Journal of biotechnology最新文献_第8页

Engineering silica nanocoated whole-cell asymmetric biocatalyst for efficient preparation of a key chiral intermediate of (S)-Rivastigmine 工程二氧化硅纳米包被全细胞不对称生物催化剂高效制备(S)-利瓦斯汀关键手性中间体。

IF 4.1 2区生物学

Journal of biotechnology Pub Date : 2025-01-13 DOI: 10.1016/j.jbiotec.2025.01.005

Baoling Chen, Hang Yang, Ruixuan Bai, Xiaotong Du, Yue Gao, Liangyu Zheng

{"title":"Engineering silica nanocoated whole-cell asymmetric biocatalyst for efficient preparation of a key chiral intermediate of (S)-Rivastigmine","authors":"Baoling Chen, Hang Yang, Ruixuan Bai, Xiaotong Du, Yue Gao, Liangyu Zheng","doi":"10.1016/j.jbiotec.2025.01.005","DOIUrl":"10.1016/j.jbiotec.2025.01.005","url":null,"abstract":"<div><div>In our previous study, the whole cells containing an aldo–keto reductase (yhdN) and glucose dehydrogenase (GDH) were constructed and applied in a stereoselective carbonyl reduction reaction to prepare (<em>S</em>)-NEMCA-HEPE, being a key chiral intermediate of (<em>S</em>)-Rivastigmine which is widely prescribed for the treatment of Alzheimer’s disease. Although the conversion and enantiomeric excess (<em>e.e.</em>) could reach to 78.2 % and 99 %, respectively, ionic liquid as an additive was required to improve the permeability of cell membrane. To further simplify the reaction, the molecular docking and saturation mutagenesis technology were used here to obtain an activity-improved yhdN variant such as G19A. And then, both excellent conversion and <em>e.e.</em> of 99 % for (<em>S</em>)-NEMCA-HEPE could be achieved within 40 min by using only G19A-GDH whole cell as a catalyst without any additive. However, the use of the whole cells still faces the issues of poor operation stability and adverse application prospect. Subsequently, a hydrophobic \"cell-in-shell\" complex of G19A-GDH@O-Silica was constructed by using a silica nanocoated technology. The obtained G19A-GDH@O-Silica exhibited an excellent conversion towards the asymmetric carbonyl reduction, and a good tolerance in changing thermal, pH, and storage environmental. Giving 76.3 % of reaction conversion even after the 11th cycle of reuse, indicated that G19A-GDH@O-Silica also possessed ideal recyclability. The aim of this study is to provide a rapid, and cost-effective nanocoated whole-cell biocatalyst for efficient preparation of (<em>S</em>)-NEMCA-HEPE. The simplicity and robustness of the immobilization approach may become a powerful tool to utilize whole-cell catalysts towards organic catalysis.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"399 ","pages":"Pages 19-27"},"PeriodicalIF":4.1,"publicationDate":"2025-01-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143005990","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

Immobilization of glycosyltransferase into a hydrophilic metal-organic framework for efficient biosynthesis of chondroitin sulfate 糖基转移酶在亲水金属-有机框架中的固定化用于硫酸软骨素的高效生物合成。

IF 4.1 2区生物学

Journal of biotechnology Pub Date : 2025-01-10 DOI: 10.1016/j.jbiotec.2025.01.003

Xinyue Zhang , Yanqi Li , Jingjing Bi , Junjie Zhang , Bingzhi Li , Xing Zhang , Jie Zheng , Lei Lin

