Microbial Biotechnology最新文献

{"title":"Genetic Toolbox Expansion Enables Constitutively Fluorescent Lacticaseibacillus rhamnosus for Functional Microbiome Research","authors":"Ilke Van Tente,&nbsp;Marc Blanch-Asensio,&nbsp;Tom Eilers,&nbsp;Dieter Vandenheuvel,&nbsp;Sarah Lebeer,&nbsp;Shrikrishnan Sankaran,&nbsp;Irina Spacova","doi":"10.1111/1751-7915.70405","DOIUrl":"10.1111/1751-7915.70405","url":null,"abstract":"<p><i>Lacticaseibacillus rhamnosus</i> strains are widely recognized for their probiotic potential and relevance in urogenital and gut health. However, their genetic tractability and genetic tools remain limited, hindering functional microbiome research and synthetic biology applications. In this study, we expanded the genetic toolbox for the widely used probiotic strains, <i>L. rhamnosus</i> GR-1 and <i>L. rhamnosus</i> GG, by implementing direct plasmid cloning and testing of a set of genetic elements earlier validated in <i>Lactiplantibacillus plantarum</i>. Among five constitutive promoters (P_tlpA, P_tec, P_cpg, P₄₈ and P₂₃), P_tlpA showed strong promoter activity in <i>L. rhamnosus</i> GR-1. We further characterized this promoter's functionality by incorporating a repressor and assessing its native thermo-responsiveness and stability over time, enhancing its potential for industrial applications. Using these tools, we engineered <i>L. rhamnosus</i> GR-1 with constitutive fluorescence of mCherry, mScarlet3 and sfGFP. The functionality of these fluorescent <i>L. rhamnosus</i> GR-1 strains was shown in a proof-of-concept growth competition experiment with a fluorescent pathogenic <i>Staphylococcus aureus</i> strain. These constitutively fluorescent <i>L. rhamnosus</i> strains, along with the expanded genetic toolkit, offer valuable resources for studying functional properties, such as adhesion, microbe–microbe and host–microbe interactions, and advancing <i>Lactobacillaceae</i> as a chassis for synthetic biology.</p>","PeriodicalId":209,"journal":{"name":"Microbial Biotechnology","volume":"19 7","pages":""},"PeriodicalIF":6.7,"publicationDate":"2026-07-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/1751-7915.70405","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148417113","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Harnessing the PpHog1 Regulatory Network to Engineer a UPR-Adaptive Komagataella phaffii Chassis for High-Copy Secretory Protein Production","authors":"Jiacheng Shi,&nbsp;Zhanqing Lv,&nbsp;Chaoyu Lu,&nbsp;Bohao Fang,&nbsp;Zerui Zou,&nbsp;Mian Zhou","doi":"10.1111/1751-7915.70410","DOIUrl":"10.1111/1751-7915.70410","url":null,"abstract":"<p><i>Komagataella phaffii</i> is one of the most widely used eukaryotic protein expression systems. Increasing the copy number of exogenous genes is a common method to enhance expression efficiency. However, at extremely high copy numbers, the resulting endoplasmic reticulum stress (ERS) can hinder further improvement in the expression efficiency of secreted proteins. And elucidating the underlying molecular mechanisms serves as the foundation for engineering and constructing stress-adapted strains. High-copy secretory expression was first confirmed to adversely affect yeast growth fitness, a phenotype consistent with DTT-induced unfolded protein response (UPR). Screening of a kinase knockout library established that UPR signalling in <i>K. phaffii</i> was modulated by the PpHog1-mediated MAPK pathway. PpHog1, the <i>K. phaffii</i> ortholog of the well-characterized <i>Saccharomyces cerevisiae</i> Hog1 (67% sequence identity), is traditionally known for its roles in osmotic stress and cell wall integrity. Notably, while ScHog1 has also been implicated in ERS regulation, the downstream pathway remains obscure. Further comparison of the PpHog1 interactome with versus without DTT treatment, combined with follow-up genetic knockout/knockdown screens, leads to the identification of two critical UPR regulators: PpLRR-0498 and PpPINT-0120. Subsequently, an engineered UPR-adaptive chassis (UPR-Ad⁺) was constructed by over-expressing these three key regulators. Using human serum albumin (HSA), human calcium-regulated actin binding protein LCP1, and α-amylase as reporters, UPR-Ad⁺ enhanced the production of high-copy secretory proteins by 30%–46%. Collectively, this work advances the understanding of UPR regulation in <i>K. phaffii</i> and provides a strategic basis for designing high-yield strains.