Journal of biotechnology最新文献

{"title":"Leaf vein scaffolds for three-dimensional culture of PDLSCs-derived Muse cells","authors":"Xueting Bi ,&nbsp;Qian Lan ,&nbsp;Xin Xiao ,&nbsp;Yu Deng ,&nbsp;Dongsheng Li","doi":"10.1016/j.jbiotec.2025.06.005","DOIUrl":"10.1016/j.jbiotec.2025.06.005","url":null,"abstract":"<div><div>Periodontal disease, a significant global health burden, has encountered limited success with current therapeutic strategies to achieve full tissue regeneration. The emergence of Multilineage Differentiation and Stress-Enduring (Muse) cells presents a promising avenue for periodontal tissue regeneration. This study introduced a novel three-dimensional (3D) culture system utilizing Magnolia leaf vein scaffolds, characterized for their biocompatibility and evaluated for their impact on Muse cells' proliferation, adhesion, osteogenic differentiation, and exosome secretion. The isolation of Muse cells from Periodontal Ligament Stem Cells (PDLSCs) was successfully accomplished, with excellent compatibility observed with the plant-derived scaffolds. Notably, the 3D culture substantially upregulated osteogenic markers and promoted the formation of mineralized nodules, signifying enhanced osteogenic potential. Additionally, Muse cells in 3D culture exhibited a significant increase in exosome secretion, which were more effective in stimulating PDLSCs proliferation. The study concluded that plant leaf vein scaffolds provide a sustainable and effective platform for 3D stem cell culture, with the potential to significantly enhance the therapeutic efficacy of Muse cells in periodontal tissue engineering.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"405 ","pages":"Pages 275-282"},"PeriodicalIF":4.1,"publicationDate":"2025-06-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144279200","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":"A novel immobilized bioreactor based on CFD simulation for fuel ethanol production from corn","authors":"Qingguo Liu ,&nbsp;Jing Liu ,&nbsp;Alan Yan ,&nbsp;Wenjun Sun ,&nbsp;Caice Liang ,&nbsp;Tianpeng Chen ,&nbsp;Qingshi Wen ,&nbsp;Yanjun Chen ,&nbsp;Hanjie Ying ,&nbsp;Yong Chen","doi":"10.1016/j.jbiotec.2025.06.003","DOIUrl":"10.1016/j.jbiotec.2025.06.003","url":null,"abstract":"<div><div>This study evaluated the feasibility of ethanol production from corn in a surface immobilized bioreactor. The mass transfer of 50 L immobilized bioreactor was analyzed based on computational fluid dynamics (CFD) simulation. Compared to the traditional stirred fermenter, the surface immobilized fermenter system exhibited a relatively weak liquid velocity, but its flow field distribution was relatively uniform, which ensured an ideal interaction environment between cells and substrates. The difference of fermentation indexes between the two fermentation methods was then verified by ethanol fermentation using corn hydrolysate. It was found that the ethanol productivity (P) of immobilized cells was increased by 20.59 %, while the amount of corn consumed for one ton of ethanol was decreased by 5.90 %. To further enhance industrial application, the semi-continuous fermentation was adopted, the P and ethanol yield were increased by 30.88 % and 3.06 percentage points, respectively, compared to free-cell fermentation (FCF). This work demonstrated that the mass transfer issues associated with traditional immobilized-cell fermentation (ICF), such as embedding and chemical crosslinking, were resolved through bioreactor design and process optimization. Meanwhile, the surface immobilization technology showcased special advantages, including a broad spectrum of raw materials and strong process stability, indicating great industrial application potential in ethanol production.