{"title":"Immobilization of glycosyltransferase into a hydrophilic metal-organic framework for efficient biosynthesis of chondroitin sulfate","authors":"Xinyue Zhang , Yanqi Li , Jingjing Bi , Junjie Zhang , Bingzhi Li , Xing Zhang , Jie Zheng , Lei Lin","doi":"10.1016/j.jbiotec.2025.01.003","DOIUrl":"10.1016/j.jbiotec.2025.01.003","url":null,"abstract":"<div><div>Chondroitin sulfate (CS) is a structurally complex anionic polysaccharide widely used in medical, cosmetic and food applications. Enzymatic catalysis is an important strategy for synthesizing CS with uniform chain lengths and well-defined structures. However, the industrial application of glycosyltransferases is hindered by limitations such as low expression yields, poor stability, and challenges in reuse. We developed a mild and rapid one-step synthetic method for the efficient immobilization of chondroitin synthase (KfoC). The resulting KfoC@ZIF-90 composite exhibits high catalytic activity, thermal stability, and pH adaptability. Notably, KfoC@ZIF-90 exhibited 5-fold enhanced thermal stability at 40°C and retained 86 % relative activity at pH 10, while also maintaining 90 % activity in organic solvents, surpassing the performance of free KfoC. Molecular docking analysis revealed that the binding capability of encapsulated KfoC with substrate was stronger than that of free KfoC, thereby improving catalytic performance. Furthermore, KfoC@ZIF-90 can be easily separated from the reaction solution by centrifugation, simplifying product isolation and purification while enabling enzyme reuse. These attributes significantly enhance operability and reduce processing costs, making enzymatic CS synthesis more feasible for industrial applications.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"399 ","pages":"Pages 63-71"},"PeriodicalIF":4.1,"publicationDate":"2025-01-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142970698","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

Assessing the role of deep eutectic solvents in Yarrowia lipolytica inhibition 评估深共晶溶剂在抑制多脂耶氏菌中的作用

IF 4.1 2区生物学

Journal of biotechnology Pub Date : 2024-11-28 DOI: 10.1016/j.jbiotec.2024.11.016

Filipe S. Buarque , Bernardo D. Ribeiro , Mara G. Freire , Maria A.Z. Coelho , Matheus M. Pereira

{"title":"Assessing the role of deep eutectic solvents in Yarrowia lipolytica inhibition","authors":"Filipe S. Buarque , Bernardo D. Ribeiro , Mara G. Freire , Maria A.Z. Coelho , Matheus M. Pereira","doi":"10.1016/j.jbiotec.2024.11.016","DOIUrl":"10.1016/j.jbiotec.2024.11.016","url":null,"abstract":"<div><div><em>Yarrowia lipolytica</em> has gained recognition as a microorganism with biological relevance and extensive biotechnological applications. Some of its features include a high enzyme secretion capacity and a high cell-density fermentation mode. Hexokinase (YlHxk) is a vital enzyme in <em>Y. lipolytica</em> growth since it catalyzes glucose metabolism through phosphorylation in the glycolytic pathway. Given the potential application of deep eutectic solvents (DES) as novel solvents in biotechnological processes, this study evaluated the influence of eighteen DES on the growth of <em>Y. lipolytica</em>. Furthermore, this work examined the effects of individual ions on the YlHxk enzyme by analyzing its enzymatic tunnel structure, molecule transport, and molecular docking. The results revealed a significant reduction in yeast growth in the presence of most DES compared to the control (medium without DES), with the exception of the [N<sub>8881</sub>]Cl: hexanoic acid (1:1) DES. The growth varied between 11.95 ± 0.60 and 0.68 ± 0.17 g dry cell weight L<sup>−1</sup>. According to the enzymatic tunnel analysis, DES components associated with the lowest microbial growth values were transported through tunnel 1. On the other hand, DES components had their pathway facilitated through tunnel 2 ([N<sub>8881</sub>]<sup>+</sup> and hexanoic acid) and showed growth values close to the control. Molecular docking analysis identified a similarity between all the ligands in this tunnel (including substrate and product), presenting binding interactions with the ASN273 amino acid of the YlHxk active site. Combining experimental results with computational tools provided promising insights at the molecular level, while also potentially reducing analysis costs and time, paving the way for similar approaches in broad biocatalytic reactions.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"398 ","pages":"Pages 1-10"},"PeriodicalIF":4.1,"publicationDate":"2024-11-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142748017","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

Creation of catalytic activity-improved hyperthermophilic PQQ-dependent aldose sugar dehydrogenase and its efficient use for high performance electro-device 催化活性改进的超耐热pqq依赖性醛糖脱氢酶的建立及其在高性能电器件中的高效应用。

IF 4.1 2区生物学

Journal of biotechnology Pub Date : 2024-11-28 DOI: 10.1016/j.jbiotec.2024.11.017

