Journal of biotechnology最新文献

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Exploration of new ways for CRISPR/Cas12a activation: DNA hairpins without PAM and toehold and single strands containing DNA and RNA bases 探索 CRISPR/Cas12a 激活的新途径:不含 PAM 和趾扣的 DNA 发夹以及含有 DNA 和 RNA 碱基的单链。
IF 4.1 2区 生物学
Journal of biotechnology Pub Date : 2024-06-14 DOI: 10.1016/j.jbiotec.2024.06.011
Wen He , Xinyu Li , Xinmin Li , Minghui Guo , Mengxuan Zhang , Ruiwei Hu , Menghan Li , Shijia Ding , Yurong Yan
{"title":"Exploration of new ways for CRISPR/Cas12a activation: DNA hairpins without PAM and toehold and single strands containing DNA and RNA bases","authors":"Wen He ,&nbsp;Xinyu Li ,&nbsp;Xinmin Li ,&nbsp;Minghui Guo ,&nbsp;Mengxuan Zhang ,&nbsp;Ruiwei Hu ,&nbsp;Menghan Li ,&nbsp;Shijia Ding ,&nbsp;Yurong Yan","doi":"10.1016/j.jbiotec.2024.06.011","DOIUrl":"10.1016/j.jbiotec.2024.06.011","url":null,"abstract":"<div><p>The CRISPR/Cas12a system is emerging as a promising candidate for next-generation diagnostic biosensing platforms, with the discovery of new activation modes greatly expanding its applications. Here, we have identified two novel CRISPR/Cas12a system activation modes: PAM- and toehold-free DNA hairpins, and DNA-RNA hybrid strands. Utilizing a well-established real-time fluorescence method, we have demonstrated a strong correlation between DNA hairpin structures and Cas12a activation. Compared with previously reported activation modes involving single-stranded DNA and PAM-contained double-stranded DNA, the DNA hairpin activation way exhibits similar specificity and generality. Moreover, our findings indicate that increasing the number of RNA bases in DNA-RNA hybrid strands can decelerate the kinetics of Cas12a-triggered <em>trans</em>-cleavage of reporter probes. These newly discovered CRISPR/Cas12a activation ways hold significant potential for the development of high-performance biosensing strategies.</p></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":null,"pages":null},"PeriodicalIF":4.1,"publicationDate":"2024-06-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141330964","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
Distal mutations enhance efficiency of free and immobilized NOV1 dioxygenase for vanillin synthesis 远端突变可提高游离和固定 NOV1 二氧合酶合成香兰素的效率。
IF 4.1 2区 生物学
Journal of biotechnology Pub Date : 2024-06-14 DOI: 10.1016/j.jbiotec.2024.06.012
Mario De Simone , Lur Alonso-Cotchico , Maria Fátima Lucas , Vânia Brissos , Lígia O. Martins
{"title":"Distal mutations enhance efficiency of free and immobilized NOV1 dioxygenase for vanillin synthesis","authors":"Mario De Simone ,&nbsp;Lur Alonso-Cotchico ,&nbsp;Maria Fátima Lucas ,&nbsp;Vânia Brissos ,&nbsp;Lígia O. Martins","doi":"10.1016/j.jbiotec.2024.06.012","DOIUrl":"10.1016/j.jbiotec.2024.06.012","url":null,"abstract":"<div><p>Protein engineering is crucial to improve enzymes’ efficiency and robustness for industrial biocatalysis. NOV1 is a bacterial dioxygenase that holds biotechnological potential by catalyzing the one-step oxidation of the lignin-derived isoeugenol into vanillin, a popular flavoring agent used in food, cleaning products, cosmetics and pharmaceuticals. This study aims to enhance NOV1 activity and operational stability through the identification of distal hotspots. located at more than 9 Å from the active site using Zymspot, a tool that predicts advantageous distant mutations, streamlining protein engineering. A total of 41 variants were constructed using site-directed mutagenesis and the six most active enzyme variants were then recombined. Two variants, with two and three mutations, showed nearly a 10-fold increase in activity and up to 40-fold higher operational stability than the wild-type. Furthermore, these variants show 90–100 % immobilization efficiency in metal affinity resins, compared to approximately 60 % for the wild-type. In bioconversions where 50 mM of isoeugenol was added stepwise over 24-h cycles, the 1D2 variant produced approximately 144 mM of vanillin after six reaction cycles, corresponding to around 22 mg, indicating a 35 % molar conversion yield. This output was around 2.5 times higher than that obtained using the wild-type. Our findings highlight the efficacy of distal protein engineering in enhancing enzyme functions like activity, stability, and metal binding selectivity, thereby fulfilling the criteria for industrial biocatalysts. This study provides a novel approach to enzyme optimization that could have significant implications for various biotechnological applications.