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"405 ","pages":"Pages 254-262"},"PeriodicalIF":4.1,"publicationDate":"2025-06-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144239563","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":"Brazilian Amazon red propolis as a sustainable resource for the green synthesis of antibacterial silver nanoparticles","authors":"Roberto Pereira Santos ,&nbsp;Jaqueline Daniele Santos Barros ,&nbsp;Euzinete Borges Pereira ,&nbsp;Karla Gabriela Mota de Oliveira ,&nbsp;Gabriel Sousa Brito ,&nbsp;Fernanda Farias Costa ,&nbsp;Queli Cristina Fidelis ,&nbsp;Aramys Silva Reis ,&nbsp;Carlos Alexandre Holanda ,&nbsp;Richard Pereira Dutra","doi":"10.1016/j.jbiotec.2025.05.018","DOIUrl":"10.1016/j.jbiotec.2025.05.018","url":null,"abstract":"<div><div>This study investigates the use of Amazon red propolis for the green synthesis of antibacterial silver nanoparticles (AgNPs). A hydroalcoholic propolis extract was fractionated to yield fractions rich in phenolic compounds. High-performance liquid chromatography identified calycosin as a chemical marker, along with liquiritigenin, isoliquiritigenin, and formononetin. The propolis extract and the chloroform fraction inhibited <em>Staphylococcus aureus</em> growth with a bactericidal effect, while for <em>Escherichia coli</em>, a bacteriostatic effect was observed at concentrations below 1000 µg/mL. The AgNPs were characterized by UV-Vis spectroscopy, confirming surface plasmon resonance, and their morphology and size were analyzed using dynamic light scattering and transmission electron microscopy, with elemental composition assessed via energy-dispersive spectroscopy. X-ray diffraction and selected area electron diffraction showed the crystalline structure. Fourier-transform infrared spectra indicated that carboxyl and hydroxyl groups in flavonoids reduce silver ions and contribute to the nanoparticles' chemical stability. The nanoparticles synthesized from the propolis extract showed enhanced antibacterial potential, with efficacy against <em>E. coli</em> (MIC 25 µg/mL, MBC 50 µg/mL) compared to <em>S. aureus</em> (MIC 100 µg/mL, MBC 100 µg/mL), and demonstrated selectivity for Gram-negative bacteria over murine macrophages (CC₅₀ 83.92 μg/mL). These nanoparticles exhibited maximum absorption in the UV-Vis region at 418 nm, with a spherical silver core encapsulated by compounds from propolis, hydrodynamic diameter of 252.47 ± 1.42 nm, a zeta potential of −51.80 ± 0.70 mV, and a polydispersity index of 0.218 ± 0.01, indicating high chemical stability. These findings underscore Amazon red propolis as a sustainable resource for developing selective and stable antibacterial agents.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"405 ","pages":"Pages 263-274"},"PeriodicalIF":4.1,"publicationDate":"2025-06-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144247998","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":"Cultivation of Nannochloropsis oceanica using food grade industrial side streams: Effect on growth and relative abundance of selected amino acids and unsaturated fatty acids","authors":"Emil Gundersen,&nbsp;Charlotte Jacobsen","doi":"10.1016/j.jbiotec.2025.06.006","DOIUrl":"10.1016/j.jbiotec.2025.06.006","url":null,"abstract":"<div><div>Developing more resource-efficient food production is needed to sustainably support a growing world population. Microalgae present a promising, future source of food ingredients, especially if cultivated using industrial side streams. In this study, <em>Nannochloropsis oceanica</em> was cultivated using food grade process water from industrial enzyme production as a source of inorganic nutrients. Effects on growth and the nutritional quality of produced biomass, with focus on selected fatty acids and amino acids, were investigated. One of the two tested side streams replaced up to 40% of a commercial nutrients supplement with no negative effect on either growth or the content of nutritionally important amino- and fatty acids. The other tested side stream hampered both growth and significantly reduced the content of omega-3 fatty acids in the biomass at only 20% replacement level. The diverging results are likely caused by differences in the macro- and micronutrient profile, but further investigation into the side streams composition is needed to unravel the exact cause. If optimized, the use of such side streams can help reduce both the resource input and cost of producing <em>N. oceanica</em> as a future, sustainable source of food ingredients.