Miku Maeno , Yusuke Miki , Kazuki Ito , Haruhiko Sakuraba , Toshihisa Ohshima , Shin-ichiro Suye , Takenori Satomura

{"title":"Creation of catalytic activity-improved hyperthermophilic PQQ-dependent aldose sugar dehydrogenase and its efficient use for high performance electro-device","authors":"Miku Maeno , Yusuke Miki , Kazuki Ito , Haruhiko Sakuraba , Toshihisa Ohshima , Shin-ichiro Suye , Takenori Satomura","doi":"10.1016/j.jbiotec.2024.11.017","DOIUrl":"10.1016/j.jbiotec.2024.11.017","url":null,"abstract":"<div><div>PQQ-dependent aldose sugar dehydrogenase (PQQ-ASD) from the hyperthermophilic archaeon <em>Pyrobaculum aerophilum</em> (PaeASD) has great potential as an element for durable bioelectrodevices owing to its exceptional stability against high temperatures and across a broad pH spectrum. However, its application is constrained by low electric current output of the enzyme-immobilized electrodes, which is attributable to its low catalytic activity. A directed evolutionary approach was performed on PaeASD to improve enzyme activity, resulting in the identification of a PaeASD s24 mutant containing six amino acid substitutions, which exhibited a 16-fold higher specific activity than that of wild type. Although each single amino acid mutant among these substitutions exhibited lower enzyme activity than PaeASD s24, the double mutant R64Q/D350N showed enzyme activity comparable to that of PaeASD s24. These amino acids located in the vicinity of coenzyme PQQ within the PaeASD molecule are also highly conserved with those of PQQ-ASDs reported to date. Thus, these amino acids play crucial roles in the catalytic activity of PQQ-ASD. Furthermore, the <em>K</em><sub>m</sub> value for <span>d</span>-glucose of PaeASD s24-immobilized electrode decreased to approximately 1/3 that of the wild-type-immobilized electrode. These results indicate that the PaeASD s24 mutant is an excellent catalyst for potential bioelectrodevice applications.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"398 ","pages":"Pages 11-17"},"PeriodicalIF":4.1,"publicationDate":"2024-11-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142755169","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

Mechanism of rhamnolipid promoting the degradation of polycyclic aromatic hydrocarbons by gram-positive bacteria—Enhance transmembrane transport and electron transfer 鼠李糖脂促进革兰氏阳性菌降解多环芳烃的机制--增强跨膜转运和电子传递

IF 4.1 2区生物学

Journal of biotechnology Pub Date : 2024-11-23 DOI: 10.1016/j.jbiotec.2024.11.010

Bo Zhang , Lei Wang , Zhenjun Diwu , Maiqian Nie , Hongyun Nie

{"title":"Mechanism of rhamnolipid promoting the degradation of polycyclic aromatic hydrocarbons by gram-positive bacteria—Enhance transmembrane transport and electron transfer","authors":"Bo Zhang , Lei Wang , Zhenjun Diwu , Maiqian Nie , Hongyun Nie","doi":"10.1016/j.jbiotec.2024.11.010","DOIUrl":"10.1016/j.jbiotec.2024.11.010","url":null,"abstract":"<div><div>In this study, the Gram-positive bacterium <em>Bacillus licheniformis</em> T5 was utilized to investigate the impact of rhamnolipid on cell membrane and cell wall, as well as enzyme activity and electron transfer rate within cells. Results indicated that at the optimal concentration of rhamnolipid (200 mg/L), the cell membrane protein and cell wall peptidoglycan content of T5 decreased significantly. Infrared spectrum analysis and ultrastructure observations confirmed these findings, revealing noticeable changes in cell morphology in the presence of rhamnolipid. Specifically, cell folds increased, cell wall texture loosened, thickness decreased sharply, transmembrane channels appeared, and the plasma wall slightly separated. These alterations likely contributed to the increased permeability of the cell membrane. Furthermore, rhamnolipid accelerated the electron transfer rate in T5 cells, enhancing oxidoreductase activity. This study elucidates the mechanism through which rhamnolipid promotes the degradation of polycyclic aromatic hydrocarbons by Gram-positive bacteria, focusing on transmembrane transport and catalytic metabolism.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"397 ","pages":"Pages 51-60"},"PeriodicalIF":4.1,"publicationDate":"2024-11-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142706753","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

