Journal of biotechnology最新文献_第10页

3D printed autoclavable biocompatible biodegradable bioreactor vessels with integrated sparger made from poly-lactic acid 三维打印的可高压灭菌的生物相容性生物可降解生物反应器容器，带有聚乳酸制成的集成喷射器。

IF 4.1 2区生物学

Journal of biotechnology Pub Date : 2024-06-04 DOI: 10.1016/j.jbiotec.2024.06.001

Lena Achleitner , Martina Winter , Peter Satzer

{"title":"3D printed autoclavable biocompatible biodegradable bioreactor vessels with integrated sparger made from poly-lactic acid","authors":"Lena Achleitner , Martina Winter , Peter Satzer","doi":"10.1016/j.jbiotec.2024.06.001","DOIUrl":"10.1016/j.jbiotec.2024.06.001","url":null,"abstract":"<div><p>3D printing has become widespread for the manufacture of parts in various industries and enabled radically new designs. This trend has not spread to bioprocess development yet, due to a lack of material suitable for the current workflow, including sterilization by autoclaving. This work demonstrates that commercially available heat temperature stable poly-lactic acid (PLA) can be used to easily manufacture novel bioreactor vessels with included features like harvest tubes and 3D printed spargers. Temperature responsiveness was tested for PLA, temperature stable PLA (PLA-HP) and glass for temperatures relevant for insect and mammalian cell culture, including temperature shifts within the process. Stability at 27 °C and 37 °C as well as temperature shifts to 22 °C and 32 °C showed acceptable performance with slightly higher temperature overshoot for 3D printed vessels. A stable temperature is reached after 2 h for PLA, 3 h for PLA-HP and 1 h for glass reactors. Temperature can be maintained with a fluctuation of 0.1 °C for all materials. A 3D printed sparger design directly integrated into the vessel wall and bottom was tested under three different conditions (0.3 SLPH and 27 °C, 3 SLPH and 37 °C and 13 SLPH and 37 °C). The 3D printed sparger showed a better <em>k</em><sub><em>L</em></sub><em>a</em> than the L-Sparger with more pronounced differences for higher flowrates. An insect cell culture run in the novel vessel exhibited the same growth behavior as that in standard glass vessels, reaching the same maximum cell concentration. Being 3D printed from biodegradable materials, these bioreactors offer design flexibility for novel bioreactor formats. Additionally, their autoclavability allows seamless integration into standard workflows.</p></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":null,"pages":null},"PeriodicalIF":4.1,"publicationDate":"2024-06-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S0168165624001585/pdfft?md5=84eaa92ae8eab00368f5f0b282323f19&pid=1-s2.0-S0168165624001585-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141261903","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"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

Effective cultivation conditions and safety evaluation of filamentous cyanobacteria producing phycocyanins with antiglycation activities 产生具有抗糖化活性的藻蓝蛋白的丝状蓝藻的有效培养条件和安全性评价。

IF 4.1 2区生物学

Journal of biotechnology Pub Date : 2024-06-04 DOI: 10.1016/j.jbiotec.2024.06.003

Jinichi Aoki , Takato Ozaki , Runa Koshikawa , Daisaku Sasaki , Katsuyoshi Kitajima , Yuta Yoshida , Hiromi Nakajima , Munehiko Asayama

{"title":"Effective cultivation conditions and safety evaluation of filamentous cyanobacteria producing phycocyanins with antiglycation activities","authors":"Jinichi Aoki , Takato Ozaki , Runa Koshikawa , Daisaku Sasaki , Katsuyoshi Kitajima , Yuta Yoshida , Hiromi Nakajima , Munehiko Asayama","doi":"10.1016/j.jbiotec.2024.06.003","DOIUrl":"10.1016/j.jbiotec.2024.06.003","url":null,"abstract":"<div><p>We investigated suitable culture conditions for the production of the blue pigment phycocyanin (PC) from the unique filamentous cyanobacteria <em>Pseudanabaena</em> sp. ABRG5-3 and <em>Limnothrix</em> sp. SK1-2-1. White, green, or red LED irradiation at 30 μmol photons/m<sup>2</sup>/s was effective for phycocyanin production when compared with <em>Arthrospira platensis</em> (<em>Spirulina</em>) sp. NIES-39, which is generally grown under high light irradiation. To investigate the safety of the cyanobacteria, ABRG5-3 cells were subjected to Ames (reverse mutation) tests and single oral-dose rat studies, which revealed non-mutagenic and non-toxic properties. When three purified phycocyanins (abPC, skPC, and spPC) were subjected to agarose gel electrophoresis, they showed different mobility, indicating that each phycocyanin has unique properties. abPC exhibited strong antiglycation activities as novel function.</p></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":null,"pages":null},"PeriodicalIF":4.1,"publicationDate":"2024-06-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S0168165624001603/pdfft?md5=de40262f067118c2753b023e5cc21548&pid=1-s2.0-S0168165624001603-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141283787","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"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

