Journal of biotechnology最新文献_第2页

Generation of VacA-targeting guide peptides to increase specific antimicrobial peptide toxicity against Helicobacter pylori

IF 4.1 2区生物学

Journal of biotechnology Pub Date : 2025-03-27 DOI: 10.1016/j.jbiotec.2025.03.018

Patrick S. Ortiz, Mikaeel Young, Toslim Mahmud, Md. Mehadi Hasan Sohag, Christopher M. Kearney

{"title":"Generation of VacA-targeting guide peptides to increase specific antimicrobial peptide toxicity against Helicobacter pylori","authors":"Patrick S. Ortiz, Mikaeel Young, Toslim Mahmud, Md. Mehadi Hasan Sohag, Christopher M. Kearney","doi":"10.1016/j.jbiotec.2025.03.018","DOIUrl":"10.1016/j.jbiotec.2025.03.018","url":null,"abstract":"<div><h3>Background</h3><div>The <em>H. pylori</em> virulence factors VacA and CagA are the primary determinants of gastric cancer globally. In this study we increased the activity of antimicrobial peptides (AMPs) against <em>H. pylori</em> by using phage display against VacA to rapidly generate peptides targeting VacA, subsequently fusing these peptides to the AMP N-terminus to confer specificity.</div></div><div><h3>Results</h3><div>After four rounds of phage display, 36 phage clones were ranked for whole cell <em>H. pylori</em> binding by ELISA. The displayed 12-mer peptides of the top nine candidate clones were fused to GFP as guide peptides and analyzed for binding to wild type <em>H. pylori</em> 60190 and a ∆<em>vacA</em> strain. The three guides with the best differential binding were fused to the AMP pexiganan using two different linker peptides. All guide/linker combinations led to increased toxicity against <em>H. pylori</em> and most also decreased off-target toxicity. Guide P5 linked to pexiganan was the top configuration, delivering 64- to > 256-fold differential toxicity against <em>H. pylori</em> compared to off-target bacteria and a therapeutic window exceeding 128-fold when tested against cultured gastric cells.</div></div><div><h3>Conclusions</h3><div>Guide peptide biopanning provides an effective, scalable method to increase specific activity of antimicrobial peptides based on attraction to a key virulence factor.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"403 ","pages":"Pages 17-29"},"PeriodicalIF":4.1,"publicationDate":"2025-03-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143738143","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

Enhancing nicotinamide mononucleotide production in Escherichia coli through systematic metabolic engineering

IF 4.1 2区生物学

Journal of biotechnology Pub Date : 2025-03-27 DOI: 10.1016/j.jbiotec.2025.03.014

Zhaoyuan Zhang , Jiehu Liu , Meng Wang , Yang Li , Minglei Hou , Jiaren Cao , Jing Wu , Lingqia Su

{"title":"Enhancing nicotinamide mononucleotide production in Escherichia coli through systematic metabolic engineering","authors":"Zhaoyuan Zhang , Jiehu Liu , Meng Wang , Yang Li , Minglei Hou , Jiaren Cao , Jing Wu , Lingqia Su","doi":"10.1016/j.jbiotec.2025.03.014","DOIUrl":"10.1016/j.jbiotec.2025.03.014","url":null,"abstract":"<div><div>Nicotinamide mononucleotide (NMN) serves as a crucial precursor in the biosynthesis of NAD<sup>+</sup> and has garnered significant attention in the food, dietary supplement, and cosmetic industries. This study engineered an <em>Escherichia coli</em> strain for enhancing NMN production. Firstly, the strain with reduced NMN degradation and the ability to transport NMN extracellularly was constructed. Meanwhile, the gene encoding nicotinamide phosphoribosyltransferase (<em>pncA</em>) was disrupted to minimize substrate nicotinamide (NAM) degradation. Then, the induction starting point was optimized to alleviate the metabolic burden on the engineered strain. Subsequently, systematic remodeling of <em>E. coli</em>'s glucose metabolism was conducted to enhance its suitability for NMN production by overexpressing key enzymes of the pentose phosphate pathway (Zwf and Gnd), knocking out genes related to the Entner-Doudoroff pathway (<em>gntR</em> and <em>edd</em>), and further attenuating the glycolytic pathway. Then, we concentrated on optimizing the cellular metabolic state, meticulously balancing intracellular redox homeostasis. Finally, using glucose and 2 g/L of NAM as substrates, the extracellular NMN yield reached 4.96 g/L, which is the highest yield reported so far in similar research. These findings contribute to the commercial production of NMN and offer valuable insights for constructing efficient cell factories for other nucleotide compounds.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"403 ","pages":"Pages 73-80"},"PeriodicalIF":4.1,"publicationDate":"2025-03-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143742131","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