Engineering Saccharomyces boulardii for cell surface display of heterologous protein 改造布拉氏酵母菌，使其在细胞表面显示异源蛋白。

IF 4.1 2区生物学

Journal of biotechnology Pub Date : 2024-11-20 DOI: 10.1016/j.jbiotec.2024.11.013

Jamin Shin , Gayoung Lee , Won-Jae Chi , Sujeong Park , Yong-Su Jin , Soo Rin Kim

{"title":"Engineering Saccharomyces boulardii for cell surface display of heterologous protein","authors":"Jamin Shin , Gayoung Lee , Won-Jae Chi , Sujeong Park , Yong-Su Jin , Soo Rin Kim","doi":"10.1016/j.jbiotec.2024.11.013","DOIUrl":"10.1016/j.jbiotec.2024.11.013","url":null,"abstract":"<div><div><em>Saccharomyces boulardii</em> and <em>Saccharomyces cerevisiae</em> share over 99 % genetic similarity yet exhibit distinct metabolic traits. While the cell surface display system of <em>S. cerevisiae</em> is well-documented, the equivalent system in <em>S. boulardii</em> has yet to be fully characterized. This study investigates the cell surface display system of <em>S. boulardii</em> for the expression of a heterologous protein using different anchor proteins. Six strains expressing the enhanced green fluorescent protein (Egfp) and an anchor protein as a fusion protein were constructed to visualize the cell surface display system. Then a heterologous endo-inulinase protein was expressed with selected anchor proteins through fluorescence intensity comparison. Analysis by fluorescence microscopy revealed that the anchor protein Sed1 exhibited the highest fluorescence intensity. Furthermore, expressed selected anchor proteins and heterologous protein, endo-inulinase, the engineered strain could degrade and consume almost inulin in 72 h. Through endo-inulinase expression, we confirmed that not only Egfp but also heterologous protein is well expressed, and we successfully built an <em>S. boulardii</em> cell surface display system.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"397 ","pages":"Pages 44-50"},"PeriodicalIF":4.1,"publicationDate":"2024-11-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142692964","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

Comparative analysis of substrate- and regio-selectivity of HpaB monooxygenases and their application to hydroxydaidzein synthesis HpaB 单加氧酶底物和区域选择性的比较分析及其在羟基蝙蝠葛素合成中的应用。

IF 4.1 2区生物学

Journal of biotechnology Pub Date : 2024-11-20 DOI: 10.1016/j.jbiotec.2024.11.012

Sachiko Watanabe , Hideki Kato , Kento Yoshinaga , Akiko Kohara , Yuichi Ukawa , Akinobu Matsuyama , Toshiki Furuya

{"title":"Comparative analysis of substrate- and regio-selectivity of HpaB monooxygenases and their application to hydroxydaidzein synthesis","authors":"Sachiko Watanabe , Hideki Kato , Kento Yoshinaga , Akiko Kohara , Yuichi Ukawa , Akinobu Matsuyama , Toshiki Furuya","doi":"10.1016/j.jbiotec.2024.11.012","DOIUrl":"10.1016/j.jbiotec.2024.11.012","url":null,"abstract":"<div><div>4-Hydroxyphenylacetate 3-hydroxylase (HpaB) has high potential for use in polyphenol synthesis via <em>ortho</em>-hydroxylation. Although the HpaB enzymes from <em>Pseudomonas aeruginosa</em> (PaHpaB) and <em>Escherichia coli</em> (EcHpaB) have been well studied, few studies have compared their activity and substrate selectivity. Thus, which HpaB is optimal for use in the biotechnological production of polyphenols is unclear. In this study, we performed a comparative analysis of the substrate- and regio-selectivity of PaHpaB, EcHpaB, and the recently discovered enzyme from <em>Rhodococcus opacus</em> (RoHpaB). The activity of these enzymes was first compared toward representative aromatic substrates. PaHpaB and EcHpaB exhibited very similar catalytic activity toward <em>p</em>-coumaric acid and tyrosol with one benzene ring, whereas PaHpaB exhibited greater activity than EcHpaB toward resveratrol and naringenin with two benzene rings. These results suggest that PaHpaB is superior to EcHpaB in converting bulky compounds. Furthermore, PaHpaB also exhibited catalytic activity toward a flavonoid, daidzein (7,4′-dihydroxyisoflavone), whereas EcHpaB did not. RoHpaB also exhibited strong activity toward daidzein in addition to other aromatic substrates. Interestingly, PaHpaB hydroxylated the 6-position of daidzein, whereas RoHpaB hydroxylated the 3′-position. PaHpaB and RoHpaB enabled the facile synthesis of not only 6-hydroxydaidzein and 3′-hydroxydaidzein but also 6,3′-dihydroxydaidzein via the cascade reaction. This study is the first to demonstrate synthesis of hydroxydaidzeins using HpaB enzymes.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"397 ","pages":"Pages 61-66"},"PeriodicalIF":4.1,"publicationDate":"2024-11-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142692963","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

