Enzyme and Microbial Technology最新文献

{"title":"Breaking the CREB–CBP alliance: Progress, challenges, and therapeutic promise of small-molecule and peptide disruptors of the pKID–KIX interaction","authors":"Hassan A. Rudayni","doi":"10.1016/j.enzmictec.2025.110734","DOIUrl":"10.1016/j.enzmictec.2025.110734","url":null,"abstract":"<div><div>The kinase-inducible domain (pKID) of the transcription factor CREB engages the KIX domain of the co-activator CBP/p300 to drive signal-dependent gene expression that underpins cell proliferation, metabolism and survival. Aberrant CREB–CBP signaling is now implicated in numerous cancers and metabolic disorders, yet until recently the shallow, dynamic pKID–KIX interface was viewed as chemically intractable. This review integrates two decades of progress that overturns that paradigm. We first dissect the structural and allosteric features of KIX that enable coupled folding-and-binding of pKID and reveal hidden ligandable pockets. We then survey the expanding inhibitor repertoire—from early micromolar naphthols (KG-501) through nanomolar naphthamides (666−15) and orally tractable pro-drugs, to high-affinity stapled and D-peptide mimetics—highlighting the assays, structure–activity relationships and pharmacokinetic optimization that have driven each advance. Biophysical and computational insights, including ¹⁹F NMR ligandability maps and millisecond-scale molecular-dynamics trajectories, are shown to guide next-generation design and machine-learning pipelines. Pre-clinical data demonstrate that disrupting CREB–CBP selectively suppresses tumor growth with favorable tolerability, and we outline opportunities for combination therapies, degrader strategies and indication expansion into metabolic and neurocognitive disease. Collectively, these findings position the CREB–CBP interaction as a tractable, multi-modal drug target poised for first-in-human exploration.</div></div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"191 ","pages":"Article 110734"},"PeriodicalIF":3.7,"publicationDate":"2025-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144841939","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Exploring the role of catalytic triad Ser-His-Asp on the dynamics of the Lactobacillus helveticus cell-envelope proteinases","authors":"Li Chen ,&nbsp;Ni Lei ,&nbsp;Guanli Du ,&nbsp;He Chen ,&nbsp;Guowei Shu","doi":"10.1016/j.enzmictec.2025.110723","DOIUrl":"10.1016/j.enzmictec.2025.110723","url":null,"abstract":"<div><div>Lactic acid bacterium (LAB) hydrolyzes milk proteins into small bioactive peptides to flourish milk nutrition value. Cell-envelope proteinases (CEPs) are vitally important to bacterial growth, the texture and flavor formation, and the generation of bioactive peptides in fermented milk. Previous literature suggested PR domain of CEP was responsible for the catalytic activity. This study aims to explore the CEP molecular mechanism by delineating the catalytic triad Ser-His-Asp active sites of PR domain in <em>Lactobacillus helveticus</em> CNRZ32 with the aid of homology modeling, molecular docking and dynamics analysis. These results proved that catalytic triads were involved in the PR activation and the catalytic residues Ser608 and His270 appeared to be the core of catalytic process. Our study gained novel insights on the catalytic mechanism of CEP of <em>L. helveticus</em> CNRZ32 which would be a pioneer to facilitate the development of dairy product industry.</div></div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"191 ","pages":"Article 110723"},"PeriodicalIF":3.4,"publicationDate":"2025-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144679800","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A new cold-active transglutaminase: Discovery, computational insights, and recombinant expression","authors":"Han Liu ,&nbsp;Saisai Ding ,&nbsp;Corinne Nardin ,&nbsp;Yi Zhang","doi":"10.1016/j.enzmictec.2025.110712","DOIUrl":"10.1016/j.enzmictec.2025.110712","url":null,"abstract":"<div><div>Transglutaminases (TGases) are versatile enzymes widely applied in food, biomedical, and material sciences, but the cold-active TGases are underexplored despite their significance in low-temperature bioprocessing. This study reports the identification, computational characterization, and recombinant expression of a new transglutaminase (akTGase) derived from the Antarctic krill transcriptome. Bioinformatics