Journal of biotechnology最新文献

{"title":"Integrative omics approaches for bioactive metabolite discovery in marine macroalgae: Recent advances and future perspectives","authors":"M. Marimuthu ,&nbsp;M. Anitha ,&nbsp;P. Prakash ,&nbsp;Meivelu Moovendhan","doi":"10.1016/j.jbiotec.2026.01.007","DOIUrl":"10.1016/j.jbiotec.2026.01.007","url":null,"abstract":"<div><div>Phlorotannins, bromophenols, sulfated polysaccharides, terpenoids, lipids and halogenated molecules exhibit potent antioxidant, anticancer, anti-inflammatory, and antimicrobial properties. Conventional discovery methods, such as solvent extraction and bioassay-guided fractionation, are limited by low throughput, specificity, and poor insight into biosynthetic mechanisms. High-throughput omics platforms including genomics, transcriptomics, proteomics, and metabolomics have transformed the discovery of bioactive metabolites in macroalgae by unraveling system-wide interpretation of biosynthetic pathways. This review outlines omics-driven strategies for macroalgal natural product research, highlighting genome sequencing and annotation efforts. Transcriptome-based biosynthetic gene cluster (BGC) mining, and gene regulation analysis via RNA-seq. Proteomic approaches such as 2D electrophoresis, LC-MS/MS, and MALDI-TOF are discussed for their role in elucidating enzyme functions and post-translational modifications. Metabolomics tools, including GC-MS, LC-MS, and NMR, paired with platforms like GNPS and MetaboAnalyst, have improved metabolite identification, dereplication and pathway analysis. Emerging integrative multi-omics frameworks combining machine learning and systems biology are paving the way for predictive, precision-driven bioprospecting. Nonetheless, challenges persist, including incomplete genome assemblies, taxonomic inconsistencies and limited reference databases. This review emphasizes how omics technologies are deciphering and mechanistically elucidating the untapped metabolic potential of marine macroalgae, accelerating their application in biotechnology and therapeutic innovation.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"411 ","pages":"Pages 13-28"},"PeriodicalIF":3.9,"publicationDate":"2026-01-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146018557","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":"Integrative experimental and computational approach for the development of novel plasminogen activators from truncated streptokinase fragments","authors":"Vijay Gunasekaran ,&nbsp;Suthakaran Pichaimuthu ,&nbsp;Vigneshwaran Namasivayam ,&nbsp;V. Ponnusami","doi":"10.1016/j.jbiotec.2026.01.004","DOIUrl":"10.1016/j.jbiotec.2026.01.004","url":null,"abstract":"<div><div>Streptokinase (SK), a widely used thrombolytic agent, activates human plasminogen, which in turn dissolves blood clots. However, its clinical application is limited because it elicits immunogenic responses in patients. The present study aims to develop novel plasminogen activators derived from truncated fragments of SK that retain activity comparable to the full-length SK enzyme. Based on the structure-function relationships and the previous literature, ten fragments of SK were designed by systematically truncating the N- and C-terminal regions and the central β-domain. These fragments included variants lacking N-terminal residues (amino acids 1–24 or 1–37), C-terminal truncations (beyond amino acid 300) and segments focused on the central domain (starting at amino acid 130 or ending at 173). Fragments were expressed in <em>E. coli</em> and their plasminogen activation activity was evaluated. The experimental results indicated that fragments containing an intact N1 domain retained a high level of plasminogen activation activity (fragments 1–300, 1–173), while deletion of key N-terminal residues significantly reduced it. C-terminal fragments partially compensated for N-terminal loss (fragment 130–414), but overall activity remained lower. Molecular modelling studies of protein–protein complex interface analyses revealed that the N1 domain (residues 1–173) of SK constitutes the principal binding interface with HPlg, with specific N-terminal residues serving as interaction hotspots. Residue-level contact mapping correlated strongly with experimental activity, underscoring the importance of precise interface residues rather than overall domain-level contact coverage. The study identifies SK fragments 1–300, 1–173 and 130–414 as having substantial activity comparable to full-length SK, suggesting that these constructs could serve as the basis for novel thrombolytic drug candidates pending further immunogenicity evaluation. These interesting findings provide both structural and functional insights for the rational design of improved streptokinase-derived therapeutics.