</p>","PeriodicalId":209,"journal":{"name":"Microbial Biotechnology","volume":"19 7","pages":""},"PeriodicalIF":6.7,"publicationDate":"2026-07-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/1751-7915.70410","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148417150","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"WC1/WC2–Sre1 Transcriptional Cascade Controls Predation and Chlamydospore Formation in the Nematode-Trapping Fungus Arthrobotrys flagrans","authors":"Yu Zhang,&nbsp;Jiafang Zuo,&nbsp;Qianfei Shi,&nbsp;Peiji Zhao,&nbsp;Hanbo Zhang,&nbsp;Minghe Mo,&nbsp;Guohong Li","doi":"10.1111/1751-7915.70412","DOIUrl":"10.1111/1751-7915.70412","url":null,"abstract":"<p><i>Arthrobotrys flagrans</i> is a nematode-trapping fungus that kills plant-parasitic nematodes (PPNs) using adhesive traps and withstands adverse conditions by producing chlamydospores. It holds promise as a biocontrol agent against PPNs, yet the mechanisms underlying its pathogenicity and chlamydospore formation remain poorly characterized, hampering the development of efficient biocontrol strains. Here, we systematically characterized all eight GATA-type transcription factors in <i>A. flagrans</i> and found that all were significantly upregulated during both trap and chlamydospore formation. These eight regulators exhibit specialized and coordinated functions in hyphal growth, stress responses, pathogenicity, and chlamydospore formation. WC2, WC1, Sre1, AreA, ASD4 and GATA1 positively regulate pathogenicity, whereas Ams2 and NsdD act as negative regulators, likely through the repression of virulence factors and adhesins. Moreover, all eight factors govern chlamydospore number, with WC1, WC2 and Sre1 additionally controlling chlamydospore diameter. Further investigation of their interactions and regulatory relationships revealed a novel, light-independent WC1/WC2–Sre1 transcriptional cascade as a key node within the network governing pathogenicity and chlamydospore formation. Overexpression of <i>WC2</i> significantly increases pathogenicity and chlamydospore production, providing functional validation that WC2 is a precise genetic target for engineering superior biocontrol strains. Overall, this work elucidates the functional atlas of GATA-type transcription factors in <i>A. flagrans</i>, reveals a novel light-signalling co-option mechanism for developmental regulation, and provides both actionable molecular targets for strain improvement and a theoretical framework for advancing the biocontrol of PPNs.</p>","PeriodicalId":209,"journal":{"name":"Microbial Biotechnology","volume":"19 7","pages":""},"PeriodicalIF":6.7,"publicationDate":"2026-07-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/1751-7915.70412","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148417133","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Mind the Porins: Differential Effects of Porin Knockouts and Overexpression on Glucose and Xylose Uptake and Utilization in Pseudomonas putida","authors":"Barbora Popelářová,&nbsp;Nicolas T. Wirth,&nbsp;Daniel C. Volke,&nbsp;Tibor Botka,&nbsp;Pablo I. Nikel,&nbsp;Pavel Dvořák","doi":"10.1111/1751-7915.70406","DOIUrl":"10.1111/1751-7915.70406","url":null,"abstract":"<p>Porins serve as the primary transport channels for substrate molecules across the outer membrane of Gram-negative bacteria. Despite their potential to influence substrate uptake in microbial cell factories, porins are often overlooked in metabolic engineering approaches. In this study, we investigate the impact of modulation of sugar porin expression using laboratory and industrial workhorse <i>Pseudomonas putida</i>. We first examined the <i>P. putida</i> porin repertoire through bioinformatic analysis. Among the two selected porin sets, only the one comprising OprB-I, OprB-II and OprB-III was found to be relevant for glucose catabolism in two biotechnologically important <i>P. putida</i> strains. Functional studies involving gene knockouts, complementation and overexpression revealed that the substrate specificity of <i>P. putida</i> OprB porins extends beyond glucose and includes the non-native substrate xylose. Overexpression of <i>oprB-I</i> alone was sufficient to restore sugar utilization in strains with all three <i>oprB</i> genes knocked out. Notably, when Gcd was active in <i>P. putida</i>, <i>oprB</i>-I overexpression accelerated the utilization of glucose and xylose in mixed sugar conditions through altered sugar uptake and oxidation dynamics. This work exposes the relevance of porins in shaping the uptake of major lignocellulosic sugars and highlights the importance of incorporating outer membrane transport considerations into metabolic engineering strategies for Gram-negative bacteria.</p>","PeriodicalId":209,"journal":{"name":"Microbial Biotechnology","volume":"19 7","pages":""},"PeriodicalIF":6.7,"publicationDate":"2026-07-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/1751-7915.70406","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148381364","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Coumarins Disrupt Cell–Cell Communication and Virulence in Priority Pathogens: Targeting the PQS Signalling System in Pseudomonas aeruginosa","authors":"Dylan Boon,&nbsp;Benjamin O'Rourke,&nbsp;Muireann Carmody,&nbsp;David F. Woods,&nbsp;Antje Gloe,&nbsp;Daniel Platero-Rochart,&nbsp;Pedro A. Sánchez-Murcia,&nbsp;Gerard P. McGlacken,&nbsp;F. Jerry Reen","doi":"10.1111/1751-7915.70404","DOIUrl":"https://doi.org/10.1111/1751-7915.70404","url":null,"abstract":"<p>Cell-to-cell