Enhancement of production of glycoalkaloids by elicitors along with characterization of gene expression of pathways in Solanum xanthocarpum 通过诱导剂提高茄科植物糖生物碱的产量，并确定其通路基因表达的特征。

IF 4.1 2区生物学

Journal of biotechnology Pub Date : 2024-05-31 DOI: 10.1016/j.jbiotec.2024.05.008

Bharat Singh , Sheenu Nathawat , Anuja Saxena , Kiran Khangarot , Ram A. Sharma

{"title":"Enhancement of production of glycoalkaloids by elicitors along with characterization of gene expression of pathways in Solanum xanthocarpum","authors":"Bharat Singh , Sheenu Nathawat , Anuja Saxena , Kiran Khangarot , Ram A. Sharma","doi":"10.1016/j.jbiotec.2024.05.008","DOIUrl":"10.1016/j.jbiotec.2024.05.008","url":null,"abstract":"<div><p><em>Solanum xanthocarpum</em> fruits are used in the treatment of cough, fever, and heart disorders. It possesses antipyretic, hypotensive, antiasthmatic, aphrodisiac and antianaphylactic properties. In the present study, 24 elicitors (both biotic and abiotic) were used to enhance the production of glycoalkaloids in cell cultures of <em>S. xanthocarpum</em>. Four concentrations of elicitors were added into the MS culture medium. The maximum accumulation (5.56-fold higher than control) of demissidine was induced by sodium nitroprusside at 50 mM concentration whereas the highest growth of cell biomass (4.51-fold higher than control) stimulated by systemin at 30 mM concentration. A total of 17 genes of biosynthetic pathways of glycoalkaloids were characterized from the cells of <em>S. xanthocarpum</em>. The greater accumulation of demissidine was confirmed with the expression analysis of 11 key biosynthetic pathway enzymes e.g., acetoacetic-CoA thiolase, 3- hydroxy 3-methyl glutaryl synthase, β-hydroxy β-methylglutaryl CoA reductase, mevalonate kinase, farnesyl diphosphate synthase, squalene synthase, squalene epoxidase, squalene-2,3- epoxide cyclase, cycloartenol synthase, UDP-glucose: solanidine glucosyltransferase and UDP-rhamnose: solanidine rhamno-galactosyl transferase. The maximum expression levels of UDP-rhamnose: solanidine rhamno-galactosyl transferase gene was recorded in this study.</p></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":null,"pages":null},"PeriodicalIF":4.1,"publicationDate":"2024-05-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141199759","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"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

Investigating the influence of taurochenodeoxycholic acid (TCDCA) on pancreatic cancer cell behavior: An RNA sequencing approach 研究牛磺鹅去氧胆酸（TCDCA）对胰腺癌细胞行为的影响：一种 RNA 测序方法

IF 4.1 2区生物学

Journal of biotechnology Pub Date : 2024-05-28 DOI: 10.1016/j.jbiotec.2024.05.010

Eleonóra Gál , Shahram Parvaneh , Vanda Miklós , Péter Hegyi , Lajos Kemény , Zoltán Veréb , Viktória Venglovecz