Microbial production of hyaluronic acid: The current advances, engineering strategies and trends

IF 4.1 2区生物学

Journal of biotechnology Pub Date : 2025-03-26 DOI: 10.1016/j.jbiotec.2025.03.015

Ehab Marwan-Abdelbaset , Mohamed Samy-Kamal , Dan Tan , XiaoYun Lu

{"title":"Microbial production of hyaluronic acid: The current advances, engineering strategies and trends","authors":"Ehab Marwan-Abdelbaset , Mohamed Samy-Kamal , Dan Tan , XiaoYun Lu","doi":"10.1016/j.jbiotec.2025.03.015","DOIUrl":"10.1016/j.jbiotec.2025.03.015","url":null,"abstract":"<div><div>Hyaluronic acid (HA) is a versatile biomolecule with applications in medicine, cosmetics, and pharmaceuticals. While traditionally extracted from animal tissues, HA is now predominantly produced through microbial fermentation. Microbial fermentation using strains such as <em>Streptococcus zooepidemicus</em>, <em>Corynebacterium glutamicum</em>, and <em>Bacillus subtilis</em> offers a more scalable and sustainable alternative to chemical and animal extraction methods. Recent studies reveal promising yields from engineered strains of <em>Corynebacterium glutamicum</em> and <em>Bacillus subtilis</em>, utilizing advanced metabolic and genetic techniques. Recent advancements in genetic and metabolic engineering, as well as synthetic biology, have addressed some challenges related to molecular weight, viscosity, and by-product formation. This review focuses on the microbial production of HA using engineered strains, encompassing producer organisms, metabolic engineering strategies, industrial-scale production, and key factors influencing molecular weight. Furthermore, it addresses the challenges and potential solutions associated with HA production. Additional research is necessary to develop more efficient and robust engineered strains that exhibit resistance to contamination and can utilize low-cost substrates, such as <em>Pseudomonas putida</em> and <em>Halomonas</em> spp. By overcoming these challenges, researchers can advance the industrial production of HA and expand its applications, thereby contributing to the growth of the HA market.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"403 ","pages":"Pages 52-72"},"PeriodicalIF":4.1,"publicationDate":"2025-03-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143742930","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

Production and purification of recombinant long protein isoforms of FGF11 subfamily

IF 4.1 2区生物学

Journal of biotechnology Pub Date : 2025-03-26 DOI: 10.1016/j.jbiotec.2025.03.016

Martyna Biadun , Szymon Sidor , Marta Kalka , Radoslaw Karelus , Martyna Sochacka , Daniel Krowarsch , Lukasz Opalinski , Malgorzata Zakrzewska

引用次数: 0

Harnessing recombinant Bacillus licheniformis CotA laccase for electrochemical detection of catechol

IF 4.1 2区生物学

Journal of biotechnology Pub Date : 2025-03-25 DOI: 10.1016/j.jbiotec.2025.03.017

Stanzin Lzaod , Sumit Sharma , Samaresh Das , Tanmay Dutta

{"title":"Harnessing recombinant Bacillus licheniformis CotA laccase for electrochemical detection of catechol","authors":"Stanzin Lzaod , Sumit Sharma , Samaresh Das , Tanmay Dutta","doi":"10.1016/j.jbiotec.2025.03.017","DOIUrl":"10.1016/j.jbiotec.2025.03.017","url":null,"abstract":"<div><div>Laccases, known for their ability to oxidize a broad range of substrates and catalyze multiple reactions, offer tremendous potential for varied applications. Despite their widespread presence in nature, research has primarily focused on fungal laccases. However, fungal laccases are susceptible to extreme conditions and inhibitors, hindering their widespread industrial use. Under such circumstances, a burgeoning interest has surrounded extremophilic and cost-effective bacterial laccases. Consequently, we explored the potential of recombinant <em>Bacillus licheniformis</em> laccase (CotA) in the fabrication of an electrochemical biosensor for the detection of catechol, an environmental pollutant. The biosensor was constructed by modifying a screen-printed electrode with CotA encapsulated in a conducting polymer (PEDOT:PSS)/chitosan film. CotA can oxidize catechol, and this step enabled the detection of catechol through amperometric measurements. The biosensor demonstrated competitive analytical features to fungal laccases with a low detection limit (1.4μM), high sensitivity (42.637 μAmM<sup>−1</sup>) and excellent storage stability retaining 90 % of its initial activity after 40 days of storage at 4 °C. Furthermore, it successfully detected catechol in spiked tap and river water samples making it an effective and efficient solution for monitoring catechol in real environmental samples.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"403 ","pages":"Pages 30-39"},"PeriodicalIF":4.1,"publicationDate":"2025-03-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143730109","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

