Journal of biotechnology最新文献

{"title":"Differential metabolic reprogramming due to nitrogen starvation leads to remobilization of storage metabolites in microalgal strains","authors":"Lidiane Covell ,&nbsp;Mariana Machado ,&nbsp;Marcelo Gomes Marçal Vieira Vaz ,&nbsp;Auxiliadora Oliveira Martins ,&nbsp;Carolina Cassano Monte Bello ,&nbsp;Umarah Mubeen ,&nbsp;Dilson Novais Rocha ,&nbsp;Marcio Arêdes Martins ,&nbsp;Camila Caldana ,&nbsp;Wagner L. Araújo ,&nbsp;Adriano Nunes-Nesi","doi":"10.1016/j.jbiotec.2025.11.008","DOIUrl":"10.1016/j.jbiotec.2025.11.008","url":null,"abstract":"<div><div>Microalgal stress-related responses to nitrogen (N) deprivation are determined by genetically defined metabolic changes, which are further influenced by cultivation and nutritional conditions. In this study, we evaluated the metabolic responses of five chlorophyte strains to varying N concentrations. The strains included <em>Chlamydomonas reinhardtii</em> and <em>Scenedesmus obliquus</em>, which accumulated higher levels of starch, and <em>Monoraphidium</em> sp., <em>Chlamydomonas</em> sp., and <em>Chlorella vulgaris</em>, characterized by differences in both the levels and compositions of accumulated lipids. Under N-starved condition, all strains exhibited increased starch and neutral lipid content, with strain-specific variations. In this treatment, the content of branched-chain amino acids decreased in <em>Monoraphidium</em> sp., likely due to amino acid catabolism associated with the autophagy and triacylglycerol (TAG) accumulation. Furthermore, we observed a remodeling of lipid classes in <em>S. obliquus</em>, <em>C. vulgaris</em>, <em>Chlamydomonas</em> sp., and <em>Monoraphidium</em> sp., including reductions in digalactosyl diacylglycerol, monogalactosyl diacylglycerol, and sulfoquinovosyl diacylglycerol, accompanied by an increase in TAGs. Under N-starvation several fatty acids of <em>C. vulgaris</em>, <em>Chlamydomonas</em> sp., and <em>C reinhardtii</em>, including α-linolenic acid (C18:3), oleic acid (C18:1), and linoleic acid (C18:2) increase. In contrast, <em>Monoraphidium</em> sp. accumulated long-chain fatty acids such as lumequic acid (C30:1), nervonic acid (C24:1), docosanoic acid (C22:0), and erucic acid (C22:1). Collectively, these results indicate that chlorphyte strains exhibit distinct metabolic responses to N depletion in the medium. These findings highlight the importance of strain-specific assessments when developing the N management strategies for microalgal cultivation and biotechnological applications.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"409 ","pages":"Pages 231-246"},"PeriodicalIF":3.9,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145482251","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}

{"title":"Increased purity and refolding yield of bacterial inclusion bodies by recursive high pressure homogenization","authors":"Robert Klausser ,&nbsp;Lukas Veiter ,&nbsp;Julian Kopp ,&nbsp;Nikolaus Hammerschmidt ,&nbsp;Tiffany Frierss ,&nbsp;Florian Gisperg ,&nbsp;Mohamed Elshazly ,&nbsp;Eva Prada Brichtova ,&nbsp;Michael Martinetz ,&nbsp;Martin Voigtmann ,&nbsp;Oliver Spadiut","doi":"10.1016/j.jbiotec.2025.11.004","DOIUrl":"10.1016/j.jbiotec.2025.11.004","url":null,"abstract":"<div><div>Therapeutic proteins are often produced intracellularly in the bacterium <em>Escherichia coli</em> in aggregated form, known as inclusion bodies, which must be solubilized and refolded to recover the biologically active product. However, the industrially established refolding method of batch dilution is still carried out at low protein concentrations, leading to high water consumption, large equipment footprints, and poor sustainability. An underexplored way to increase refolding yields and thereby enable higher product concentrations after refolding is the optimization of cell lysis and inclusion body washing. In this study, we developed and evaluated a “recursive high pressure homogenization” approach, wherein wash steps and additional homogenization cycles were combined to enhance the final purity of inclusion bodies before solubilization. The degree of cell lysis, nucleic acid release and final inclusion body purity were compared to conventional “linear washing”, where all homogenization cycles are completed prior to the washing of inclusion bodies. In total, seven process variations were compared and the resulting batches of inclusion bodies were solubilized and refolded to investigate the effect of early downstream processing on the product concentration in the final refolds. For the most efficient process variant, the recursive high pressure homogenization protocol led to the highest measured product concentration of 855 mg/L after refolding. Thus, the proposed recursive high pressure homogenization approach led to an estimated 18 % reduction of CO₂ footprint caused by urea, and increased the product yield per biomass from 5.17 g/kg to 7.84 g/kg compared to its linear wash counterpart — without introducing non-standard equipment or chemicals.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"409 ","pages":"Pages 183-194"},"PeriodicalIF":3.9,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145471041","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}