{"title":"Investigating the influence of taurochenodeoxycholic acid (TCDCA) on pancreatic cancer cell behavior: An RNA sequencing approach","authors":"Eleonóra Gál , Shahram Parvaneh , Vanda Miklós , Péter Hegyi , Lajos Kemény , Zoltán Veréb , Viktória Venglovecz","doi":"10.1016/j.jbiotec.2024.05.010","DOIUrl":"10.1016/j.jbiotec.2024.05.010","url":null,"abstract":"<div><p>Pancreatic cancer (PC) poses a substantial global health challenge, ranking as the fourth leading cause of cancer-related deaths due to its high mortality rate. Late-stage diagnoses are common due to the absence of specific symptoms. Pancreatic ductal adenocarcinoma (PDAC) accounts for the majority of PC cases. Recent research has suggested a potential link between elevated serum levels of bile acids (BAs) and tumorigenesis of PDAC. This study aims to understand how taurochenodeoxycholic acid (TCDCA), a secondary BA, influences PDAC using RNA sequencing techniques on the Capan-1 cell line. We identified 2,950 differentially expressed genes (DEGs) following TCDCA treatment, with 1,597 upregulated and 1,353 downregulated genes. These DEGs were associated with critical PDAC pathways, including coagulation, angiogenesis, cell migration, and signaling regulation.</p><p>Furthermore, we reviewed relevant literature highlighting genes like DKK-1, KRT80, UPLA, and SerpinB2, known for their roles in PDAC tumorigenesis and metastasis. Our study sheds light on the complex relationship between BAs and PDAC, offering insights into potential diagnostic markers and therapeutic targets. Further research is needed to unravel these findings' precise mechanisms and clinical implications, potentially improving PDAC diagnosis and treatment.</p></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":null,"pages":null},"PeriodicalIF":4.1,"publicationDate":"2024-05-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S0168165624001500/pdfft?md5=36d602e1f6188b386644c2fe2ecda33f&pid=1-s2.0-S0168165624001500-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141179701","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"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

Heterologous expression of frog antimicrobial peptide Odorranain-C1 in Pichia pastoris: Biological characteristics and its application in food preservation 青蛙抗菌肽 Odorranain-C1 在 Pichia pastoris 中的异源表达：生物学特性及其在食品保鲜中的应用

IF 4.1 2区生物学

Journal of biotechnology Pub Date : 2024-05-23 DOI: 10.1016/j.jbiotec.2024.05.009

Mengru Li , Ruonan Zhou , Yuanyuan Wang , Yan Lu , Xinlei Chu , Chunming Dong

{"title":"Heterologous expression of frog antimicrobial peptide Odorranain-C1 in Pichia pastoris: Biological characteristics and its application in food preservation","authors":"Mengru Li , Ruonan Zhou , Yuanyuan Wang , Yan Lu , Xinlei Chu , Chunming Dong","doi":"10.1016/j.jbiotec.2024.05.009","DOIUrl":"https://doi.org/10.1016/j.jbiotec.2024.05.009","url":null,"abstract":"<div><p>To reduce food spoilage and deterioration caused by microbial contamination, antimicrobial peptides (AMPs) have gradually gained attention as a biological preservative. Odorranain-C1 is an α-helical cationic antimicrobial peptide extracted from the skin of frogs with broad-spectrum antimicrobial activity. In this study, we achieved the expression of Odorranain-C1 in <em>Pichia pastoris</em> (<em>P. pastoris</em>) (also known as <em>Komagataella phaffii</em>) by employing DNA recombination technology. The recombinant Odorranain-C1 showed broad-spectrum antibacterial activity and displayed a minimum inhibitory concentration within the range of 8–12 μg.mL<sup>-1</sup>. Meanwhile, Odorranain-C1 exhibited superior stability and lower hemolytic activity. Mechanistically, Odorranain-C1 disrupted the bacterial membrane's integrity, ultimately causing membrane rupture and subsequent cell death. In tilapia fillets preservation, Odorranain-C1 inhibited the total colony growth and pH variations, while also reducing the production of total volatile basic nitrogen (TVB-N) and thiobarbituric acid (TBA). In conclusion, these studies demonstrated the efficient recombinant expression of Odorranain-C1 in <em>P. pastoris</em>, highlighting its promising utilization in food preservation.</p></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":null,"pages":null},"PeriodicalIF":4.1,"publicationDate":"2024-05-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141090303","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"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