</p></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":null,"pages":null},"PeriodicalIF":4.1,"publicationDate":"2024-06-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S0168165624001706/pdfft?md5=f1fc03f16d758621e215d4f3c4020cb6&pid=1-s2.0-S0168165624001706-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141330963","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
Tuning almond lipase features by the buffer used during immobilization: The apparent biocatalysts stability depends on the immobilization and inactivation buffers and the substrate utilized 通过固定过程中使用的缓冲液调整杏仁脂肪酶的特性:表面生物催化剂的稳定性取决于固定和失活缓冲液以及所使用的底物。
IF 4.1 2区 生物学
Journal of biotechnology Pub Date : 2024-06-13 DOI: 10.1016/j.jbiotec.2024.06.009
Oumaima Cherni , Diego Carballares , El Hocine Siar , Pedro Abellanas-Perez , Diandra de Andrades , Maria de Lourdes Teixeira de Moraes Polizeli , Javier Rocha-Martin , Sellema Bahri , Roberto Fernandez-Lafuente
{"title":"Tuning almond lipase features by the buffer used during immobilization: The apparent biocatalysts stability depends on the immobilization and inactivation buffers and the substrate utilized","authors":"Oumaima Cherni ,&nbsp;Diego Carballares ,&nbsp;El Hocine Siar ,&nbsp;Pedro Abellanas-Perez ,&nbsp;Diandra de Andrades ,&nbsp;Maria de Lourdes Teixeira de Moraes Polizeli ,&nbsp;Javier Rocha-Martin ,&nbsp;Sellema Bahri ,&nbsp;Roberto Fernandez-Lafuente","doi":"10.1016/j.jbiotec.2024.06.009","DOIUrl":"10.1016/j.jbiotec.2024.06.009","url":null,"abstract":"<div><p>The lipase from <em>Prunus dulcis</em> almonds was inactivated under different conditions. At pH 5 and 9, enzyme stability remained similar under the different studied buffers. However, when the inactivation was performed at pH 7, there were some clear differences on enzyme stability depending on the buffer used. The enzyme was more stable in Gly than when Tris was employed for inactivation. Then, the enzyme was immobilized on methacrylate beads coated with octadecyl groups at pH 7 in the presence of Gly, Tris, phosphate and HEPES. Its activity was assayed versus triacetin and <em>S</em>-methyl mandelate. The biocatalyst prepared in phosphate was more active versus <em>S</em>-methyl mandelate, while the other ones were more active versus triacetin. The immobilized enzyme stability at pH 7 depends on the buffer used for enzyme immobilization. The buffer used in the inactivation and the substrate used determined the activity. For example, glycine was the buffer that promoted the lowest or the highest stabilities depending on the substrate used to quantify the activities.</p></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":null,"pages":null},"PeriodicalIF":4.1,"publicationDate":"2024-06-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S0168165624001688/pdfft?md5=e36c97bc5bdf76b4de6b9dadab9c60e1&pid=1-s2.0-S0168165624001688-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141320867","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
Lipase-mediated alcoholysis for in situ production of ester bioaromas in licuri oil for cosmetic applications 以脂肪酶为介导的醇解技术,用于在地衣油中原位生产酯类生物芳香物质,并将其应用于化妆品中。
IF 4.1 2区 生物学
Journal of biotechnology Pub Date : 2024-06-12 DOI: 10.1016/j.jbiotec.2024.06.010
Rafael Chelala Moreira , Gislaine Ricci Leonardi , Juliano Lemos Bicas