{"title":"Engineering of endogenous plasmids in probiotic Escherichia coli Nissle 1917 for autonomous accumulation of 5‑aminolevulinic acid","authors":"Dong Yu ,&nbsp;Lin Zhu ,&nbsp;Ya-Xue Ding ,&nbsp;Li-Jing Mao ,&nbsp;Min Xiong ,&nbsp;Xiao-Xuan Jin ,&nbsp;Yujie Ma ,&nbsp;Feng-Qing Wang ,&nbsp;Liang-Bin Xiong","doi":"10.1016/j.jbiotec.2025.11.006","DOIUrl":"10.1016/j.jbiotec.2025.11.006","url":null,"abstract":"<div><div>5-aminolevulinic acid (ALA), a non‑proteinogenic δ‑amino acid, is a versatile compound with applications as a tumor-sensitizing agent in photodynamic therapy and a plant biostimulant that enhances stress tolerance and photosynthetic efficiency. This study aimed to leverage the endogenous cryptic plasmids pMUT1 and pMUT2 of the probiotic <em>Escherichia coli</em> Nissle 1917 (EcN) to construct a self-sufficient strain with autonomous ALA biosynthesis capability, eliminating dependencies on exogenous plasmids or chemical inducers. A synthetic operon was designed on a re-engineered cryptic plasmid to express core genes of the C5 pathway (<em>gltX</em>, <em>hemA</em>, and <em>hemL</em>). Systematic evaluation demonstrated that overexpression of <em>gltX</em> alone resulted in minimal ALA accumulation (6.8 mg/L). In contrast, coordinated co-expression of <em>hemA</em> and <em>hemL</em> significantly increased ALA titers to 854.5 mg/L, highlighting their synergistic role in channeling carbon flux through the C5 pathway in EcN. Subsequent optimization of the <em>hemA‑hemL</em> cassette further elevated production to 925.1 mg/L. Notably, this enhanced ALA synthesis perturbed the cellular NADP⁺ /NADPH balance. To address this, we integrated <em>pos5</em>, encoding a NADP⁺‑dependent transhydrogenase, into the endogenous plasmid, enabling <em>in situ</em> NADPH regeneration and boosting ALA titers to 1306.8 mg/L. Scale up to a 5-L bioreactor with fed-batch cultivation and controlled glycerol feeding achieved an ALA titer of 3372.5 ± 162.7 mg/L at 108 h. This endogenous plasmid-centric approach establishes an inducer-free, antibiotic-independent microbial cell factory, positioning EcN as a universal platform for ALA production with potential in biomedical and agricultural applications.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"409 ","pages":"Pages 157-164"},"PeriodicalIF":3.9,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145471033","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}

{"title":"CRISPR/anti-CRISPR genome editing in Clostridium beijerinckii","authors":"Rémi Hocq ,&nbsp;Gwladys Chartier,&nbsp;Nicolas Lopes Ferreira,&nbsp;François Wasels","doi":"10.1016/j.jbiotec.2025.11.002","DOIUrl":"10.1016/j.jbiotec.2025.11.002","url":null,"abstract":"<div><div>The development of CRISPR technologies has revolutionized genome editing. However, in bacteria, CRISPR-based methods can be difficult to implement due to the cytotoxicity of CRISPR-associated proteins, which often impair or entirely prevent transformation. In this work, we combine inducible expression of classical CRISPR-Cas9 components with the anti-CRISPR protein AcrIIA4 from <em>Listeria monocytogenes</em> to tightly regulate Cas9 activity. Using this approach, we demonstrate efficient and iterative genome editing in the genetically recalcitrant <em>Clostridium beijerinckii</em> DSM 6423. While deletion of <em>upp</em> alone was not sufficient to render the strain sensitive to 5-fluorouracil, the additional deletion of a second gene involved in the uracil salvage pathway conferred resistance to the drug and validated our gene editing strategy. Collectively, our results show that CRISPR/anti-CRISPR systems can overcome a key limitation of CRISPR-based genome editing and may offer a broadly applicable strategy for engineering otherwise intractable bacterial species.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"409 ","pages":"Pages 165-169"},"PeriodicalIF":3.9,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145471016","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}