Up-regulation of Retrograde Response in yeast increases glycerol and reduces ethanol during wine fermentation 在葡萄酒发酵过程中，酵母逆行反应的上调会增加甘油，减少乙醇。

IF 4.1 2区生物学

Journal of biotechnology Pub Date : 2024-05-18 DOI: 10.1016/j.jbiotec.2024.05.007

Víctor Garrigós , Beatriz Vallejo , Esperanza Mollà-Martí , Cecilia Picazo , Emilien Peltier , Philippe Marullo , Emilia Matallana , Agustín Aranda

{"title":"Up-regulation of Retrograde Response in yeast increases glycerol and reduces ethanol during wine fermentation","authors":"Víctor Garrigós , Beatriz Vallejo , Esperanza Mollà-Martí , Cecilia Picazo , Emilien Peltier , Philippe Marullo , Emilia Matallana , Agustín Aranda","doi":"10.1016/j.jbiotec.2024.05.007","DOIUrl":"10.1016/j.jbiotec.2024.05.007","url":null,"abstract":"<div><p>Nutrient signaling pathways play a pivotal role in regulating the balance among metabolism, growth and stress response depending on the available food supply. They are key factors for the biotechnological success of the yeast <em>Saccharomyces cerevisiae</em> during food-producing fermentations. One such pathway is Retrograde Response, which controls the alpha-ketoglutarate supply required for the synthesis of amino acids like glutamate and lysine. Repressor <em>MKS1</em> is linked with the TORC1 complex and negatively regulates this pathway. Deleting <em>MKS1</em> from a variety of industrial strains causes glycerol to increase during winemaking, brewing and baking. This increase is accompanied by a reduction in ethanol production during grape juice fermentation in four commercial wine strains. Interestingly, this does not lead volatile acidity to increase because acetic acid levels actually lower. Aeration during winemaking usually increases acetic acid levels, but this effect reduces in the <em>MKS1</em> mutant. Despite the improvement in the metabolites of oenological interest, it comes at a cost given that the mutant shows slower fermentation kinetics when grown in grape juice, malt and laboratory media and using glucose, sucrose and maltose as carbon sources. The deletion of <em>RTG2</em>, an activator of Retrograde Response that acts as an antagonist of <em>MKS1</em>, also results in a defect in wine fermentation speed. These findings suggest that the deregulation of this pathway causes a fitness defect. Therefore, manipulating repressor <em>MKS1</em> is a promising approach to modulate yeast metabolism and to produce low-ethanol drinks.</p></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":null,"pages":null},"PeriodicalIF":4.1,"publicationDate":"2024-05-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S016816562400141X/pdfft?md5=1c6e8b47fd35c58a93531d4ba21567f8&pid=1-s2.0-S016816562400141X-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141071089","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"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

Preserved structure and function of human serum albumin self-folded in the oxidative cytoplasm of Escherichia coli 大肠杆菌氧化细胞质中自我折叠的人血清白蛋白的结构和功能保持不变

IF 4.1 2区生物学

Journal of biotechnology Pub Date : 2024-05-17 DOI: 10.1016/j.jbiotec.2024.05.005