Retraction notice to “A recombinant H1 histone based system for efficient delivery of nucleic acids” [J. Biotechnol. 105 (2003) 215–226]

IF 4.1 2区生物学

Journal of biotechnology Pub Date : 2025-03-23 DOI: 10.1016/j.jbiotec.2025.03.003

Iratxe Puebla , Selma Esseghir , Alison Mortlock , Anthony Brown , Andrea Crisanti , Walter Low

引用次数: 0

Comparative characterisation of autotrophic and heterotrophic isopropanol formation by Cupriavidus necator in shake flasks

IF 4.1 2区生物学

Journal of biotechnology Pub Date : 2025-03-20 DOI: 10.1016/j.jbiotec.2025.03.011

I. Weickardt , E. Lombard , A. Zhang , L. Blank , S.E. Guillouet

{"title":"Comparative characterisation of autotrophic and heterotrophic isopropanol formation by Cupriavidus necator in shake flasks","authors":"I. Weickardt , E. Lombard , A. Zhang , L. Blank , S.E. Guillouet","doi":"10.1016/j.jbiotec.2025.03.011","DOIUrl":"10.1016/j.jbiotec.2025.03.011","url":null,"abstract":"<div><div>Autotrophic cultivation offers a path to carbon-neutral bioproduction, which is increasingly valuable in the context of climate change mitigation. In this study, the production of isopropanol by <em>Cupriavidus necator</em> is used as an example for CO<sub>2</sub> valorisation, and a simple shake bottle system is introduced to facilitate the development of aerobic autotrophic cultivation processes and strain screening. Applying 1.5 bar overpressure in the bottle's headspace enhances gas transfer while pressure decrease was shown to be correlated to biomass and product formation, allowing to follow metabolic activity without sampling. After optimizing cultivation parameters and nickel feeding strategy, the system was applied to compare three different isopropanol-producing strains. The highest autotrophically obtained isopropanol concentration was 2.2 ± 0.5 g L<sup>−1</sup> with a specific yield of 0.9 ± 0.2 g g<sub>CDW</sub><sup>−1</sup> and a minimal by-product concentration of 0.05 ± 0.01 g L<sup>−1</sup> acetone. Heterotrophic cultivations were carried out for comparison, obtaining up to 3.4 ± 0.2 g L<sup>−1</sup> final isopropanol concentration with a specific yield of 1.4 ± 0.1 g g<sub>CDW</sub><sup>−1</sup>. Although the use of CO<sub>2</sub> instead of fructose resulted in a slower process, the overall isopropanol production is promising. This study provides valuable insights into strain behaviour while demonstrating the utility of the presented shake bottle system for advancing autotrophic process development.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"403 ","pages":"Pages 1-8"},"PeriodicalIF":4.1,"publicationDate":"2025-03-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143691831","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

Ethanolysis of degummed soybean oil using magnetic CLEAs from Eversa® Transform

IF 4.1 2区生物学

Journal of biotechnology Pub Date : 2025-03-20 DOI: 10.1016/j.jbiotec.2025.03.010

Letícia Passos Miranda , José Renato Guimarães , Roberto Fernandez-Lafuente , Paulo Waldir Tardioli

{"title":"Ethanolysis of degummed soybean oil using magnetic CLEAs from Eversa® Transform","authors":"Letícia Passos Miranda , José Renato Guimarães , Roberto Fernandez-Lafuente , Paulo Waldir Tardioli","doi":"10.1016/j.jbiotec.2025.03.010","DOIUrl":"10.1016/j.jbiotec.2025.03.010","url":null,"abstract":"<div><div>Eversa<sup>@</sup> Transform magnetic crosslinked enzyme aggregates (Eversa-mCLEA) have been used to produce fatty acid ethyl esters (FAEEs) through the ethanolysis of soybean oil. Some variables influencing this reaction were studied using an experimental statistical design. After 12 hours of reaction, a maximum FAEEs yield of 64 wt% was obtained using 4 U<sub>est</sub>/g oil of Eversa-mCLEA, an anhydrous ethanol/refined oil molar ratio of 11, and a temperature of 40°C. Degummed oil and hydrated ethanol were used as more cost-effective alternatives, leading to an increase in FAEEs yield (up to 73 wt%). The initial reaction rate increased with a lower molar ratio of hydrated ethanol/degummed oil; however, the final yield remained similar. The combined use of Eversa-mCLEA and Lipozyme 435 resulted in 86 wt% FAEEs and 4 wt% of free fatty acids (FFAs) after 24 hours. A caustic polishing step of the product yielded 90 wt% FAEEs and 0.17 wt% FFAs. These findings show that, using these substrates, a more effective purification step (such as fractional distillation) is required for the product to meet international standards for biodiesel commercialization.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"402 ","pages":"Pages 79-86"},"PeriodicalIF":4.1,"publicationDate":"2025-03-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143687925","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