{"title":"Impact of humanization scaffold design on the functional activity of FGFR2-targeting chicken scFvs","authors":"Shyi-Chun Yii ,&nbsp;Tsai-Yu Lin ,&nbsp;Fu-Ling Chang ,&nbsp;Fu-Shan Jaw ,&nbsp;Keng-Chang Tsai ,&nbsp;Yun-Shih Lin ,&nbsp;Yu-Ching Lee","doi":"10.1016/j.jbiotec.2025.10.005","DOIUrl":"10.1016/j.jbiotec.2025.10.005","url":null,"abstract":"<div><div>Fibroblast growth factor receptor 2 (FGFR2) is frequently overexpressed in gastric cancer and represents a promising therapeutic target. We developed FGFR2-specific single-chain variable fragments (scFvs) using a chicken-derived immune library and a refined cell-based panning strategy incorporating FGFR2-knockdown cells for negative selection. The lead clone, scFv R21, exhibited high binding affinity and strong tumor-inhibitory effects in vitro. To enable therapeutic application, R21 was humanized using three distinct framework strategies: CDR grafting, light chain replacement, and structure-guided back-mutation. Structural modeling by AlphaFold3 indicated that the hR21-Bf variant preserved CDR conformation and overall stability. Despite lower expression, hR21-Bf retained moderate FGFR2 binding affinity (<em>K</em>_D) of 38 nM, while other variants showed markedly reduced reactivity. The hR21-Bf construct was further reformatted as a full-length human IgG1 and evaluated in a gastric cancer xenograft model. Mice treated with IgG hR21-Bf showed significant tumor growth inhibition without observable toxicity. Immunohistochemical and biochemical analyses of resected tumors confirmed reduced Ki-67 expression and downregulation of FGFR2-mediated signaling. Our study highlights the impact of scaffold selection on antibody structure and function, supporting a rational approach to antibody humanization using avian-derived libraries for cancer therapy.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"409 ","pages":"Pages 57-66"},"PeriodicalIF":3.9,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145329508","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}

{"title":"Efficient CRISPR/Cas-based gene editing in cotton induced by cotton leaf crumple virus","authors":"Jiajun Zhang ,&nbsp;Peihong Dai ,&nbsp;Zheng Weng ,&nbsp;Runze Xu ,&nbsp;Yue Li ,&nbsp;Xiaodong Liu ,&nbsp;Jianfeng Lei","doi":"10.1016/j.jbiotec.2025.10.001","DOIUrl":"10.1016/j.jbiotec.2025.10.001","url":null,"abstract":"<div><div>Plant viral vectors can replicate autonomously and spread within host cells, making them an ideal tool for the delivery of CRISPR/Cas gene-editing elements. Here, we constructed a cotton CRISPR/Cas system mediated by cotton leaf crumple virus (CLCrV) as a delivery vector. We first inoculated Pro35s::Cas9 and ProUbi::Cas9 cotton with sgRNAs designed to knock out <em>GhAGL16</em>, <em>GhPDS</em>, and <em>GhCLA1</em> target genes via the CLCrV vector and then compared the effects of these two transformation receptors on the editing efficiency of the same target genes. We next explored the feasibility of simultaneous multi-target editing in cotton via pooled virus inoculation. Finally, we used a cotton line overexpressing nCas9-TadA7.10 as the transformation receptor to explore the feasibility of CLCrV-mediated adenine base editing and verify the specificity of gene editing in this system. Mutation detection and deep sequencing revealed that the Pro35s::Cas9 and ProUbi::Cas9 cotton lines did not differ significantly in editing efficiency, and both could be used as successful receptors for the CLCrV-mediated Cas9 system. Pooled inoculation with CLCrV-sgRNAs enabled the simultaneous editing of multiple target genes in Pro35s::Cas9 and ProUbi::Cas9 cotton, although this approach had somewhat lower editing efficiency than inoculation with single sgRNAs. The CLCrV-mediated adenine base-editing system enabled A-to-G conversion at target sites in cotton <em>GhPEBP</em> and showed high gene-editing specificity. In summary, this study establishes an efficient CLCrV-mediated CRISPR system in cotton, providing a powerful technical tool for editing of multiple target genes and base editing.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"409 ","pages":"Pages 33-43"},"PeriodicalIF":3.9,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145274785","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}