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"405 ","pages":"Pages 249-253"},"PeriodicalIF":4.1,"publicationDate":"2025-06-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144239407","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":"Deletion of bikaverin and fusarubin biosynthesis gene clusters via CRISPR/Cas9 system in Fusarium fujikuroi and its effect on GA3 biosynthesis","authors":"Lianggang Huang ,&nbsp;Yixin Song ,&nbsp;Ningning Li ,&nbsp;Jie Gao ,&nbsp;Bo Zhang ,&nbsp;Zhiqiang Liu ,&nbsp;Yuguo Zheng","doi":"10.1016/j.jbiotec.2025.06.004","DOIUrl":"10.1016/j.jbiotec.2025.06.004","url":null,"abstract":"<div><div>Gibberellic acid (GA3) is a critical plant hormone with significant agricultural applications, yet its production in <em>Fusarium fujikuroi</em> is constrained by competition for metabolic precursors, particularly acetyl-CoA, which is essential for GA3 biosynthesis. The genome of <em>F. fujikuroi</em> harbors numerous secondary metabolite biosynthetic gene clusters that divert acetyl-CoA away from the GA3 pathway, thereby limiting its yield. To address this challenge, we employed the CRISPR/Cas9 system to delete the bikaverin and fusarubin biosynthesis gene clusters, which are known to compete with GA3 biosynthesis for acetyl-CoA. This genetic intervention resulted in a substantial increase in GA3 production, with the ΔBIKΔFSR strain yielding 31.67 % more GA3 compared to the wild-type strain. Notably, the deletion of these gene clusters not only enhanced GA3 biosynthesis but also improved mycelial growth and carbon assimilation, as evidenced by increased consumption of reducing sugars during fermentation. We further employed qRT-PCR to assess comparative expression levels of genes associated with the glycohydrolysis, glycolysis, and the TCA pathway in engineered strain. Results indicated that removing by-product gene clusters enhances the glycohydrolase system, accelerating carbon assimilation. Given the presence of dozens of secondary metabolite biosynthetic gene clusters in the <em>F. fujikuroi</em> genome, the strategy reported here offers a promising avenue for further enhancing GA3 production by targeting additional non-essential gene clusters.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"405 ","pages":"Pages 229-237"},"PeriodicalIF":4.1,"publicationDate":"2025-06-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144222429","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":"Enabling quantification of 2′-fucosyllactose via ligand-dependent thermal stabilization of BoGT6a","authors":"Nayeon Kim ,&nbsp;Jonghyeok Shin ,&nbsp;Jun-Seob Kim ,&nbsp;Dae-Hyuk Kweon","doi":"10.1016/j.jbiotec.2025.06.002","DOIUrl":"10.1016/j.jbiotec.2025.06.002","url":null,"abstract":"<div><div>2′-Fucosyllactose (2′-FL) is a major component of Human Milk Oligosaccharides (HMOs) that plays a crucial role in developing the neonatal immune system and modulating gut microbiota. Due to its health benefits, 2′-FL has gained industrial importance as a key ingredient in probiotic products and functional foods. Although quantifying 2′-FL is crucial for its economical production and nutritional management, conventional methods require expensive equipment and skilled personnel, making high-throughput quantification challenging. In this study, we present a simple and cost-effective method for 2′-FL quantification by utilizing the thermal stability of BoGT6a, a glycosyltransferase derived from <em>Bacteroides ovatus</em> that specifically binds to 2′-FL. Initially, the binding of BoGT6a and 2′-FL was confirmed, and we demonstrated that 2′-FL-bound BoGT6a is protected from thermal stress. To achieve rapid detection of 2′-FL, we fused BoGT6a with the fluorescent protein mCherry, resulting in mCherry-BoGT6a, and investigated its thermal stability and fluorescence in response to varying 2′-FL concentrations. Finally, we developed a 2′-FL quantification device that measures protein precipitation with the change of electrical voltage. These results demonstrate the reliability and industrial applicability of BoGT6a-based 2′-FL quantification technology.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"405 ","pages":"Pages 283-289"},"PeriodicalIF":4.1,"publicationDate":"2025-06-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144234212","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 biosynthesis of ectoine in engineered Vibrio natriegens","authors":"Tianzhi Fang ,&nbsp;Mengkai Hu ,&nbsp;Dandan Li ,&nbsp;Kun Liu ,&nbsp;Yu Chen ,&nbsp;Zhenglian Xue ,&nbsp;Qingtao Liu ,&nbsp;Tianwen Wang","doi":"10.1016/j.jbiotec.2025.06.001","DOIUrl":"10.1016/j.jbiotec.2025.06.001","url":null,"abstract":"<div><div>The compatible solute ectoine, an amino acid derivative, was initially discovered to be a critical factor conferring high resistance to harsh environments in which the producer microbes subsist. Its diverse chemical and biomedical properties have made