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"411 ","pages":"Pages 29-39"},"PeriodicalIF":3.9,"publicationDate":"2026-01-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145997740","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":"Couple action of some proteases from feather keratin-degrading bacterium B.licheniformis CP-16","authors":"Shijin Luo ,&nbsp;Deshan Wang ,&nbsp;Kaige Yang ,&nbsp;Hui Ye ,&nbsp;Yingxia Sun ,&nbsp;Zhiguo Qi ,&nbsp;Jianjun Zuo ,&nbsp;Tieying Zhang","doi":"10.1016/j.jbiotec.2026.01.006","DOIUrl":"10.1016/j.jbiotec.2026.01.006","url":null,"abstract":"<div><div>A keratin-degrading bacterium, <em>Bacillus licheniformis</em> CP-16 (<em>B.licheniformis</em> CP-16), isolated from feather waste, was found to produce three extracellular proteins with protease activity. Following mass spectrometry sequencing and BLAST analysis, these proteins exhibited high similarity to γ-glutamyl transpeptidase, alkaline serine protease and thioredoxin-like protein YkuU, respectively. Genes of the three proteins and Ker A from <em>B.licheniformis</em> were successfully cloned into pET22b and expressed in <em>Escherichia coli</em> (<em>E coli</em>), and the four recombinant enzymes were named P-Glu, P-Alk, P-Trx, and P-Ker. P-ker showed great keratinase activity (4.9 kU/mg) and casein protease activity (6.5 kU/mg), as P-Alk showed keratinase activity (1.2 kU/mg) and casein protease activity (23 kU/mg). In contrast, both P-Glu and P-Trx displayed low levels of keratinase and casein protease activities. P-Trx had disulfide bond reductase activity (3 U/mg). When mixing with P-Trx, the other three proteases, P-Glu, P-Alk, P-Ker degraded feather keratin with keratinase activity promotion of 71 %, 40 % and 71 %. Commercial proteases (trypsin and chymotrypsin) also got keratinolytic capacity after mixing with P-Trx. These findings reveal a novel synergistic mechanism between P-Trx and other proteases in the degradation of feather keratin, leading to more efficient keratin breakdown.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"411 ","pages":"Pages 130-143"},"PeriodicalIF":3.9,"publicationDate":"2026-01-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145989140","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":"Marine-derived Streptomyces sp. isolated from the Persian Gulf as a novel source of melanin","authors":"Seyed Muhammad Hamid Malekpour ,&nbsp;Moj Khaleghi ,&nbsp;Alireza Akhtarpoor ,&nbsp;Mostafa Pournamdari ,&nbsp;Hoda KeshmiriNeghab ,&nbsp;Farideh MohammadHosseinZadeh","doi":"10.1016/j.jbiotec.2026.01.005","DOIUrl":"10.1016/j.jbiotec.2026.01.005","url":null,"abstract":"<div><div>In this study, the structure and biological properties of melanin pigment produced by Streptomyces strain SEA5 isolated from Persian Gulf sediments were investigated. This strain was cultured in an enriched culture medium (<span>L</span>-tyrosine, meat extract). Pigment production was also performed in culture media containing different sources of amino acid <span>L</span>-tyrosine such as soybean meal. Pigment extraction was performed using hydrochloric acid (6 M). In order to investigate its physicochemical nature, TLC, HPLC, UV-Vis spectroscopy, FTIR and 1 H NMR analysis was performed. On the other hand, strong UV absorption with SPF, antioxidant activity and cytotoxicity assay of this pigment were also studied. The results showed that strain SEA5 was well grown and melanized well (1.5 g/L) in medium containing <span>L</span>-tyrosine. It was 1.36 g/L when cultured in medium containing soybean meal. The UV–vis spectrum had an absorption peak at 225 nm. FTIR identified the functional groups of eumelanin with very high peaks at 3414.46 cm-1 (N-H group) and 1650–1540 cm-1 (C<img>O and C<img>N vibrations). Also, the ¹HNMR spectrum showed several peaks between 6.9 and 8.0 ppm, indicating aromatic protons. On the other hand, the extracted pigment had an SPF of 22.55 and showed significant antioxidant activity with 60–77 % reduction of DPPH radicals. The pigment in the formulation selectively inhibited the growth of A375 melanoma cells (IC50- 79.82 μg/mL), whilst it was non-toxic when tested against normal human dermal fibroblasts and HaCaT keratinocytes. Overall, these findings suggest the potential of SEA5-derived melanin as a human skin care product, pharmaceutical product and antioxidant material.