analysis revealed that akTGase was estimated of 84.76 kDa, possesses structural traits consistent with cold adaptation, including elevated hydrophilicity, high levels of methionine and aspartic acid but low arginine content, and high proportions of flexible regions. Molecular dynamics simulations at 4 °C and 25 °C showed enhanced surface flexibility and conformational stability at low temperature. Recombinant akTGase was produced in <em>E</em>. <em>coli</em>, recovered from inclusion bodies via stepwise dialysis, and assayed for enzymatic activity. The refolded akTGase displayed significantly higher specific activity at 4 °C than at 25 °C, confirming its cold-active nature. This work highlights a successful strategy combining transcriptomic mining, structural prediction, and experimental validation for the discovery of psychrophilic enzymes, particularly TGase, offering promising potential for sustainable applications in cold-adapted biocatalysis.</div></div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"191 ","pages":"Article 110712"},"PeriodicalIF":3.4,"publicationDate":"2025-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144687466","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Modulating dioxygenase and hydroperoxide isomerase activities in Burkholderia thailandensis lipoxygenase.","authors":"Ruth Chrisnasari, Roelant Hilgers, Guanna Li, Jean-Paul Vincken, Willem J H van Berkel, Marie Hennebelle, Tom A Ewing","doi":"10.1016/j.enzmictec.2025.110709","DOIUrl":"10.1016/j.enzmictec.2025.110709","url":null,"abstract":"<p><p>Lipoxygenases (LOXs) are enzymes that catalyze the regioselective dioxygenation of polyunsaturated fatty acids (PUFAs), leading to the formation of fatty acid hydroperoxides (FAHPs). In addition to dioxygenase activity, some eukaryotic LOXs exhibit hydroperoxide isomerase (HPI) activity under specific conditions, resulting in the production of structurally diverse compounds such as epoxy alcohols and ketones. Until now, the presence of HPI activity in bacterial LOXs has not been documented. In this study, we investigated the HPI activity of LOX from Burkholderia thailandensis (Bt-LOX) and examined the effects of reaction conditions on its catalytic profile using three different C18 PUFA substrates. The results demonstrated that Bt-LOX exhibits significant HPI activity, especially at high enzyme concentrations, with ketone formation showing strong substrate dependence. Oxygen level was identified as a critical factor in directing the catalytic performance of Bt-LOX: HPI activity was inhibited under O₂-saturated conditions and enhanced under O₂-limited conditions. These findings establish Bt-LOX as the first bacterial LOX reported to exhibit pronounced HPI activity, and highlights its expanded potential for biocatalytic applications.</p>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"191 ","pages":"110709"},"PeriodicalIF":3.7,"publicationDate":"2025-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144811958","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Isolation, cloning, and characterization of a novel GH5 cellulase from yak rumen metagenome for enhanced lignocellulose hydrolysis in biofuel production and ruminant feed utilization","authors":"Ibrahim Bature ,&nbsp;Zeyi Liang ,&nbsp;Wu Xiaohu ,&nbsp;Feng Yang ,&nbsp;Yayuan Yang ,&nbsp;Pengcheng Dong ,&nbsp;Xuezhi Ding","doi":"10.1016/j.enzmictec.2025.110737","DOIUrl":"10.1016/j.enzmictec.2025.110737","url":null,"abstract":"<div><div>Lignocellulosic biomass is a promising feedstock for biofuel production, but its complex structure, including cellulose and hemicellulose, challenges efficient enzymatic hydrolysis. <em>CelyA</em>, a novel cellulase from the yak rumen, has potential for improving biomass degradation and biofuel production. The <em>CelyA</em> gene was cloned, expressed, and purified. Biochemical characterization included assessments of pH, temperature, and salt tolerance. The enzyme's substrate specificity was tested on crystalline cellulose, CMC, and other polysaccharides. <em>In vitro</em> rumen fermentation was performed to evaluate its effect on fiber digestibility and microbial community composition. Biofuel production was tested by hydrolyzing maize, rice, and wheat straw. <em>CelyA</em> demonstrated optimal activity at pH 6.0 and 40.0 °C and maintained moderate stability across a wide pH range (3.0–12.0), retaining measurable activity even under strongly acidic and alkaline conditions. The enzyme demonstrated excellent salt tolerance, retaining 91.0 % activity in 1.0 M NaCl. <em>CelyA</em> efficiently degraded maize straw in hydrolysis assays, producing 7.2 µmol/L of reducing sugars. <em>In vitro</em> rumen fermentation with <em>CelyA</em> increased fiber digestibility by 8.3 % for maize straw, 14.5 % for rice straw, and 2.7 % for wheat straw. Gas production also increased significantly, with maize straw showing a 91.3 % increase. 