Journal of biotechnology最新文献

{"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":"2026-02-01","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}

{"title":"Methanol-induced metabolic disturbances in protein expression in Pichia pastoris: A review of the interaction between amino acid synthesis and higher alcohol pathways","authors":"Lifen He ,&nbsp;Danling Tang ,&nbsp;Fan Feng ,&nbsp;Tao Hong ,&nbsp;Mingjing Zheng ,&nbsp;Xiping Du ,&nbsp;Yanbing Zhu ,&nbsp;Yuanfan Yang ,&nbsp;Zedong Jiang ,&nbsp;Fan He ,&nbsp;Lijun Li ,&nbsp;Zhipeng Li ,&nbsp;Hui Ni ,&nbsp;Qingbiao Li","doi":"10.1016/j.jbiotec.2025.11.015","DOIUrl":"10.1016/j.jbiotec.2025.11.015","url":null,"abstract":"<div><div><em>Pichia pastoris</em> is a prominent host for recombinant protein production, utilizing the methanol-inducible Alcohol Oxidase 1 (AOX1) promoter to express thousands of heterologous proteins. However, methanol induction, while driving high-level protein expression, simultaneously imposes a pronounced metabolic burden. This review highlights that a balanced and ample supply of amino acids is indispensable for efficient protein synthesis. The induction process leads to the accumulation of by-products, notably higher alcohols, which can negatively impact protein quality. The dynamic interplay between these two pathways is not fully understood. In addition, the lack of quantitative models for their competition over shared precursors hinders rational metabolic engineering and impedes the industrial exploitation of <em>P. pastoris</em>. Therefore, this review systematically dissects the link between methanol induction, amino acid metabolism, and higher alcohol synthesis. It specifically examines how methanol-triggered perturbations in amino acid metabolism propagate to modulate higher alcohol formation. Special attention is given to key metabolic nodes, such as pyruvate and α-ketoisovalerate, where carbon flux diverges between biosynthesis and by-product formation. This review provides significant implications for a deeper understanding of the metabolic processes in <em>P. pastoris</em>, offering comprehensive mechanistic insights and guidance for optimizing yeast fermentation and enhancing recombinant protein production.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"410 ","pages":"Pages 57-66"},"PeriodicalIF":3.9,"publicationDate":"2026-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145622160","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":"Enhancing oxidative stress defense to improve docosahexaenoic acid production in Schizochytrium sp. HX-308","authors":"Chunxiao Yan ,&nbsp;Shuai Zhang ,&nbsp;Zhihao Huang ,&nbsp;Fei Du ,&nbsp;Qian Xu ,&nbsp;Wang Ma ,&nbsp;Guang Yang ,&nbsp;Xiaoman Sun","doi":"10.1016/j.jbiotec.2025.12.005","DOIUrl":"10.1016/j.jbiotec.2025.12.005","url":null,"abstract":"<div><div>As an excellent host for docosahexaenoic acid (DHA) biosynthesis, <em>Schizochytrium</em> sp. still faces inherent limitations in DHA yield due to oxidative damage induced by reactive oxygen species (ROS). To effectively reduce the ROS levels, the optimal combination of mixed antioxidants (52.95 mg L⁻¹ phytic acid, 32.00 mg L⁻¹ licorice extract, 36.28 mg L⁻¹ ascorbic acid palmitate) was selected by single factor experiment combined with response surface optimization. The DHA titer increased from 24.0 g L⁻¹ to 33.5 g L⁻¹. However, exogenous addition of antioxidants results in increased the cost of DHA production. Enhancing the intracellular antioxidant capacity through gene engineering has become another feasible strategy. Transcriptome analysis indicated that the transcriptional levels of antioxidant-related genes <em>SOD1</em> (superoxide dismutase), <em>APX5</em> (ascorbic acid peroxidase), and <em>GST3</em> (glutathione S-transferase) were significantly upregulated during DHA biosynthesis. Then, the engineered strain SOD1-APX5-GST3–2 was constructed by overexpressing the three genes, and the DHA and lipid titers were 33.1 g L⁻¹ and 64.1 g L⁻¹, respectively. Scale-up in a 5 L bioreactor obtained 32.5 g L⁻¹ DHA and 63.5 g L⁻¹ lipid, and the ROS level decreased by 30.5 %, demonstrating engineered strain's stability and laying the foundation for industrial-scale production. This study verified that reducing ROS levels promoted DHA biosynthesis in <em>Schizochytrium</em> sp. from the two dimensions of exogenous antioxidants and gene engineering, which provided novel insights for industrial lipid production.