{"title":"Lipase-mediated alcoholysis for in situ production of ester bioaromas in licuri oil for cosmetic applications","authors":"Rafael Chelala Moreira ,&nbsp;Gislaine Ricci Leonardi ,&nbsp;Juliano Lemos Bicas","doi":"10.1016/j.jbiotec.2024.06.010","DOIUrl":"10.1016/j.jbiotec.2024.06.010","url":null,"abstract":"<div><p>Bioaromas can be produced by lipases either through their hydrolytic or (trans)esterifying activities. Therefore, this work reports the development of a lipase-catalyzed biotransformed licuri oil, forming volatile ethyl esters with odor notes resembling tropical fruits. Ethyl octanoate formation was promoted when 7.0 % (m/v) Lipozyme 435® was used to convert a grain alcohol:licuri oil mixture (51:49, v/v) at 58ºC and 70 rpm for 6 hours. The biotransformed oil has shown antimicrobial activity against <em>Staphylococcus hominis</em>, <em>S. epidermidis,</em> and <em>Corynebacterium xerosis</em>, bacteria associated with bad skin odor. Finally, this biotransformed oil was used without further treatments (e.g., recovery or purification procedures) to prepare two cosmetic formulations (in a dosage of 1.5 %), aiming for both fragrant and deodorant activity.</p></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":null,"pages":null},"PeriodicalIF":4.1,"publicationDate":"2024-06-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141320866","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
Application of hydrophobic eutectic solvent in efficient biotransformation of total flavonoids of Herba Epimedii 疏水共晶溶剂在淫羊藿总黄酮高效生物转化中的应用
IF 4.1 2区 生物学
Journal of biotechnology Pub Date : 2024-06-11 DOI: 10.1016/j.jbiotec.2024.06.007
Qi Li , Shan Lu , Xianyao Wu , Lei Wang , Zhenzhong Wang , Linguo Zhao
{"title":"Application of hydrophobic eutectic solvent in efficient biotransformation of total flavonoids of Herba Epimedii","authors":"Qi Li ,&nbsp;Shan Lu ,&nbsp;Xianyao Wu ,&nbsp;Lei Wang ,&nbsp;Zhenzhong Wang ,&nbsp;Linguo Zhao","doi":"10.1016/j.jbiotec.2024.06.007","DOIUrl":"10.1016/j.jbiotec.2024.06.007","url":null,"abstract":"<div><p>Icaritin, a hydrolysate from total flavonoids of <em>Epimedii</em> (TFE), which has better anti-hepatoma activity than its glycosylated form. In this work, immobilized enzymes 4LP-Tpebgl3@Na-Y and DtRha@ES-107 were used to hydrolyze TFE to prepare icaritin. Five different hydrophobic deep eutectic solvents (HDES) were prepared and the most ideal HDES was successfully selected, which was composed of dodecyl alcohol and thymol with the molar ratio of 2:1. The relative enzyme activity of 4LP-Tpebgl3@Na-Y and DtRha@ES-107 was about 102.4 % and 112.5 %, respectively. In addition, the thermal and binding stability of 4LP-Tpebgl3@Na-Y and DtRha@ES-107 in HDES was not affected negatively. In the biphasic system composed of 50 % (<em>v</em>/<em>v</em>) HDES and Na<sub>2</sub>HPO<sub>4</sub>-citric acid buffer (50 mM, pH 5.5), 4LP-Tpebgl3@Na-Y (1.0 U/mL) and TFE (1 g/L) were reacted at 80 °C for 1 h, and then reacted with DtRha@ES-107 (20 U/mL) at 80 °C for 2 h. Finally, TFE was completely converted to 301.8 mg/L icaritin (0.82 mM). After 10 cycles, 4LP-Tpebgl3@Na-Y/DtRha@ES-107 still maintained 84.1 % original activity. In this study, we developed an efficient methodology for icaritin preparation through the integration of enzymatic catalysis and adsorption separation, presenting a viable approach for large-scale, cost-effective production of icaritin.</p></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":null,"pages":null},"PeriodicalIF":4.1,"publicationDate":"2024-06-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141317446","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
Molecular biomarkers identification and applications in CHO bioprocessing 分子生物标志物的鉴定和在 CHO 生物加工中的应用。
IF 4.1 2区 生物学
Journal of biotechnology Pub Date : 2024-06-08 DOI: 10.1016/j.jbiotec.2024.06.005
Caroline Desmurget , Arnaud Perilleux , Jonathan Souquet , Nicole Borth , Julien Douet
{"title":"Molecular biomarkers identification and applications in CHO bioprocessing","authors":"Caroline Desmurget ,&nbsp;Arnaud Perilleux ,&nbsp;Jonathan Souquet ,&nbsp;Nicole Borth ,&nbsp;Julien Douet","doi":"10.1016/j.jbiotec.2024.06.005","DOIUrl":"10.1016/j.jbiotec.2024.06.005","url":null,"abstract":"<div><p>Biomarkers are valuable tools in clinical research where they allow to predict susceptibility to diseases, or response to specific treatments. Likewise, biomarkers can be extremely useful in the biomanufacturing of therapeutic proteins. Indeed, constraints such as short timelines and the need to find hyper-productive cells could benefit from a data-driven approach during cell line and process development. Many companies still rely on large screening capacities to develop productive cell lines, but as they reach a limit of production, there is a need to go from empirical to rationale