{"title":"Bioconversion for ursodeoxycholic acid using β-cyclodextrin included-chenodeoxycholic acid via immobilized cells of Xanthomonas maltophilia","authors":"Zengyan Yang ,&nbsp;Caixia Liu ,&nbsp;Yan Jin ,&nbsp;Jing Wang ,&nbsp;Yingying Ma ,&nbsp;Juan Zou ,&nbsp;Yinlei Han ,&nbsp;Yixin Guan ,&nbsp;Wenting Zhao ,&nbsp;Qingxi Wu","doi":"10.1016/j.jbiotec.2025.10.004","DOIUrl":"10.1016/j.jbiotec.2025.10.004","url":null,"abstract":"<div><div>Ursodeoxycholic acid is a prevalent clinical drug widely used for hepatobiliary diseases. It offers the advantages of good efficacy, high sensitivity, low toxic side effects and multifunctionality. However, its poor water solubility and arduous biopreparation have always been obstacles. This work presents, for the first time, a continuous aerobic-biotransformation system for synthesizing ursodeoxycholic acid. The process utilizes immobilized <em>Xanthomonas maltophilia</em> cells with substrate chenodeoxycholic acid/<em>β</em>-cyclodextrin inclusion complexes. The designed inclusion complexes possessed favourable stability with solubility improved by 122 times comparing to the pure drug. Repeated batches transformation was carried out and reaction parameters like flow speed, fluid direction and filling volume were optimized. The results showed that <em>X. maltophilia</em> cells were successfully immobilized in the microspheres with typical shell-core structure and good abrasion resistance. The optimal reaction conditions were as follows: flow speed of 1.0 mL/min, fluid direction from bottom to top in a reverse orientation, and filling volume set to 1:1. The entire continuous reaction could be recycled in three consecutive batches with no decrease in the conversion of the products. This study provides a new strategy for the preparation of ursodeoxycholic acid via bioconversion, which is valuable to explore the aerobic-based strain for biotransformation.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"409 ","pages":"Pages 44-56"},"PeriodicalIF":3.9,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145318410","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}

{"title":"Impact of benzene-ethanol extractives on deep eutectic solvent-mediated lignin extraction from poplar and eucalyptus","authors":"Rongxuan Li,&nbsp;Shubin Wu","doi":"10.1016/j.jbiotec.2025.10.011","DOIUrl":"10.1016/j.jbiotec.2025.10.011","url":null,"abstract":"<div><div>Deep eutectic solvents (DES), an emerging class of green solvents, have demonstrated remarkable efficiency in the fractionation of lignocellulosic biomass. Benzene-ethanol extractives in woody biomass significantly affect the efficiency of DES-mediated fractionation. Therefore, investigating influence of benzene-ethanol extractives on DES-mediated pre-treatment is essential for optimizing of poplar and eucalyptus fractionation. Removing benzene-ethanol extractives significantly increased the polar component of the surface free energy (SFE) in both poplar and eucalyptus biomass. This prominently enhanced the wettability of DES on the raw materials, thereby ameliorating heat and mass transfer effectiveness. Furthermore, the removal of these extractives was instrumental in modifying the surface pore structure of the raw materials, thereby facilitating a further meliorative fractionation effect of DES. Following the removal of the benzene-ethanol extract, the delignification efficiency for poplar and eucalyptus significantly increased to 81.92 % and 80.16 %, respectively. Concurrently, the extracted lignin exhibited higher hydroxyl content, a more complex aromatic ring structure, and improved thermal stability. All these findings substantiate that the removal of benzene-ethanol extractives represents a promising strategy for enhanced DES fractionation efficiency.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"409 ","pages":"Pages 107-116"},"PeriodicalIF":3.9,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145400799","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}