An efficient lysate-based approach for biosynthesis of the pyrrolobenzodiazepine natural product tilimycin

IF 4.1 2区生物学

Journal of biotechnology Pub Date : 2025-03-20 DOI: 10.1016/j.jbiotec.2025.03.012

Monica R. MacDonald , Andrew M. Gulick

{"title":"An efficient lysate-based approach for biosynthesis of the pyrrolobenzodiazepine natural product tilimycin","authors":"Monica R. MacDonald , Andrew M. Gulick","doi":"10.1016/j.jbiotec.2025.03.012","DOIUrl":"10.1016/j.jbiotec.2025.03.012","url":null,"abstract":"<div><div>Many bacteria use nonribosomal peptide synthetases (NRPSs), a family of multidomain enzymes that produce peptide natural products using an assembly line strategy. One class of such compounds are pyrrolobenzodiazepines, which have DNA alkylating activity. One example is tilimycin, a compound produced by the human gut microbiota that plays a role in epithelial damage during antibiotic-associated dysbiosis. The production of analogs of these natural products may facilitate the discovery of novel bioactive molecules. However, the synthesis of these natural products typically requires significant resources and time to produce in sufficient amounts for microbial and biochemical testing. Biocatalysis offers an environmentally friendly approach, but enzyme yield and stability, particularly with large multidomain enzymes that are often used in natural product pathways, can limit biochemical reactions with purified protein. Here, we demonstrate a cell lysate-based method to synthesize the nonribosomal peptide natural product tilimycin directly from the substrates 3-hydroxyanthranilic acid and L-proline with lysates from <em>E. coli</em> cell lines that express the tilimycin biosynthetic proteins. We present our protocol optimization and scale-up to produce tilimycin in a fast, efficient manner.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"402 ","pages":"Pages 87-95"},"PeriodicalIF":4.1,"publicationDate":"2025-03-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143687924","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

Synergistic interaction of AaMYC2 and AaMYC2-LIKE enhances artemisinin production in Artemisia annua L.

IF 4.1 2区生物学

Journal of biotechnology Pub Date : 2025-03-18 DOI: 10.1016/j.jbiotec.2025.03.007

Ishfaq Majid Hurrah , Mohammad , Amit Kumar , Nazia Abbas

{"title":"Synergistic interaction of AaMYC2 and AaMYC2-LIKE enhances artemisinin production in Artemisia annua L.","authors":"Ishfaq Majid Hurrah , Mohammad , Amit Kumar , Nazia Abbas","doi":"10.1016/j.jbiotec.2025.03.007","DOIUrl":"10.1016/j.jbiotec.2025.03.007","url":null,"abstract":"<div><div>Artemisinin-based combination therapies recommended by WHO marks <em>Artemisia annua</em> as the only natural source of artemisinin fighting deadly disease, Malaria. In this study, we isolated two transcription factors, AaMYC2 and AaMYC2-LIKE, from <em>A. annua</em> and investigated their role in regulating artemisinin biosynthetic pathway. Our findings depict that both AaMYC2 and AaMYC2-LIKE are transcriptionally active and, when co-transformed in yeast cells, significantly enhance β-galactosidase activity in transactivation assays as compared to their individual transformations. Furthermore, Yeast two-hybrid (Y2H) and Biomolecular fluorescence complementation assays revealed AaMYC2 physically interacts with AaMYC2-LIKE in yeast cells and in the nucleus of onion epidermal cells respectively. Generation of transient transgenic over expression and co-expression lines of AaMYC2 and AaMYC2-LIKE resulted in elevated expression of artemisinin biosynthetic genes and trichome development genes in co-expression lines as compared to individual transgenic lines and wildtype. Importantly, the glandular trichome density and artemisinin content was also significantly higher in co-transformed transgenic lines compared to individual AaMYC2 and AMYC2-LIKE transgenic lines. Conversely, artemisinin content was markedly reduced in AaMYC2-RNAi lines, underscoring the critical role of functional AaMYC2 in synergistic regulation with AaMYC2-LIKE. Altogether the above studies provide valuable insights into the regulatory networks of MYC type bHLH transcription factors in controlling economically and medically important pathway in <em>A. annua.</em></div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"402 ","pages":"Pages 69-78"},"PeriodicalIF":4.1,"publicationDate":"2025-03-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143663424","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