16S rRNA sequencing revealed selective enrichment of <em>Ruminococcus</em> and <em>Prevotella</em>, key cellulolytic microbes. <em>CelyA</em> demonstrates strong potential for biofuel production, efficiently hydrolyzing lignocellulosic biomass and enhancing ruminal fiber digestibility. Its stability, salt tolerance, and substrate specificity make it a valuable enzyme for biofuel production and livestock feed optimization.</div></div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"191 ","pages":"Article 110737"},"PeriodicalIF":3.7,"publicationDate":"2025-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144907276","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Dual-acting single-engineered hybrid-architectured promoters enhance and convert expressions into multi-carbon source-regulated systems in Komagataella phaffii","authors":"Beste Avcı ,&nbsp;Pınar Çalık","doi":"10.1016/j.enzmictec.2025.110713","DOIUrl":"10.1016/j.enzmictec.2025.110713","url":null,"abstract":"<div><div>Ethanol, glycerol, methanol, and acetate are sustainable carbon sources (SCSs) used as substrates for biochemical production. Cells are simultaneously exposed not to a single but multiple external stimuli. SCSs as substrates and co-substrates must be directed/redirected into fermentations. We need <em>de novo</em> <strong>e</strong>ngineered <strong>p</strong>romoters inducible with multi-carbon sources. Core to this conceptual advance is the development of novel methodologies for integrating SCSs into fermentations through engineering transcriptional machinery-element interactions with multiple transcriptional switches, each designed with directed transcription factor (TF) binding site (TFBS)-TF interactions in <em>Komagataella phaffii</em> (<em>Pichia pastoris</em>). <strong>D</strong>ual-<strong>a</strong>cting <strong>s</strong>ingle-<strong>e</strong>ngineered <strong>p</strong>romoters (DASEPs) were designed on <em>alcohol dehydrogenase 2</em> (<em>ADH2</em>) hybrid-architectured promoter layout with two directed synthetic TFBS-TF interactions, function as transcriptional switches to drive SCS-induced upregulated- and/or rewired- transcription and expression. Using cross-yeast analogies, we predicted the master TFs <em><strong>(i)</strong></em> Cat8 on ethanol and methanol and <em><strong>(ii)</strong></em> Hap1 and Hap2/3/4/5 complex on the SCSs. Using single-acting single-engineered promoters (SASEPs) carrying synthetic TFBS-Cat8 transcriptional switch constructed on the base promoter <em>ADH2</em> architecture, we generated DASEP₁ and DASEP₂ on the hybrid-architectured SASEP₃ layout with synthetic TFBS-Hap1 and TFBS-Hap2/3/4/5 transcriptional switches, respectively. DASEP₁ and DASEP₂ performances tested by eGFP expression measurements in SCSs, outcompeted SASEPs and compared to SASEP₃, respectively, <em><strong>(i)</strong></em> 8.2- and 6.5-fold on glycerol, <em><strong>(ii)</strong></em> 2.7- and 2.6-fold on 2 % (v/v) ethanol, <em><strong>(iii)</strong></em> 3.9- and 4.0-fold on 1 % (v/v) ethanol, <em><strong>(iv)</strong></em> 3.6- and 4.2-fold on 1 % (v/v) methanol, and <em><strong>(v)</strong></em> 3.7- and 2.8-fold on acetate. In contrast, lower cell concentrations indicated the metabolic burden of eGFP expression on the metabolic engineered <em>K. phaffii</em> cells constructed with DASEPs.