ectoine a star molecule, necessitating environmentally friendly and efficient preparation methods. In this study, we cloned a biosynthetic gene cluster from a strain screened from samples collected from a historic sea-salt plant. This gene cluster supports inducible heterogeneous ectoine synthesis controlled by the T7 promoter in the recombinant halophile host <em>Vibrio natriegens</em> Vmax × 2. Investigation of the dosage effect of each gene in the cluster on ectoine synthesis identified <em>ectC</em> as the most important gene. We performed <em>in silico</em> directed evolution and Pythia analysis to design <em>ectC</em> mutants and screen those that positively contribute to ectoine synthesis. We used a sequence-generating tool to design degenerated EctC coding sequences to ensure EctC levels in recombinant <em>V. natriegens</em>. Ectoine synthesis was validated in a 5-L stirring fermenter. A titer of 56.47 g/L ectoine was achieved in 28 h through feed batch fermentation. This study demonstrated the power of protein language model-guided metabolic enzyme design and codon adaptation index-based coding sequence selection in biosynthesis pathway engineering. It also offers an example of ectoine biosynthesis in the rising star halophile host <em>V. natriegens</em>, contributing significantly to microbiology and biotechnology.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"405 ","pages":"Pages 238-248"},"PeriodicalIF":4.1,"publicationDate":"2025-06-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144231662","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":"De novo production of prenylnaringenin compounds by a metabolically engineered Escherichia coli","authors":"Daniela Gomes ,&nbsp;Joana L. Rodrigues ,&nbsp;Nigel S. Scrutton ,&nbsp;Ligia R. Rodrigues","doi":"10.1016/j.jbiotec.2025.05.017","DOIUrl":"10.1016/j.jbiotec.2025.05.017","url":null,"abstract":"<div><div>Prenylnaringenin (PN) compounds, namely 8-prenylnaringenin (8-PN), 3’-prenylnaringenin (3’-PN), and 6-prenylnaringenin (6-PN), are reported to have several interesting bioactivities. This study aimed to validate a biosynthetic pathway for <em>de novo</em> production of PN in <em>Escherichia coli</em>. A previously optimized <em>E. coli</em> chassis capable of efficiently <em>de novo</em> producing naringenin was used to evaluate eleven prenyltransferases (PTs) for the production of PN compounds. As PT reaction requires dimethylallyl pyrophosphate (DMAPP) as extended substrate that has limited availability inside the cells, clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated protein 12a (Cas12a) (CRISPR-Cas12a) was used to construct ten boosted DMAPP-E<em>. coli</em> strains. All the PTs, in combination with the naringenin biosynthetic pathway, were tested in these strains. Experiments in 96-well deep well plates identified twelve strains capable of producing PN. <em>E. coli</em> M-PAR-121 with the integration of the 1-deoxy-D-xylulose-5-phosphate synthase (DXS) gene from <em>E. coli</em> (<em>Ec</em>DXS) into the <em>lacZ</em> locus of the genome (<em>E. coli</em> M-PAR-121:<em>Ec</em>DXS) expressing the soluble aromatic PT from <em>Streptomyces roseochromogenes</em> (CloQ) and the naringenin biosynthetic pathway was selected as the best producer strain. After optimizing the production media in shake flasks, 160.57 µM of 3’-PN, 4.4 µM of 6-PN, and 2.66 µM of 8-PN were obtained. The production was then evaluated at the bioreactor scale and 397.57 µM of 3’-PN (135.33 mg/L) and 25.61 µM of 6-PN (8.72 mg/L) were obtained. To the best of our knowledge, this work represents the first report of <em>de novo</em> production of PN compounds using <em>E. coli</em> as a chassis.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"405 ","pages":"Pages 215-228"},"PeriodicalIF":4.1,"publicationDate":"2025-05-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144191777","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":"Substitution of reactive centre loop residues from C1 esterase inhibitor increases the inhibitory specificity of alpha-1 antitrypsin for plasma kallikrein","authors":"Sangavi Sivananthan ,&nbsp;S. Ameer Ahmed ,&nbsp;Ammaar M. Baig ,&nbsp;Varsha Bhakta ,&nbsp;William P. Sheffield","doi":"10.1016/j.jbiotec.2025.05.013","DOIUrl":"10.1016/j.jbiotec.2025.05.013","url":null,"abstract":"<div><div>C1 esterase inhibitor (C1INH) is a member of the serpin superfamily of