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"411 ","pages":"Pages 78-88"},"PeriodicalIF":3.9,"publicationDate":"2026-01-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145989190","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":"Corrigendum to “Structure–function analysis and exonuclease deletion yield an improved strand-displacing DNA polymerase from Aeribacillus pallidus for efficient recombinase polymerase amplification” [J. Biotechnol. 410 (2026) 321–330]","authors":"Koki Nishi ,&nbsp;Eisuke Inoue ,&nbsp;Yuto Murakami ,&nbsp;Yuri Ishii ,&nbsp;Yukiko Nakura ,&nbsp;Itaru Yanagihara ,&nbsp;Kiyoshi Yasukawa ,&nbsp;Shinsuke Fujiwara","doi":"10.1016/j.jbiotec.2026.01.003","DOIUrl":"10.1016/j.jbiotec.2026.01.003","url":null,"abstract":"","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"411 ","pages":"Page 12"},"PeriodicalIF":3.9,"publicationDate":"2026-01-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145981351","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":"Two industrial media reveal a mitochondrial disfunction in CHO cell cultures co-fed with glucose and lactic acid","authors":"Keegan Orzechowski ,&nbsp;Johanna Vappiani-Korben ,&nbsp;Daniel C. Sevin ,&nbsp;Juan Aon","doi":"10.1016/j.jbiotec.2026.01.002","DOIUrl":"10.1016/j.jbiotec.2026.01.002","url":null,"abstract":"<div><div>Metabolomics analyses of cell culture processes can provide valuable insight into cellular physiology that can be leveraged to develop more productive processes. In this work, we applied metabolomics to interrogate CHO cell behavior in two industrial chemically-defined media in cultures co-fed with glucose and lactic acid. We previously reported that secreted acylcarnitines are indicative of altered mitochondrial metabolism when cultures are fed lactic acid and serve to maintain homeostasis between free CoA, acetyl-CoA, free carnitines, and acylcarnitines (Vappiani <em>et al</em>., 2021). One of the two media (“Medium B”) increased significantly viable-cell count and antibody titer than Medium A. Here, we report that CHO’s mitochondrial dysfunctionality based on the secretion of acylcarnitines in lactic acid-fed cultures depends on the overall medium composition. We hypothesize that in order to achieve better growth and titer, Medium B exhibited an increased oxidative phosphorylation based on the lower secretion of acylcarnitines and a differential utilization of riboflavin and thiamine, precursors of coenzymes required to enhance mitochondrial pyruvate incorporation and TCA cycle function. Therefore, our data provides further evidence that non-obvious changes to medium composition can have substantial effects on CHO-based production processes by altering the activity of oxidative phosphorylation required for the proper functioning of mitochondria but also for better antibody production.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"411 ","pages":"Pages 1-11"},"PeriodicalIF":3.9,"publicationDate":"2026-01-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145933438","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-based precise methylation of specific FUT8 promoter regions allows isolation of CHO cells with a fine-tuned glycoprofile","authors":"Víctor Jiménez Lancho ,&nbsp;Klaus Leitner ,&nbsp;Kitty Agarwal ,&nbsp;Arathi Krishnakumar ,&nbsp;Anurag Khetan ,&nbsp;Nicole Borth ,&nbsp;Nicolas Marx","doi":"10.1016/j.jbiotec.2026.01.001","DOIUrl":"10.1016/j.jbiotec.2026.01.001","url":null,"abstract":"&lt;div&gt;&lt;div&gt;A major advantage of producing therapeutic proteins in mammalian cells is their ability to tailor proteins with human-like posttranslational modifications such as glycosylation, which ultimately defines aspects like stability, protein folding or immunogenicity. However, producing therapeutic proteins with a consistent and reproducible glycoprofile remains a major challenge for the biopharmaceutical industry, especially with biosimilar production. While the enzymes responsible for glycosylation of proteins have been the subject of various cell engineering approaches, tuning their gene expression to precise levels is still difficult to achieve. While CRISPR/Cas9 enabled the genetic engineering of cells to drastically overexpress or remove a target gene, CRISPR/dCas9-based epigenetic editing by targeted DNA methylation promises to stably change the expression pattern of target genes after transient transfection of the CRISPR-tool. Application of targeted DNA methylation so far has mostly been used to completely silence gene expression by fully methylating the corresponding promoter regions. Here, we aim to tune expression of the