procedures. Similarly, during bioprocessing runs, substrate consumption and metabolism wastes are commonly monitored. None of them possess the ability to predict the culture behavior in the bioreactor. Big data driven approaches are being adapted to the study of industrial mammalian cell lines, enabled by the publication of Chinese hamster and CHO genome assemblies which allowed the use of next-generation sequencing with these cells, as well as continuous proteome and metabolome annotation. However, if these different -omics technologies contributed to the characterization of CHO cells, there is a significant effort remaining to apply this knowledge to biomanufacturing methods. The correlation of a complex phenotype such as high productivity or rapid growth to the presence or expression level of a specific biomarker could save time and effort in the screening of manufacturing cell lines or culture conditions. In this review we will first discuss the different biological molecules that can be identified and quantified in cells, their detection techniques, and associated challenges. We will then review how these markers are used during the different steps of cell line and bioprocess development, and the inherent limitations of this strategy.</p></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":null,"pages":null},"PeriodicalIF":4.1,"publicationDate":"2024-06-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S0168165624001640/pdfft?md5=e543988c7142b1b021941335f7d07fb1&pid=1-s2.0-S0168165624001640-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141296113","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
Kinetic and stability studies of amino acid metal-organic frameworks for encapsulating of amino acid dehydrogenase 用于封装氨基酸脱氢酶的氨基酸金属有机框架的动力学和稳定性研究
IF 4.1 2区 生物学
Journal of biotechnology Pub Date : 2024-06-07 DOI: 10.1016/j.jbiotec.2024.06.006
Lingling Dong , Yu Xiong , Xiaoyan Xiang , Feixuan Li , Qidi Song , Shizhen Wang
{"title":"Kinetic and stability studies of amino acid metal-organic frameworks for encapsulating of amino acid dehydrogenase","authors":"Lingling Dong ,&nbsp;Yu Xiong ,&nbsp;Xiaoyan Xiang ,&nbsp;Feixuan Li ,&nbsp;Qidi Song ,&nbsp;Shizhen Wang","doi":"10.1016/j.jbiotec.2024.06.006","DOIUrl":"https://doi.org/10.1016/j.jbiotec.2024.06.006","url":null,"abstract":"<div><p>Zr-MOFs was applied for the immobilization of hyperthermophilic and halophilic amino acid dehydrogenase (Zr-MOFs-NTAaDH) by physical adsorption for the biosynthesis of L-homophenylalanine. Activity of Zr-MOFs-NTAaDH was enhanced by 3.3-fold of the free enzyme at 70°C. And the enzyme activity of Zr-MOFs-NTAaDH was maintained at 4.16 U/mg at pH 11, which was 7.8 folds of that of NTAaDH. Kinetic parameters indicated catalytic efficiency of Zr-MOFs-NTAaDH was increased compared to the free enzyme as <em>k</em><sub><em>cat</em></sub> of Zr-MOFs-NTAaDH was 12.3-fold of that of free enzyme. After 7 recycles, the activity of Zr-MOFs-NTAaDH remained 68 %. And Zr-MOFs-NTAaDH exhibited high ionic liquid tolerance which indicated the great potential for industrial application.</p></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":null,"pages":null},"PeriodicalIF":4.1,"publicationDate":"2024-06-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141292057","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
Virion-surface display of a chimeric immunoglobulin Fc domain facilitating uptake by antigen-presenting cells 病毒表面显示的嵌合免疫球蛋白 Fc 结构域有助于抗原递呈细胞的吸收。
IF 4.1 2区 生物学
Journal of biotechnology Pub Date : 2024-06-06 DOI: 10.1016/j.jbiotec.2024.06.004
Sayuri Seki , Prince Kofi Parbie , Hiroyuki Yamamoto , Tetsuro Matano