proteins and controls plasma kallikrein (Pka). Purified C1INH concentrates are effective in controlling C1INH deficiency (hereditary angioedema, HAE). Because C1INH is a relatively slow inhibitor of Pka, we sought to develop a more effective inhibitor by exchanging reactive centre loop (RCL) residues in another serpin, alpha-1 antitrypsin (AAT) variant M358R, with the corresponding residues of C1INH. Novel, soluble, N-terminally hexahistidine-tagged variants were expressed in <em>E. coli</em>, purified by nickel chelate chromatography, and characterized kinetically. <strong>A</strong>AT/<strong>C</strong>1INH loop exchange mutants were designated by the RCL residues exchanged using the reactive centre P1-P1′ convention. Maximal exchange mutant AC (10–4′) inhibited Pka 78-fold and activated Factor XI (FXIa) 350-fold less rapidly than AAT M358R. Eleven additional variants were expressed, restoring AAT residues stepwise. The most selective variant was AC (10–3/4′), which restored AAT residues from P2-P3′ compared to AC (10–4′), and inhibited Pka 1.9-fold more rapidly, and FXIa 1.6-fold less rapidly, for a gain in selectivity of 2.8-fold (p &lt; 0.0001), without increasing the stoichiometry of inhibition (SI). The most active variant was AC (10−3), in which both the rate of Pka and FXIa inhibition were elevated relative to AAT M358R values, without SI elevation. Other variants exhibited slower reaction rates and/or elevated SI values. These results indicate that RCL exchanges can be productively employed to change serpin specificity and selectivity, but that the most effective exchanges may not be contiguous due to cooperativity between RCL residues.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"405 ","pages":"Pages 205-214"},"PeriodicalIF":4.1,"publicationDate":"2025-05-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144170410","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":"pH-driven metabolic reprogramming in Bacillus velezensis 83 regulates metabolite synthesis and sporulation: A transcriptional approach for bioprocess development","authors":"Lorena Yamileth Balón-Rosas,&nbsp;Agustín Luna-Bulbarela,&nbsp;Leobardo Serrano-Carreón,&nbsp;Enrique Galindo","doi":"10.1016/j.jbiotec.2025.05.016","DOIUrl":"10.1016/j.jbiotec.2025.05.016","url":null,"abstract":"<div><div><em>Bacillus velezensis</em> 83 (<em>Bv</em>83) spores are the active ingredient in Fungifree AB, an agricultural biocontrol agent whose industrial production is negatively affected by poly-γ-glutamic acid (γ-PGA) biopolymer synthesis at pH 6.8. At pH 5, γ-PGA production and sporulation are suppressed, but the latter is restored through pH-shift to 6.8 after glucose depletion. This work presents a comprehensive <em>Bv</em>83 spore bioprocess development through physiological and transcriptional analyses. At pH 5, <em>Bv</em>83 present a strategic response to pH stress by reducing its growth rate and redirecting energy towards survival rather than differentiation (downregulating lipopeptide and γ-PGA production genes) and maintaining SigB-mediated stress response rather than sporulation. Transcriptome analysis revealed downregulation of <em>comX</em> and <em>phrC</em> genes at pH 5, indicating sporulation limitation from insufficient signaling molecule production. Indeed, our results confirmed the essential role of the competence and sporulation factor (CSF) in differentiation of elongated cells into forespores. Furthermore, spent medium addition from high-cell-density cultures induced complete sporulation at pH 5, suggesting critical metabolite concentrations (besides CSF) are required. A pH-shift strategy during fed-batch cultivation suppressed γ-PGA synthesis, leading to enhanced mixing and oxygen transfer. Moreover, this strategy led to a 2.6-fold increase in spore productivity (7.86 ×10¹⁰ spores L⁻¹ h⁻¹) compared to a batch at pH 6.8, reducing operational costs. This research has identified pH-regulated metabolic networks, establishing a foundation for designing efficient, cost-effective industrial-scale <em>B. velezensis</em> fermentation strategies that comply with regulatory requirements for biocontrol applications.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"405 ","pages":"Pages 191-204"},"PeriodicalIF":4.1,"publicationDate":"2025-05-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144170592","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}