Yong Joon Cho, Hyunji Kim, Sung In Lim

{"title":"Preserved structure and function of human serum albumin self-folded in the oxidative cytoplasm of Escherichia coli","authors":"Yong Joon Cho, Hyunji Kim, Sung In Lim","doi":"10.1016/j.jbiotec.2024.05.005","DOIUrl":"10.1016/j.jbiotec.2024.05.005","url":null,"abstract":"<div><p>Human serum albumin (HSA), a polypeptide featuring 17 disulfide bonds, acts as a crucial transport protein in human blood plasma. Its extended circulation half-life, mediated by FcRn (neonatal Fc receptor)-facilitated recycling, positions HSA as an excellent carrier for long-acting drug delivery. However, the conventional method of obtaining HSA from human blood faces limitations due to availability and potential contamination risks, such as blood-borne diseases. This study introduced SHuffle, an oxidative <em>Escherichia coli</em> (<em>E. coli</em>) expression system, for the production of recombinant HSA (rHSA) that spontaneously self-folds into its native conformation. This system ensures precise disulfide bond formation and correct folding of cysteine-rich rHSA, eliminating the need for chaperone co-expression or domain fusion of a folding enhancer. The purified rHSA underwent thorough physicochemical characterization, including mass spectrometry, circular dichroism spectroscopy, intrinsic fluorescence spectroscopy, esterase-like activity assay, and size exclusion chromatography, to assess critical quality attributes. Importantly, rHSA maintained native binding affinity to FcRn and the albumin-binding domain. Collectively, our analyses demonstrated a high comparability between rHSA and plasma-derived HSA. The expression of rHSA in <em>E. coli</em> with an oxidizing cytosol provides a secure and cost-effective approach, enhancing the potential of rHSA for diverse medical applications.</p></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":null,"pages":null},"PeriodicalIF":4.1,"publicationDate":"2024-05-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140957572","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"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

Applications of biocatalytic CC bond reductions in the synthesis of flavours and fragrances 生物催化 C=C 键还原在香精香料合成中的应用。

IF 4.1 2区生物学

Journal of biotechnology Pub Date : 2024-05-16 DOI: 10.1016/j.jbiotec.2024.05.006

Maria C. Cancellieri, Celeste Nobbio, Francesco G. Gatti, Elisabetta Brenna, Fabio Parmeggiani

{"title":"Applications of biocatalytic CC bond reductions in the synthesis of flavours and fragrances","authors":"Maria C. Cancellieri, Celeste Nobbio, Francesco G. Gatti, Elisabetta Brenna, Fabio Parmeggiani","doi":"10.1016/j.jbiotec.2024.05.006","DOIUrl":"10.1016/j.jbiotec.2024.05.006","url":null,"abstract":"<div><p>Industrial biotechnology and biocatalysis can provide very effective synthetic tools to increase the sustainability of the production of fine chemicals, especially flavour and fragrance (F&F) ingredients, the market demand of which has been constantly increasing in the last years. One of the most important transformations in F&F chemistry is the reduction of C<img>C bonds, typically carried out with metal-catalysed hydrogenations or hydride-based reagents. Its biocatalytic counterpart is a competitive alternative, showcasing a range of advantages such as excellent chemo-, regio- and stereoselectivity, ease of implementation, mild reaction conditions and modest environmental impact. In the present review, the application of biocatalysed alkene reductions (from microbial fermentations with wild-type strains to engineered isolated ene-reductase enzymes) to synthetic processes useful for the F&F industry will be described, highlighting not only the exquisite stereoselectivity achieved, but also the overall improvement when chirality is not involved. Multi-enzymatic cascades involving C<img>C bioreductions are also examined, which allow much greater chemical complexity to be built in one-pot biocatalytic systems.</p></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":null,"pages":null},"PeriodicalIF":4.1,"publicationDate":"2024-05-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S0168165624001342/pdfft?md5=9a15ae0b2ad22b505ec4aa4c319dbb70&pid=1-s2.0-S0168165624001342-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140957569","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"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

Optimizing interleukin-6 and 8 expression, clarification and purification in plant cell packs and plants for application in advanced therapy medicinal products and cellular agriculture 优化植物细胞包和植物中白细胞介素-6 和白细胞介素-8 的表达、澄清和纯化，以应用于高级治疗药物产品和细胞农业。

IF 4.1 2区生物学

Journal of biotechnology Pub Date : 2024-05-11 DOI: 10.1016/j.jbiotec.2024.05.003