</div></div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"191 ","pages":"Article 110713"},"PeriodicalIF":3.7,"publicationDate":"2025-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144721444","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Elucidation of biochemical attributes and enzymatic activity of agarase from Saccharophagus degradans 2–40","authors":"Anoth Maharjan ,&nbsp;Beom Soo Kim","doi":"10.1016/j.enzmictec.2025.110733","DOIUrl":"10.1016/j.enzmictec.2025.110733","url":null,"abstract":"<div><div><em>Saccharophagus degradans</em> 2–40 exhibits agarolytic activity, effectively degrading agar into galactose. Both endo- and exo-agarase, as well as neoagarobiose hydrolase (NABH), play important roles in agar saccharification for the production of monosugars. This study characterizes a novel agarase enzyme from <em>S. degradans</em> 2–40, a marine bacterium renowned for its exceptional polysaccharide-degrading capabilities. We hypothesized that this strain would harbor an efficient and robust agarase with desirable properties for saccharification processes. Following isolation and purification, the agarase underwent biochemical analysis, revealing optimal activity at moderate temperatures and a broad pH range. Furthermore, the fusion of Aga50D with NABH enhanced the catalytic efficiency from 1.873 ± 0.22 (mg/mL)⁻¹s⁻¹ to 4.826 ± 0.19 (mg/mL)⁻¹s⁻¹. In contrast to chemical hydrolysis, enzymatic treatment using agarase offers a more selective, eco-friendly, and high-yield alternative, minimizing by-product formation and preserving functional sugar moieties. The enzyme's ability to produce neoagarobiose (NA2) as its primary product, without any intermediates, makes it an attractive biocatalyst for the production of bioactive oligosaccharides. This study highlights the potential of <em>S. degradans</em> 2–40 as a valuable source of enzymes for industrial biotechnology applications, particularly in the sustainable conversion of marine biomass into high-value products.</div></div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"191 ","pages":"Article 110733"},"PeriodicalIF":3.7,"publicationDate":"2025-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144826545","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Ethyl lactate synthesis in organic media using a magnetic supported CALB","authors":"Paula Nicolás ,&nbsp;Verónica L. Lassalle ,&nbsp;María Luján Ferreira","doi":"10.1016/j.enzmictec.2025.110732","DOIUrl":"10.1016/j.enzmictec.2025.110732","url":null,"abstract":"<div><div>The growing demand for sustainable chemical processes has spurred interest in enzymatic synthesis, particularly for valuable compounds like ethyl lactate. Traditional chemical methods often suffer from drawbacks, highlighting the potential of enzymatic catalysis using immobilized lipases. This study evaluated the performance of magnetic biocatalyst, prepared by immobilizing <em>Candida antarctica</em> lipase B (CALB) on magnetic nanoparticles, for the batch synthesis of ethyl lactate in hexane. Initial experiments using free CALB and commercial Novozym435 proved problematic due to enzyme denaturation and support instability, respectively. While titration-based methods for monitoring the reaction were found to be unreliable due to lactic acid's complex behavior in the reaction medium, titratable acidity reduction suggested an optimal lactic acid to ethanol molar ratio of 1/10. Subsequent HPLC analysis revealed that the magnetic biocatalyst maintained a consistent conversion (%) at higher lactic acid concentrations (up to 17 mg/mL at 45°C, with conversion above 60 % in 5 h), demonstrating its potential for processing larger amounts of substrate. The initial reaction rate was estimated to be 3.8 mM/h. The study also identified experimental challenges in accurate lactic acid quantification and potential catalyst degradation. In conclusion, the magnetic CALB biocatalyst shows promising activity and stability for ethyl lactate synthesis, especially at higher substrate loads, paving the way for further optimization and application in sustainable production.</div></div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"191 ","pages":"Article 110732"},"PeriodicalIF":3.7,"publicationDate":"2025-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144828379","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Characterization of a GH10 family thermophilic, alkali- and salt-tolerant xylanase from Xinjiang salt lake","authors":"Rong-Huan Song ,&nbsp;Dan Zhu ,&nbsp;Zhong-qiao Yang ,&nbsp;Jian-Ling Li ,&nbsp;Zheng-Feng Yang ,&nbsp;Zhi-Hua Lv ,&nbsp;Kai-Qing Xie ,&nbsp;Li-Quan Yang ,&nbsp;A. Zhou-Cun ,&nbsp;Peng Sang ,&nbsp;Yi-Rui Yin","doi":"10.1016/j.enzmictec.2025.110693","DOIUrl":"10.1016/j.enzmictec.2025.110693","url":null,"abstract":"<div><div>Xylanase is extensively employed in the food, feed, and paper sectors, with those derived from extreme environments offering distinct advantages. This study identified