Digital holographic microscopy is suitable for lipid accumulation analysis in single cells of Yarrowia lipolytica 数字全息显微镜适用于分析脂溶性亚罗菌单细胞中的脂质积累。

IF 4.1 2区生物学

Journal of biotechnology Pub Date : 2024-11-17 DOI: 10.1016/j.jbiotec.2024.11.011

Simon Carl-Philipp Briel , Nicolas Feuser , Eva Johanna Moldenhauer , Johannes Kabisch , Peter Neubauer , Stefan Junne

{"title":"Digital holographic microscopy is suitable for lipid accumulation analysis in single cells of Yarrowia lipolytica","authors":"Simon Carl-Philipp Briel , Nicolas Feuser , Eva Johanna Moldenhauer , Johannes Kabisch , Peter Neubauer , Stefan Junne","doi":"10.1016/j.jbiotec.2024.11.011","DOIUrl":"10.1016/j.jbiotec.2024.11.011","url":null,"abstract":"<div><div>Digital holographic microscopy (DHM) is a label-free analytical technique for the determination of the cells’ volume and their cytosolic refractive index. Here, we demonstrate the suitability of DHM for the quantification of total lipid accumulation in the oleaginous yeast <em>Yarrowia lipolytica</em>. Presently, microbial lipids are gaining increasing attention due to their nutritional value in feed and food applications. Their microbiological synthesis in algae and yeast is subject to optimization studies, which necessitates rapid quantification of total lipids for faster progress and the possibility of process control. So far, quantification of the total intracellular long-chain fatty acid concentration in yeast cells is time-consuming though when common chromatography for a volumetric analysis or staining and flow cytometry for a single-cell based analysis are used. This study, however, demonstrates that 3D-DHM facilitates a quasi-real-time measurement that allows for a rapid quantification of total intracellular lipid accumulation on a single-cell level without cell staining. Data from wild-type and lipid overproducing <em>Y. lipolytica</em> strains with specific yields of long-chain fatty acids in a range between 70 and 360 mg/gCDW show a good correlation with the optical volume determined by DHM, as the total lipid accumulation in the cell is typically well correlated with the long-chain fatty acid concentration. The results further correlate with data obtained from gas chromatography and flow cytometry of Nile Red-stained cells, which proves the reliability of DHM for lipid quantification in <em>Y. lipolytica</em>.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"397 ","pages":"Pages 32-43"},"PeriodicalIF":4.1,"publicationDate":"2024-11-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142648166","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

A pump-free microfluidic co-culture system for investigating NK cell-tumor spheroid interactions in flow conditions 用于研究流动条件下 NK 细胞与肿瘤球状体相互作用的无泵微流体共培养系统

IF 4.1 2区生物学

Journal of biotechnology Pub Date : 2024-11-15 DOI: 10.1016/j.jbiotec.2024.11.008

Yuanyuan Xie , Ke Ning , Wen Sun , Lingke Feng , Yirong Chen , Wei Sun , Yan Li , Ling Yu