{"title":"Virion-surface display of a chimeric immunoglobulin Fc domain facilitating uptake by antigen-presenting cells","authors":"Sayuri Seki ,&nbsp;Prince Kofi Parbie ,&nbsp;Hiroyuki Yamamoto ,&nbsp;Tetsuro Matano","doi":"10.1016/j.jbiotec.2024.06.004","DOIUrl":"10.1016/j.jbiotec.2024.06.004","url":null,"abstract":"<div><p>Antigen-presenting cells (APCs) play an important role in virus infection control by bridging innate and adaptive immune responses. Macrophages and dendritic cells (DCs) possess various surface receptors to recognize/internalize antigens, and antibody binding can enhance pathogen-opsonizing uptake by these APCs via interaction of antibody fragment crystallizable (Fc) domains with Fc receptors, evoking profound pathogen control in certain settings. Here, we examined phagocytosis-enhancing potential of Fc domains directly oriented on a retroviral virion/virus-like particle (VLP) surface. We generated an expression vector coding a murine Fc fragment fused to the transmembrane region (TM) of a retroviral envelope protein, deriving expression of the Fc-TM fusion protein on the transfected cell surface and production of virions incorporating the chimeric Fc upon co-transfection. Incubation of Fc-displaying simian immunodeficiency virus (SIV) with murine J774 macrophages and bone marrow-derived DCs derived Fc receptor-dependent enhanced uptake, being visualized by imaging cytometry. Alternative preparation of a murine leukemia virus (MLV) backbone-based Fc-displaying VLP loading an influenza virus hemagglutinin (HA) antigen resulted in enhanced HA internalization by macrophages, stating antigen compatibility of the design. Results show that the Fc-TM fusion molecule can be displayed on certain viruses/VLPs and may be utilized as a molecular adjuvant to facilitate APC antigen uptake.</p></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":null,"pages":null},"PeriodicalIF":4.1,"publicationDate":"2024-06-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141293445","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
Efficient production of a highly active lysozyme from European flat oyster Ostrea edulis 从欧洲平牡蛎(Ostrea edulis)中高效生产高活性溶菌酶。
IF 4.1 2区 生物学
Journal of biotechnology Pub Date : 2024-06-05 DOI: 10.1016/j.jbiotec.2024.05.011
Bo Pang , Manxi Song , Jiahao Yang , Haobin Mo , Kai Wang , Xia Chen , Yujun Huang , Ruixia Gu , Chengran Guan
{"title":"Efficient production of a highly active lysozyme from European flat oyster Ostrea edulis","authors":"Bo Pang ,&nbsp;Manxi Song ,&nbsp;Jiahao Yang ,&nbsp;Haobin Mo ,&nbsp;Kai Wang ,&nbsp;Xia Chen ,&nbsp;Yujun Huang ,&nbsp;Ruixia Gu ,&nbsp;Chengran Guan","doi":"10.1016/j.jbiotec.2024.05.011","DOIUrl":"10.1016/j.jbiotec.2024.05.011","url":null,"abstract":"<div><p>Lysozyme, an antimicrobial agent, is extensively employed in the food and healthcare sectors to facilitate the breakdown of peptidoglycan. However, the methods to improve its catalytic activity and secretory expression still need to be studied. In the present study, twelve lysozymes from different origins were heterologously expressed using the <em>Komagataella phaffii</em> expression system. Among them, the lysozyme from the European flat oyster <em>Ostrea edulis</em> (oeLYZ) showed the highest activity. Via a semi-rational approach to reduce the structural free energy, the double mutant Y15A/S39R (oeLYZ<sub>dm</sub>) with the catalytic activity 1.8-fold greater than that of the wild type was generated. Subsequently, different N-terminal fusion tags were employed to enhance oeLYZ<sub>dm</sub> expression. The fusion with peptide tag 6×Glu resulted in a remarkable increase in the recombinant oeLYZdm expression, from 2.81 × 10<sup>3</sup> U mL<sup>–1</sup> to 2.11 × 10<sup>4</sup> U mL<sup>–1</sup> in shake flask culture, and eventually reaching 2.05 × 10<sup>5</sup> U mL<sup>–1</sup> in a 3-L fermenter. The work produced the greatest amount of heterologous oeLYZ expression in microbial systems that are known to exist. Reducing the structural free energy and employing the N-terminal fusion tags are effective strategies to improve the catalytic activity and secretory expression of lysozyme.</p></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":null,"pages":null},"PeriodicalIF":4.1,"publicationDate":"2024-06-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141288158","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
Extension of the circulatory half-life of recombinant ecallantide via albumin fusion without loss of anti-kallikrein activity 通过白蛋白融合延长重组ecallantide的循环半衰期,同时不丧失抗卡力蛋白活性。
IF 4.1 2区 生物学
Journal of biotechnology Pub Date : 2024-06-04 DOI: 10.1016/j.jbiotec.2024.06.002
Ghofran Al-Adimi, Varsha Bhakta , Louise J. Eltringham-Smith , Valerie Shirobokov, William P. Sheffield
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