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"410 ","pages":"Pages 162-172"},"PeriodicalIF":3.9,"publicationDate":"2026-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145756688","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":"Heavy ion beam irradiation-induced mutational profiles in Saccharomyces cerevisiae and their dependencies on dose and intracellular state inform an enhanced microbial breeding strategy","authors":"Xiaopeng Guo ,&nbsp;Xiuyue Xiao ,&nbsp;Xuexia Dong ,&nbsp;Runsheng Yin ,&nbsp;Shengli Zhang ,&nbsp;Miaomiao Zhang ,&nbsp;Guanghong Luo ,&nbsp;Yonggang Wang ,&nbsp;Dong Lu","doi":"10.1016/j.jbiotec.2025.12.015","DOIUrl":"10.1016/j.jbiotec.2025.12.015","url":null,"abstract":"<div><div>Efficient microbial mutagenesis using heavy-ion beam (HIB) radiation is crucial for breeding. Here, <em>Saccharomyces cerevisiae</em> was irradiated with HIB across medium and high doses. Based on 65 randomly selected isolates, we systematically characterized the mutagenic features and preliminarily explored the influence of gene transcriptional activity on mutation susceptibility, while also modulating the intracellular state to optimize strain breeding. High-dose irradiation (120–210 Gy) resulted in a mutation frequency more than double that of the medium-dose (90 Gy), with minimal overlapping mutations between doses. Although mutation site numbers correlated with chromosome length broadly, they were not uniformly distributed at a finer scale. The overall expression of genes associated with mutation sites moderately exceeded the genome-wide background level (<em>p</em> &lt; 0.05). By coupling radiation with osmotic stress, osmoregulatory-related genes were induced to express highly during irradiation. The proportion of osmotolerant mutants obtained from each coupled treatment group (averaging 27.62 %) was higher than that from the radiation-only group (11.43 %). Inference and validation indicated that early selection pressure alone could not fully account for this improvement, highlighting the importance of the intracellular state. Compared to radiation alone, coupled radiation-osmotic stress increased the distribution of mutations in osmotically inducible osmoregulatory-related genes. We propose that the enhanced transcriptional activity may alter local chromatin conformation, together with pre-activation of shared osmotic-radiation response genes, reshape the damage-repair-mutagenesis balance. The coupled treatment produced genetically stable, highly osmotolerant mutants with mutations synergistically regulating carbon metabolism, ion homeostasis, cell adhesion, and DNA replication. This work supports developing high-efficiency microbial breeding strategies.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"410 ","pages":"Pages 236-257"},"PeriodicalIF":3.9,"publicationDate":"2026-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145804626","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":"Enhancing heterotrophic lutein production in Chlorella protothecoides through combined phytohormone and nitrogen strategies","authors":"Jiaxin Li ,&nbsp;Weiyang Zhao ,&nbsp;Yunlei Fu ,&nbsp;Ranxuan Li ,&nbsp;Jin Liu ,&nbsp;Shufang Yang ,&nbsp;Jie Zheng ,&nbsp;Haijin Mou ,&nbsp;Han Sun","doi":"10.1016/j.jbiotec.2025.12.001","DOIUrl":"10.1016/j.jbiotec.2025.12.001","url":null,"abstract":"<div><div>Large-scale production of microalgal lutein faces challenges due to limited biomass and lutein content. In this work, we developed an exponential fed-batch model to achieve heterotrophic high-density growth of <em>Chlorella protothecoides</em> CS-41. Then, indole-3-acetic acid (IAA) and high nitrogen were combined to promote lutein accumulation <em>in situ</em>. Fed-batch culture enhanced the TCA cycle of <em>C. protothecoides</em> CS-41 to benefit cell growth while increasing carotenoid flux to facilitate lutein accumulation through redirecting carbon flux towards lutein biosynthesis, improved by IAA. Also, high nitrogen increased lutein content through modulation of <em>CMS</em> (carotenoid methyltransferase synthase), <em>CRTISO</em> (carotenoid isomerase), <em>LCYB</em> (lycopene β-cyclase), <em>ZEP</em> (zeaxanthin epoxidase), and <em>VDE</em> (violaxanthin de-epoxidase). Finally, under heterotrophic conditions, combining 200 mM IAA with 3.6 g/L urea improved pyruvate and 3-phosphoglyceric acid and then boosted lutein content and productivity to 3.27 mg/g and 20.38 mg/L/d, respectively, by increases of 58 % and 56.7 %. Addressing issues including density dilution, container transfer, and <em>in-situ</em> lutein accumulation under heterotrophic high-density conditions offers a cost-effective solution.