{"title":"A pump-free microfluidic co-culture system for investigating NK cell-tumor spheroid interactions in flow conditions","authors":"Yuanyuan Xie , Ke Ning , Wen Sun , Lingke Feng , Yirong Chen , Wei Sun , Yan Li , Ling Yu","doi":"10.1016/j.jbiotec.2024.11.008","DOIUrl":"10.1016/j.jbiotec.2024.11.008","url":null,"abstract":"<div><div>Natural killer (NK) cells are pivotal in immunotherapy due to their potent tumor-targeting capabilities. However, accessible <em>in vitro</em> 3D dynamic models for evaluating Tumor Infiltrating Natural Killer Cells (TINKs) remain scarce. This study addresses this gap by developing a novel pump-free microfluidic chip to investigate the interactions between NK-92 cells and prostate DU 145 tumor spheroids. The platform facilitates the separation of free NKs and TINKs for subtype characterization. The design integrates multiple planes with a multi-layer paper scaffold to accommodate tumor spheroids, allowing NK-92 cells to traverse Matrigel-coated barriers that mimic the extracellular matrix. The dual-channel pump-free device enables unidirectional circulation of NK-92 cells, allowing analysis of tumor spheroid movement and NK-92 cell interactions under flow conditions. Results demonstrate continuous fluid circulation in the dual-channel device by rocking the platform at tilt angles of 21° and 15°. Tumor spheroids show- enhanced migration under flow conditions compared to static culture. Although spheroids recruit slightly more NK-92 cells under flow conditions, CD56 and CD16 receptor expression on IL-2-activated free NK-92 cells and tumor-infiltrating NK-92 cells matches <em>in vivo</em> patterns in dynamic cultures. These findings suggest that tumor cells and fluid dynamics significantly influence NK cell subtypes. This pump-free microfluidic platform is a functional tool for simulating and studying immune cell-tumor interactions, providing valuable insights into NK cell dynamics with tumor spheroids in physiologically relevant environments.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"397 ","pages":"Pages 11-21"},"PeriodicalIF":4.1,"publicationDate":"2024-11-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142644206","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

Engineering Saccharomyces cerevisiae for continuous secretory production of hEGF in biofilm 对酿酒酵母进行工程改造，使其能在生物膜中持续分泌生产 hEGF。

IF 4.1 2区生物学

Journal of biotechnology Pub Date : 2024-11-15 DOI: 10.1016/j.jbiotec.2024.11.007

Kaiqi Zhi , Xiang Zhou , Tianping Gao , Kehan Liu , Zhenyu Wang , Yafan Cai , Zhi Wang , Shilei Wang , Jinle Liu , Dong Liu , Hanjie Ying

{"title":"Engineering Saccharomyces cerevisiae for continuous secretory production of hEGF in biofilm","authors":"Kaiqi Zhi , Xiang Zhou , Tianping Gao , Kehan Liu , Zhenyu Wang , Yafan Cai , Zhi Wang , Shilei Wang , Jinle Liu , Dong Liu , Hanjie Ying","doi":"10.1016/j.jbiotec.2024.11.007","DOIUrl":"10.1016/j.jbiotec.2024.11.007","url":null,"abstract":"<div><div>Human epidermal growth factor (hEGF) plays a crucial role in promoting cell growth and has various clinical applications. Due to limited natural sources and the high cost of chemical synthesis, researchers are now exploring genetic engineering as a potential method for hEGF production. In this particular study, a novel hEGF expression system was developed using <em>Saccharomyces cerevisiae</em>. This system involved optimizing the promoter and signal peptide and deleting protease-coding genes <em>PEP4</em>, <em>PRB1</em>, and <em>YAP3</em>, overexpressing chaperones <em>KAR2</em> and <em>PDI1</em> in the protein secretion pathway, which led to a 2.01-fold increase in hEGF production compared to the wild type strain. Furthermore, biofilm-forming genes <em>FLO11</em> and <em>ALS3</em> were integrated to create a biofilm strain with adhesive properties. A biofilm-based immobilized continuous fermentation model was established to leverage the characteristics of this biofilm strain. Each batch of this model yielded 130 mg/L of hEGF, with a production efficiency of 2.71 mg/L/h - surpassing the production efficiency of traditional free fermentation (1.62 mg/L/h). This study presents a promising fermentation model for efficient hEGF production based on biofilm characteristics, offering valuable insights for the application of biofilm fermentation in the production of small molecule peptides.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"397 ","pages":"Pages 1-10"},"PeriodicalIF":4.1,"publicationDate":"2024-11-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142644207","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