a novel xylanase gene (designated <em>xynaes</em>) through metagenomic analysis of samples from Aiting Lake, Xinjiang, China. Cloned and expressed in Escherichia coli after PCR amplification. The recombinant protein was purified using Ni-NTA affinity chromatography XynAES demonstrated optimal activity at pH 8.0 and 65 °C, its half-life (T_1/2) was 120 min. XynAES preserved over 80 % residual activity after 12 h in pH 6.0–9.0 buffer. Its activity was enhanced to 132 % and 135 % in the presence of 1 mM Mg²⁺ and Zn²⁺, respectively. Additionally, XynAES maintained over 60 % relative activity in 0–3.0 M NaCl and its Km and Vmax of XynAES were determined to be 3.23 mg/mL and 72.46 μmol/min/mg, respectively. It is worth noting that the main products of XynAES enzymatic hydrolysis of xylan are xylose disaccharides and xylose tetrasaccharides, and XynAES shows obvious activity against the pre-treated wheat bran. In summary, XynAES is a thermophilic, alkali-tolerant, and salt-resistant xylanase, signifying its potential applications in the feed, food baking, paper manufacturing, and prebiotic production industries.</div></div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"190 ","pages":"Article 110693"},"PeriodicalIF":3.4,"publicationDate":"2025-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144262912","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Functional characterization of CEL3C reveals its critical role in regulating cellulase gene expression in Trichoderma reesei Rut C30","authors":"Lu Wang ,&nbsp;Xiao He ,&nbsp;Tian Tian ,&nbsp;Jian Cheng ,&nbsp;Ruolan Cao ,&nbsp;Jie Hou ,&nbsp;Hui Lin ,&nbsp;Yonghao Li","doi":"10.1016/j.enzmictec.2025.110706","DOIUrl":"10.1016/j.enzmictec.2025.110706","url":null,"abstract":"<div><div>The nuclear-localized β-glucosidase CEL3C in <em>Trichoderma reesei</em> plays a pivotal role in cellulase regulation, though its mechanism remains poorly understood. To address this, we disrupted CEL3C in the hypercellulolytic strain <em>T. reesei</em> Rut C30 via CRISPR-Cas9 and evaluated cellulase production under sophorose-rich MGD induction. Deletion of CEL3C significantly enhanced total cellulase activity by 31.28 % (<em>p</em> &lt; 0.05), with β-glucosidase, endoglucanase, and cellobiohydrolase activities increasing by 94.97 %, 19.40 %, and 28.99 %, respectively. These improvements were driven by transcriptional upregulation of core cellulase genes (CEL7A: 2.01-fold; CEL6A: 1.5-fold; CEL12A: 2.0-fold; CEL5A: 1.32-fold) and β-glucosidases (CEL3A: 6.41-fold; CEL3B: 5.02-fold), confirming transcriptional-level control as the dominant regulatory mechanism. Transcriptomic profiling identified 688 differentially expressed genes (399 upregulated, 299 downregulated), with key changes including activation of transcriptional activators XYR1 (59.6 % increase), ACE3 (75.49 % increase), and RXE1 (161.56 % increase), suppression of repressors RCE1 (65.86 % decrease) and RCE2 (65.23 % decrease), and induction of sugar transporters (TrireC30_133589: 13.41-fold) and ER chaperones (BIP1: 1.26-fold; PDI1: 1.55-fold). These alterations collectively enhanced inducer uptake, enzyme maturation, and secretion while alleviating MAPK-mediated repression (TMK2: 110.54 % decrease). Intracellular sugar profiling revealed that gentiobiose and cellobiose were undetectable in the <em>T. reesei</em> ΔCEL3C, whereas glucose and sophorose levels increased by 31.71 % and 13.45 % (<em>p</em> &lt; 0.05), respectively. These results suggest that CEL3C deletion enhances β-glucosidase-mediated hydrolysis of disaccharides into glucose and possibly promotes sophorose formation via transglycosylation. In parallel, the upregulation of disaccharide transporters may facilitate sophorose uptake. Together, these two mechanisms contributed to the intracellular enrichment of sophorose, thereby amplifying cellulase gene induction and enzyme production. Our findings establish CEL3C as a dual-function nuclear regulator that balances cellulase synthesis through transcriptional and enzymatic pathways, providing actionable targets for engineering <em>T. reesei</em> with optimized industrial cellulase yields.</div></div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"190 ","pages":"Article 110706"},"PeriodicalIF":3.4,"publicationDate":"2025-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144549872","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}