associated gene by DNA methylation of confined promoter regions and to apply this technique as a new glycoengineering approach. By coupling CRISPR-based targeted DNA methylation with lectin-FACS assisted sorting we obtained CHO cell lines with a fine-tuned phenotype. First, dCas9-DNMT3A3L in combination with one single gRNA is targeted to the FUT8 promoter to induce confined DNA methylation, resulting in a phenotypically diversified population. Next, a window sorting strategy based on lectin-stained cells using five different sorting gates spanning from low to high FUT8 expression was applied to isolate single clones with a defined phenotype. Isolated clones were phenotypically assessed and re-sorted to obtain a homogenous expression profile. The resulting clonal cell lines showed either tuned or knock-down phenotypes with varying gene expression levels. Two out of seven clones that showed tuned FUT8 gene expression were phenotypically stable over 60 days. Gene expression levels, on the other hand, showed a steady decline over time that in part, however, can be explained by the general variation of FUT8 expression in different growth phases. Importantly, glycan analysis of recombinant EpoFc produced in the tuned clonal cell lines showed ranges of 35–70 % fucosylation, demonstrating that isolated clones can produce recombinant proteins with a distinct glycosylation profile. To understand why some clones showed tuned FUT8 gene expression levels while others were completely knocked-down, we analyzed the DNA methylation status of their respective FUT8 promoter. Critical areas within the FUT8 promoter were identified, with some associated with general repression and others with the tuning of FUT8 gene expression when affected by DNA methylation. Additionally, a combination of histone marks associated with active and rep","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"410 ","pages":"Pages 341-352"},"PeriodicalIF":3.9,"publicationDate":"2026-01-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145933379","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":"Structure–function analysis and exonuclease deletion yield an improved strand-displacing DNA polymerase from Aeribacillus pallidus for efficient recombinase polymerase amplification","authors":"Koki Nishi ,&nbsp;Eisuke Inoue ,&nbsp;Yuto Murakami ,&nbsp;Yuri Ishii ,&nbsp;Yukiko Nakura ,&nbsp;Itaru Yanagihara ,&nbsp;Kiyoshi Yasukawa ,&nbsp;Shinsuke Fujiwara","doi":"10.1016/j.jbiotec.2025.12.020","DOIUrl":"10.1016/j.jbiotec.2025.12.020","url":null,"abstract":"<div><div>Isothermal nucleic-acid amplification relies on strand-displacing DNA polymerases that synthesize DNA while unwinding duplex templates. We previously identified two thermostable family-A polymerases, C1-Pol from <em>Geobacillus zalihae</em> and H1-Pol from <em>Aeribacillus pallidus</em>, which show a characteristic and reproducible trade-off between stability and activity. C1-Pol is more thermostable, whereas H1-Pol exhibits stronger strand-displacement activity and higher recombinase polymerase amplification (RPA) efficiency. Domain-swapping analysis indicated that a chimeric construct (C1-Pol#4), in which the C1-Pol 5′→3′ exonuclease region was replaced with that of H1-Pol, enhanced strand-displacement and nucleotide-incorporation activities while maintaining parental-level stability, suggesting that this region modulates these important biochemical properties. However, C1-Pol#4 did not improve RPA efficiency. To clarify the functional contribution of this domain, we constructed Klenow-type deletion mutants lacking the 5′→3′ exonuclease region. The H1-Pol large fragment (H1-PolLF) exhibited reduced thermostability but markedly enhanced polymerase and strand-displacement activities at 40 °C, the optimal temperature for RPA reactions. At this temperature, H1-PolLF showed ∼10-fold higher strand-displacement activity than Bst DNA polymerase 2.0 and 4.4-fold higher than full-length H1-Pol. H1-PolLF also accelerated RPA, producing equivalent amplicons in half the reaction time and improving detection sensitivity 100-fold (6 × 10³ → 6 × 10¹ copies) than parental H1-Pol. Coupling H1-PolLF with branched-chain polyamine (BCPA)–conjugated magnetic beads enabled reliable detection of ∼10² target molecules from 10 mL saline. These findings demonstrate that removing the 5′→3′ exonuclease domain fine-tunes polymerase and strand-displacing functions, yielding an enzyme highly suited for rapid, ultrasensitive isothermal nucleic-acid detection when used in combination with BCPA beads.