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"410 ","pages":"Pages 125-136"},"PeriodicalIF":3.9,"publicationDate":"2026-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145681983","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":"Aluminum phthalocyanine chloride loaded poly (lactic-co-glycolic acid) nanoparticles based photodynamic therapy for antibacterial and anticancer applications","authors":"Anum Jamil ,&nbsp;Aleena Zahid ,&nbsp;Rafaqat Ali Khan ,&nbsp;Muhammad Saleem ,&nbsp;Syed Mujtaba Ul Hassan ,&nbsp;Shahzad Anwar ,&nbsp;Qura Tul Ain","doi":"10.1016/j.jbiotec.2025.12.014","DOIUrl":"10.1016/j.jbiotec.2025.12.014","url":null,"abstract":"<div><div>Photodynamic therapy (PDT) and antibacterial Photodynamic therapy (aPDT) have become a viable non-invasive treatment against cancer and bacterial infections. By using light-activated photosensitizers (PS), PDT effectively destroys cancer cells and pathogenic bacteria. However, many PS agents, such as Aluminum Phthalocyanine chloride (AlPc), face limitations like poor solubility and low bioavailability. To overcome these challenges, AlPc-loaded PLGA nanoparticles (AlPc NPs) have been synthesized, where PS is encapsulated in Poly (lactic-co-glycolic acid) (PLGA) to enhance drug delivery and therapeutic efficacy. PLGA was selected for its well-established biomedical applications, including targeted drug delivery and antibacterial activity. This study focuses on synthesizing 200 nm AlPc NPs by using the single emulsion solvent evaporation method and characterizing them via spectroscopic and microscopic techniques. This study demonstrates that AlPc NPs' encapsulation efficiency is 82 % and drug loading efficiency is 10 %. The drug release profile shows 52 % release of AlPc NPs within 24 h. Cellular uptake of free AlPc and AlPc NPs was examined using confocal fluorescence imaging. The in vitro investigation used Hep-2C human laryngeal cancer cells to assess the dark and phototoxic effects of free AlPc and AlPc NPs for PDT. MTT results show that the cells treated with AlPc NPs had lower cell viability than cells treated with free AlPc at the same drug doses, which confirms improved efficacy of AlPc NPs. Moreover, AlPc NPs improved the antibacterial potential against the <em>Escherichia coli (E. coli)</em> bacterial strain and increased their bioavailability.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"410 ","pages":"Pages 309-320"},"PeriodicalIF":3.9,"publicationDate":"2026-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145800443","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":"Rational design of a bifunctional glycosyltransferase for enhanced substrate promiscuity and thermostability","authors":"Dekai Liu ,&nbsp;Ruifang Zhang ,&nbsp;Peiqin Chen ,&nbsp;Chunying Jin ,&nbsp;Lingzhi Zhang ,&nbsp;Xiao Gu ,&nbsp;Li Wen ,&nbsp;Lijuan Zhang ,&nbsp;Guangya Zhang ,&nbsp;Zhongkun Wu ,&nbsp;Wei Jiang","doi":"10.1016/j.jbiotec.2025.12.009","DOIUrl":"10.1016/j.jbiotec.2025.12.009","url":null,"abstract":"<div><div>Glycosylation of natural products significantly enhances their physicochemical properties, with glycosyltransferases (GTs) serving as the catalytic core of this biotransformation. Rational design of tailored GTs aligned with green chemistry principles is pivotal for the targeted synthesis of high-value glycosides, yet precise functional engineering remains challenging. This study employed a multi-scale computational strategy (molecular docking, multiple sequence alignment, molecular dynamics simulations) to systematically re-engineer the substrate recognition and stability modules of the bacterial GT BsGT-1. By reverse-engineering the active pocket of a plant-derived hyperpromiscuous GT (FiGT-2), we orthogonally mapped and identified six functional hotspots in BsGT-1. Site-directed mutagenesis and screening yielded the double variant S128T/T229S, achieving dual optimization: improved substrate scope (121–140 % increase in conversion with UDP-Gal/UDP-Rha compared to wild-type) and enhanced thermostability (&gt;70 % residual activity after 4 h at 50℃). Structural dynamics analyses revealed that mutation-induced global conformational rigidity and localized hydrogen-bond network optimization primarily drove thermostability improvement and substrate affinity enhancement, respectively. This work establishes a closed-loop engineering paradigm of \"computational prediction → rational mutagenesis → mechanistic decoding\", providing a scalable framework for precision engineering of GTs and glycoside biomanufacturing.