{"title":"Hairy roots as a biotechnological tool for medicinal plant secondary metabolites: A systematic review","authors":"Amine Elbouzidi ,&nbsp;Mohamed Taibi ,&nbsp;Mounir Haddou ,&nbsp;Abdellah Baraich ,&nbsp;Ibrahim Sadougui ,&nbsp;Reda Bellaouchi ,&nbsp;Abdeslam Asehraou ,&nbsp;Khalid Chaabane ,&nbsp;Bouchra El Guerrouj ,&nbsp;Mohamed Addi","doi":"10.1016/j.jbiotec.2025.10.010","DOIUrl":"10.1016/j.jbiotec.2025.10.010","url":null,"abstract":"<div><div>Hairy root cultures, generated via Agrobacterium rhizogenes-mediated transformation, are a major advance in plant biotechnology. They exhibit genetic stability, autonomous growth without exogenous phytohormones, and sustained high-yield production of bioactive secondary metabolites. These systems have applications in pharmaceuticals, nutraceuticals, and cosmetics. Notable metabolites include vincristine from <em>Catharanthus roseus</em> (L.) G. Don, withanolides from <em>Withania somnifera</em> (L.) Dunal, and ginsenosides from <em>Panax ginseng</em> C.A. Meyer, displaying anticancer, anti-inflammatory, antimicrobial, and neuroprotective activities. Despite these advantages, challenges such as suboptimal yields in certain species, inefficient transformation protocols, and complex regulatory frameworks limit industrial adoption. To address this, a systematic review was performed following PRISMA guidelines. Data were retrieved from PubMed, Scopus, and Web of Science using terms including “hairy root culture,” “secondary metabolite biosynthesis,” and “elicitation strategies.” The review included experimental studies on medicinal plant species capable of metabolite production via hairy roots, excluding theoretical studies and non-medicinal plants. The review pursued five primary objectives: (1) to compile a comprehensive inventory of medicinal plant species utilized in hairy root research; (2) to critically evaluate elicitation methods for enhancing metabolite production; (3) to examine current challenges related to scale-up of hairy root cultures for industrial application; (4) to identify actionable strategies to overcome existing limitations; and (5) to highlight the pharmaceutical properties of secondary metabolites derived from hairy roots. Empirical findings indicate that elicitors—such as jasmonic acid, salicylic acid, and emerging nanomaterials—significantly enhance metabolite accumulation. Molecular tools further optimize biosynthetic pathways. Nevertheless, species-specific constraints and unharmonized regulatory guidelines continue to impede commercialization. Integration of genetic engineering, bioprocess optimization, and regulatory science is essential to fully exploit the biotechnological potential of hairy root culture systems for pharmaceutical development.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"409 ","pages":"Pages 117-136"},"PeriodicalIF":3.9,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145426441","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}

{"title":"Construction of high protein-producing mutant yeast strains via point and structural mutageneses and their use for carotenoid production","authors":"Ryosuke Yamada ,&nbsp;Yoshifumi Inoue,&nbsp;Yukino Karitani,&nbsp;Rumi Sakaguchi,&nbsp;Takuya Matsumoto,&nbsp;Hiroyasu Ogino","doi":"10.1016/j.jbiotec.2025.10.008","DOIUrl":"10.1016/j.jbiotec.2025.10.008","url":null,"abstract":"<div><div>The yeast <em>Saccharomyces cerevisiae</em> is a safe microorganism with established industrial-scale culture techniques. Standard laboratory <em>S. cerevisiae</em> strains, such as the representative YPH499, are valuable hosts for producing proteins and chemicals through metabolic engineering. Consequently, there's a high demand for platform strains of <em>S. cerevisiae</em> with enhanced protein production capacity. We have previously established an efficient and straightforward technique for introducing point and structural mutations into yeast via plasmid introduction, leading to the generation of mutant strains with superior phenotypes. In this study, we aimed to develop <em>S. cerevisiae</em> mutants with high protein production capacity using techniques to introduce point and structural mutations. We introduced these mutations into the YPH499/pEUPGGFP strain, which expresses green fluorescent protein (GFP). Since GFP is easily detected by its fluorescence, we selected mutants based on their fluorescence intensity. Consequently, YPH499/pEUPGGFP/Mu10G39, with a GFP fluorescence intensity 2.5-fold higher than that of the parent strain, was successfully obtained. Then, a carotenoid-producing plasmid was introduced to construct YPH499Mu10G39/pEU20Beta3. YPH499Mu10G39/pEU20Beta3 produced 6.74 mg/g-dry cell carotenoids, which was 2.9-fold higher than that produced by the parent strain. Transcriptome analysis suggested that YPH499Mu10G39 exhibited improved energy production, amino acid precursor supply, ribosome function, and stress tolerance, which may have contributed to its high protein production. In conclusion, by introducing point and structural mutations, we successfully developed the platform strain, YPH499Mu10G39, which is useful for the high production of various proteins. In the future, various proteins and useful chemicals can be produced through metabolic engineering using YPH499Mu10G39 as a platform strain.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"409 ","pages":"Pages 77-84"},"PeriodicalIF":3.9,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145359795","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}