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"410 ","pages":"Pages 321-330"},"PeriodicalIF":3.9,"publicationDate":"2026-01-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145900250","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":"Engineered Lactococcus lactis expressing antimicrobial peptide HI: Enhanced survival and protection against ETEC in mice","authors":"Mingyang Hu ,&nbsp;Chongpeng Bi ,&nbsp;Yuwen Li,&nbsp;Yutong Xue,&nbsp;Sina Cha,&nbsp;Lu Zhao,&nbsp;Chenyu Xue,&nbsp;Na Dong","doi":"10.1016/j.jbiotec.2025.12.019","DOIUrl":"10.1016/j.jbiotec.2025.12.019","url":null,"abstract":"<div><div>The rising prevalence of antibiotic resistance underscores the urgent need for alternative strategies to manage pathogenic bacteria. Engineered probiotics offer a promising platform for delivering antimicrobial peptides (AMPs); however, their practical application remains constrained by challenges related to maintaining viability and <em>in vivo</em> functionality. This study focused on two main aspects: (1) optimizing a freeze-drying strategy for <em>Lactococcus lactis/</em>pNZC-Usp45-H-6 ×His (<em>L. L</em>/HI), which expresses the AMP HI targeting <em>Escherichia coli</em>, and (2) evaluating its protective efficacy against enterotoxigenic <em>Escherichia coli</em> (ETEC) infection in a murine model. Sorbitol at a concentration of 6 % (w/v) was identified as the most effective cryoprotectant for preserving bacterial viability after freeze-drying. In the ETEC infection model, oral administration of <em>L. L</em>/HI significantly alleviated intestinal injury by reducing bacterial colonization and lipopolysaccharide levels, alleviating inflammation, and restoring the expression of tight junction genes. Moreover, <em>L. L</em>/HI downregulated the expression of pro-inflammatory cytokines (TNF-α, IL-1β, IL-6) and upregulated the anti-inflammatory cytokine IL-10 in ileal tissues. These findings demonstrate that oral administration of <em>L. L</em>/HI reduced the bacterial burden in the ileum of ETEC–infected mice and indirectly alleviated inflammation and intestinal barrier damage caused by ETEC infection. This study provides a novel approach for the translational application of engineered probiotics.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"410 ","pages":"Pages 331-340"},"PeriodicalIF":3.9,"publicationDate":"2026-01-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145900309","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 an E. coli cell factory for ergothioneine through SAM-cycle enhancement and pathway reconstruction","authors":"Xiaoyu Zhang ,&nbsp;Jiayue Guan ,&nbsp;Yanqi Yang ,&nbsp;Shangci Zuo ,&nbsp;Chang Liu ,&nbsp;Pengchao Wang","doi":"10.1016/j.jbiotec.2025.12.018","DOIUrl":"10.1016/j.jbiotec.2025.12.018","url":null,"abstract":"<div><div>Ergothioneine (EGT) is a rare sulfur-containing derivative of methionine with potent antioxidant, anti-inflammatory, and neuroprotective properties. Its unique bioactivities make it a promising ingredient for applications in functional foods, nutraceuticals, and cosmetics. Microbial fermentation offers a sustainable alternative to extraction from natural sources, yet challenges such as precursor limitations, cofactor imbalances, and pathway complexity continue to restrict industrial-scale production. In this study, we engineered <em>Escherichia coli</em> (<em>E. coli</em>) as a microbial chassis for efficient de novo synthesis of EGT. By co-expressing key enzymes derived from bacteria and fungi, enhancing cysteine biosynthesis, and improving methionine utilization, we addressed key bottlenecks in precursor supply. Furthermore, the introduction of a methylation cycle significantly improved the regeneration of S-adenosylmethionine (SAM), relieving cofactor limitations. These combined metabolic engineering strategies led to a substantial increase in EGT production. The final engineered strain achieved a titer of 141.3 mg/L in shake flasks, representing a sixfold improvement over the base strain. In a 5-liter fed-batch fermentation, the titer reached 1.95 g/L without precursor supplementation and further increased to 2.52 g/L upon low-dose amino acid feeding. This work establishes a cost-effective and scalable biosynthetic platform for EGT production in <em>E. coli</em>, offering a viable route for its application in food and health-related industries.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"410 ","pages":"Pages 298-308"},"PeriodicalIF":3.9,"publicationDate":"2025-12-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145878348","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}