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"410 ","pages":"Pages 285-297"},"PeriodicalIF":3.9,"publicationDate":"2026-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145804717","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":"Lyophilized chloroplasts as molecular capsules: A scalable platform for stable and functional human α-lactalbumin production","authors":"Maryam Ehsasatvatan ,&nbsp;Bahram Baghban Kohnehrouz","doi":"10.1016/j.jbiotec.2025.12.016","DOIUrl":"10.1016/j.jbiotec.2025.12.016","url":null,"abstract":"<div><div>The growing world population and the increasing demand for proteins present significant challenges to food security. Advances in synthetic biology have enabled the development of transplastomic plants engineered to produce milk proteins, offering a promising solution for the large-scale, economical production of alternative protein sources. This study demonstrated the extended stability of human α-lactalbumin (hLA) in freeze-dried transplastomic tobacco chloroplasts. hLA was expressed at high levels, constituting 39.1 % of the total soluble protein in chloroplasts (66.5 mg/g isolated chloroplasts). Western blot analysis of lyophilized tobacco chloroplasts stored at room temperature for up to six months indicated that the hLA protein remained stable, soluble, and properly folded over extended periods. The freeze-drying process of isolated chloroplasts resulted in a 34-fold increase in hLA protein concentrations. Importantly, the biological functionality of purified hLA from lyophilized chloroplasts was confirmed through an <em>in vitro</em> lactose synthesis assay, where recombinant hLA successfully activated galactosyltransferase, leading to efficient lactose production at level comparable to commercial standard. Lyophilized chloroplasts, free from nicotine and bacterial contamination, are suitable candidates for the oral administration of the dietary protein alpha-lactalbumin. These findings suggest that recombinant protein production in transplastomic plants could serve as a viable food source and alternative to conventional sources. Lyophilization further reduces expenses and streamlines downstream processing, purification, and storage. These methods facilitate relevant formulation practices for these compounds to meet the demand-oriented requirements. Future research should prioritize translation of this technology to edible plant hosts to enable direct human consumption and undertake comprehensive <em>in vivo</em> and clinical studies to validate safety, bioavailability, and efficacy.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"410 ","pages":"Pages 266-275"},"PeriodicalIF":3.9,"publicationDate":"2026-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145834083","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":"2026-02-01","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":"Next level of p-phenylene diisothiocyanate-based covalent immobilization of β-D-galactosidase: Technical optimization as an application","authors":"Tabea L. Boehme ,&nbsp;Bernadette Straub ,&nbsp;Anja Oberle ,&nbsp;Ursula Eschenhagen ,&nbsp;Magnus S. Schmidt","doi":"10.1016/j.jbiotec.2025.12.004","DOIUrl":"10.1016/j.jbiotec.2025.12.004","url":null,"abstract":"<div><div>In this study, a continuous lactose hydrolysis process in a fixed-bed reactor was developed using β-galactosidase covalently immobilized on resin beads via 1,4-phenylenediisothiocyanate (PDC) as linker. Process conditions, including temperature, enzyme loading, perfusion speed, and repeated perfusion of the same substrate solution were systematically varied. The highest glucose yields were obtained at 55 °C, with increased yields observed at low perfusion speeds, high enzyme loadings, and upon repeated perfusions. Under optimized cycle perfusion over 72 h, final lactose conversion reached approximately 90 % at 37 °C and 80 % at 22 °C. A hydrolysis process in a fixed-bed reactor was successfully established, although further optimization is required.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"410 ","pages":"Pages 153-161"},"PeriodicalIF":3.9,"publicationDate":"2026-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145742781","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}