tool for screening and process development in biotechnology.","PeriodicalId":8929,"journal":{"name":"Biotechnology Letters","volume":" ","pages":"571-582"},"PeriodicalIF":2.0,"publicationDate":"2024-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11217090/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140955731","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}

引用次数: 0

Engineering of itaconic acid pathway via co-localization of CadA and AcnA in recombinant Escherichia coli. 通过 CadA 和 AcnA 在重组大肠杆菌中的共定位改造衣康酸途径。

IF 2 4区生物学

Biotechnology Letters Pub Date : 2024-08-01 Epub Date: 2024-05-29 DOI: 10.1007/s10529-024-03496-x

Kim-Ngan T Tran, Jaehoon Jeong, Soon Ho Hong

引用次数: 0

Isolation of a facultative methanotroph Methylocystis iwaonis SD4 from rice rhizosphere and establishment of rapid genetic tools for it. 从水稻根瘤中分离出一种兼性甲烷营养体 Methylocystis iwaonis SD4，并为其建立快速遗传工具。

IF 2 4区生物学

Biotechnology Letters Pub Date : 2024-08-01 Epub Date: 2024-05-11 DOI: 10.1007/s10529-024-03495-y

Yinghui Wang, Yuying Wang, Keyu Zhou, Haili Zhang, Minggen Cheng, Baozhan Wang, Xin Yan

{"title":"Isolation of a facultative methanotroph Methylocystis iwaonis SD4 from rice rhizosphere and establishment of rapid genetic tools for it.","authors":"Yinghui Wang, Yuying Wang, Keyu Zhou, Haili Zhang, Minggen Cheng, Baozhan Wang, Xin Yan","doi":"10.1007/s10529-024-03495-y","DOIUrl":"10.1007/s10529-024-03495-y","url":null,"abstract":"Methanotrophs of the genus Methylocystis are frequently found in rice paddies. Although more than ten facultative methanotrophs have been reported since 2005, none of these strains was isolated from paddy soil. Here, a facultative methane-oxidizing bacterium, Methylocystis iwaonis SD4, was isolated and characterized from rhizosphere samples of rice plants in Nanjing, China. This strain grew well on methane or methanol but was able to grow slowly using acetate or ethanol. Moreover, strain SD4 showed sustained growth at low concentrations of methane (100 and 500 ppmv). M. iwaonis SD4 could utilize diverse nitrogen sources, including nitrate, urea, ammonium as well as dinitrogen. Strain SD4 possessed genes encoding both the particulate methane monooxygenase and the soluble methane monooxygenase. Simple and rapid genetic manipulation methods were established for this strain, enabling vector transformation and unmarked genetic manipulation. Fast growth rate and efficient genetic tools make M. iwaonis SD4 an ideal model to study facultative methanotrophs, and the ability to grow on low concentration of methane implies its potential in methane removal.","PeriodicalId":8929,"journal":{"name":"Biotechnology Letters","volume":" ","pages":"713-724"},"PeriodicalIF":2.0,"publicationDate":"2024-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140908047","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Optimized detection of Salmonella typhimurium using aptamer lateral flow assay. 利用aptamer横向流动检测法优化鼠伤寒沙门氏菌的检测。

IF 2 4区生物学

Biotechnology Letters Pub Date : 2024-08-01 Epub Date: 2024-05-28 DOI: 10.1007/s10529-024-03484-1

Nafise Abedi, Mehdi Zeinoddini, Mohammad Shoushtari

{"title":"Optimized detection of Salmonella typhimurium using aptamer lateral flow assay.","authors":"Nafise Abedi, Mehdi Zeinoddini, Mohammad Shoushtari","doi":"10.1007/s10529-024-03484-1","DOIUrl":"10.1007/s10529-024-03484-1","url":null,"abstract":"Salmonella typhimurium, a pathogenic bacterium with significant implications in medicine and the food industry, poses a substantial threat by causing foodborne illnesses such as typhoid fever. Accurate diagnosis of S. typhimurium is challenging due to its overlap symptoms with various diseases. This underscores the need for a precise and efficient diagnostic approach. In this study, we developed a biosensor using the Taguchi optimization method based on aptamer lateral flow assay (LFA) for the detection of S. typhimurium. Therefore, signal probe and nanobioprobe were designed using anti-Salmonella aptamer, conjugated with gold nanoparticles (GNPs), and used in LFA. The strategy of this test is based on a competitive format between the bacteria immobilized on the membrane and the bacteria present in the tested sample. Moreovere, the optimization of various factors affecting the aptamer LFA, including the concentration of bacteria (immobilized and into the sample) and the concentration of nanobioprop, were performed using the Taguchi test designing method. The data showed that the optimal conditions for the LFA reaction was 108 CFU/mL of immobilized bacteria and 1.5 μg/μL of nanobioprop concentration. Then, the visual detection limit of S. typhimurium was estimated as 105 CFU/mL. The reaction results were obtained within 20 min, and there were no significant cross-reactions with other food pathogens. In conclusion, the aptamer-LFA diagnostic method, optimized using the Taguchi approach, emerges as a reliable, straightforward, and accurate tool for the detection of S. typhimurium. Overall, this method can be a portable diagnostic kit for the detection and identification of bacteria.","PeriodicalId":8929,"journal":{"name":"Biotechnology Letters","volume":" ","pages":"583-592"},"PeriodicalIF":2.0,"publicationDate":"2024-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141160106","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Paddy straw saccharification using immobilized laccase on magnetized multiwall carbon nanotubes. 利用磁化多壁碳纳米管上的固定化漆酶对稻草进行糖化。

IF 2 4区生物学

Biotechnology Letters Pub Date : 2024-08-01 Epub Date: 2024-05-15 DOI: 10.1007/s10529-024-03494-z

Hasnol Azahari Natasha Yasmin, Balakrishnan Kunasundari, Siew Hoong Shuit, Mohamad Fahrurrazi Tompang

{"title":"Paddy straw saccharification using immobilized laccase on magnetized multiwall carbon nanotubes.","authors":"Hasnol Azahari Natasha Yasmin, Balakrishnan Kunasundari, Siew Hoong Shuit, Mohamad Fahrurrazi Tompang","doi":"10.1007/s10529-024-03494-z","DOIUrl":"10.1007/s10529-024-03494-z","url":null,"abstract":"The effective recovery of the immobilized enzymes using magnetic carriers has led to growing interest in this technology. The objective of this research was to evaluate the efficiency of immobilized laccase on magnetized multiwall carbon nanotubes (m-MWCNTs) in terms of stability and reusability. Laccases were efficiently adsorbed onto magnetized multiwall carbon nanotubes (m-MWCNTs) synthesized using water. The concentration of 7 mg laccase/mL was found to be ideal for immobilization. The optimal activity of both free and immobilized laccases was observed at pH 5, while for the latter, the optimal temperature was shifted from 40 to 50 °C. Compared to the free laccase, the immobilized laccase exhibited a greater range of stability at more extreme temperatures. At the fourth cycle of reactions, the immobilized laccase exhibited more than 60% relative activity in terms of reusability. Based on the fourier-transform infrared spectroscopy (FTIR) peak at 2921 cm-1, saccharification of paddy straw using immobilized laccase verified lignin degradation. The easy recovery of the immobilized laccase on m-MWCNTs lends credence to its potential use in biomass hydrolysis.","PeriodicalId":8929,"journal":{"name":"Biotechnology Letters","volume":" ","pages":"559-569"},"PeriodicalIF":2.0,"publicationDate":"2024-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140921166","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0