P. Opdensteinen , J.F. Buyel

{"title":"Optimizing interleukin-6 and 8 expression, clarification and purification in plant cell packs and plants for application in advanced therapy medicinal products and cellular agriculture","authors":"P. Opdensteinen , J.F. Buyel","doi":"10.1016/j.jbiotec.2024.05.003","DOIUrl":"10.1016/j.jbiotec.2024.05.003","url":null,"abstract":"<div><p>Healthcare and nutrition are facing a paradigm shift in light of advanced therapy medicinal products (ATMPs) and cellular agriculture options respectively. Both options heavily rely on some sort of animal cell culture, e.g. autologous stem cells. These cultures require various growth factors, such as interleukin-6 and 8 (IL-6/8), in a pure, safe and sustainable form that can be provided in a scalable manner. Plants seem well suited for this task because purification of small proteins can be readily achieved by membrane separation, human/animal pathogens do not replicate in plants and production can be scaled up using in-door farming or agricultural practices. Here, we illustrate this capacity by first optimizing the codon usage of IL-6/8 for translation in <em>Nicotiana</em> spp., as well as testing the effect of untranslated regions and product targeting to different sub-cellular compartments on expression in a high-throughput plant cell pack (PCP) assay. In the chloroplast, IL-6 accumulated up to 6.9±3.8 (SD, n=2) and 14.4±7.4 mg kg<sup>−1</sup> (SD, n=5) were observed in case of IL-8. When transferring IL-8 expression into whole plants, accumulation was 12.3±1.5 mg kg<sup>−1</sup> (SD, n=3). After extraction and clarification, IL-8 was purified using a two-stage process consisting of an ultrafiltration/diafiltration step with 100 kDa and 10 kDa cut off membranes followed by an IMAC polishing step. The purity, yield and recovery were 97.8%, 6.6 mg kg<sup>−1</sup> and 38%, respectively. We evaluated the ability of the proposed purification process to remove endotoxins to ensure the compatibility of plant-made growth factors with cell culture.</p></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":null,"pages":null},"PeriodicalIF":4.1,"publicationDate":"2024-05-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S0168165624001317/pdfft?md5=8476c89d79ca8028884447f91574d459&pid=1-s2.0-S0168165624001317-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140916784","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"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

Exploring G-quadruplex structure in PRCC-TFE3 fusion oncogene: Plausible use as anti cancer therapy for translocation Renal cell carcinoma (tRCC) 探索PRCC-TFE3融合癌基因中的G-四叠体结构：可用作易位肾细胞癌（tRCC）的抗癌疗法。

IF 4.1 2区生物学

Journal of biotechnology Pub Date : 2024-05-11 DOI: 10.1016/j.jbiotec.2024.05.004

Neha Neha, Parimal Das

{"title":"Exploring G-quadruplex structure in PRCC-TFE3 fusion oncogene: Plausible use as anti cancer therapy for translocation Renal cell carcinoma (tRCC)","authors":"Neha Neha, Parimal Das","doi":"10.1016/j.jbiotec.2024.05.004","DOIUrl":"10.1016/j.jbiotec.2024.05.004","url":null,"abstract":"<div><p>The <em>TFE3</em> fusion gene, byproduct of Xp11.2 translocation, is the diagnostic marker for translocation renal cell carcinoma (tRCC). Absence of any clinically recognized therapy for tRCC, pressing a need to create novel and efficient therapeutic approaches. Previous studies shown that stabilization of the G-quadruplex structure in oncogenes suppresses their expression machinery. To combat the oncogenesis caused by fusion genes, our objective is to locate and stabilize the G-quadruplex structure within the <em>PRCC-TFE3</em> fusion gene. Using the Quadruplex-forming G Rich Sequences (QGRS) mapper and the Non-B DNA motif search tool (nBMST) online server, we found putative G-quadruplex forming sequences (PQS) in the <em>PRCC-TFE3</em> fusion gene. Circular dichroism demonstrating a parallel G-quadruplex in the targeted sequence. Fluorescence and UV–vis spectroscopy results suggest that pyridostatin binds to this newly discovered G-quadruplex. The PCR stop assay, as well as transcriptional or translational inhibition using real time PCR and Dual luciferase assay, revealed that stable G-quadruplex formation affects biological processes. Confocal microscopy of HEK293T cells transfected with the fusion transcript confirmed G-quadruplexes formation in cell. This investigation may shed light on G-quadruplex’s functions in fusion genes and may help in the development of therapies specifically targeted against fusion oncogenes, which would enhance the capability of current tRCC therapy approach.</p></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":null,"pages":null},"PeriodicalIF":4.1,"publicationDate":"2024-05-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140916783","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

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