communication in microbial systems is known for its vital role in cellular signalling and gene expression. A specific form termed Quorum Sensing (QS) has received considerable attention since its discovery in the marine symbiont <i>Aliivibrio fischeri</i>. QS-controlled microbial functions are associated with bacterial virulence, pathogenicity, host–microbe interactions, and biofilm development. Interference in these signalling systems can modulate microbial virulence and pathogenicity, and microbial infection caused by drug-resistant pathogens. Plant-derived phytochemicals are considered a promising candidate, with coumarins emerging as significant plant-derived signalling molecules shaping microbiome dynamics and pathogen behaviours from a broad spectrum of ecosystems. Here we explored the role of natural and synthetic coumarin compounds in the control of signalling and virulence traits in <i>Pseudomonas aeruginosa</i> and other priority bacterial pathogens, including the fungal opportunist <i>Aspergillus fumigatus</i>. We uncovered an important ‘hydroxylation-bias’ favouring coumarin, umbelliferone (7-OH), and 6-hydroxy-coumarin (6-OH) in the specific competitive inhibition of the Pseudomonas Quinolone Signal (PQS), associated with reduced activity of a PqsR translational fusion and suppression of pyocyanin production. Conversely, while esculetin (6,7-OH) was most effective at Acyl Homoserine Lactone (AHL) QS biosensor inhibition, it did not affect PQS production. Anti-biofilm activity of coumarins against <i>P. aeruginosa</i> was independent of initial attachment but linked to changes in exopolysaccharide production. As the very real threat posed by antimicrobial resistance persists, these data support a role for phytochemicals such as coumarins in delivering an ecological solution to dysbiosis in the host–microbe interaction.</p>","PeriodicalId":209,"journal":{"name":"Microbial Biotechnology","volume":"19 7","pages":""},"PeriodicalIF":6.7,"publicationDate":"2026-06-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/1751-7915.70404","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148322466","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Coumarins Disrupt Cell–Cell Communication and Virulence in Priority Pathogens: Targeting the PQS Signalling System in Pseudomonas aeruginosa","authors":"Dylan Boon,&nbsp;Benjamin O'Rourke,&nbsp;Muireann Carmody,&nbsp;David F. Woods,&nbsp;Antje Gloe,&nbsp;Daniel Platero-Rochart,&nbsp;Pedro A. Sánchez-Murcia,&nbsp;Gerard P. McGlacken,&nbsp;F. Jerry Reen","doi":"10.1111/1751-7915.70404","DOIUrl":"https://doi.org/10.1111/1751-7915.70404","url":null,"abstract":"<p>Cell-to-cell communication in microbial systems is known for its vital role in cellular signalling and gene expression. A specific form termed Quorum Sensing (QS) has received considerable attention since its discovery in the marine symbiont <i>Aliivibrio fischeri</i>. QS-controlled microbial functions are associated with bacterial virulence, pathogenicity, host–microbe interactions, and biofilm development. Interference in these signalling systems can modulate microbial virulence and pathogenicity, and microbial infection caused by drug-resistant pathogens. Plant-derived phytochemicals are considered a promising candidate, with coumarins emerging as significant plant-derived signalling molecules shaping microbiome dynamics and pathogen behaviours from a broad spectrum of ecosystems. Here we explored the role of natural and synthetic coumarin compounds in the control of signalling and virulence traits in <i>Pseudomonas aeruginosa</i> and other priority bacterial pathogens, including the fungal opportunist <i>Aspergillus fumigatus</i>. We uncovered an important ‘hydroxylation-bias’ favouring coumarin, umbelliferone (7-OH), and 6-hydroxy-coumarin (6-OH) in the specific competitive inhibition of the Pseudomonas Quinolone Signal (PQS), associated with reduced activity of a PqsR translational fusion and suppression of pyocyanin production. Conversely, while esculetin (6,7-OH) was most effective at Acyl Homoserine Lactone (AHL) QS biosensor inhibition, it did not affect PQS production. Anti-biofilm activity of coumarins against <i>P. aeruginosa</i> was independent of initial attachment but linked to changes in exopolysaccharide production. As the very real threat posed by antimicrobial resistance persists, these data support a role for phytochemicals such as coumarins in delivering an ecological solution to dysbiosis in the host–microbe interaction.</p>","PeriodicalId":209,"journal":{"name":"Microbial Biotechnology","volume":"19 7","pages":""},"PeriodicalIF":6.7,"publicationDate":"2026-06-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/1751-7915.70404","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148322374","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Engineering Pseudomonas protegens as a Targeted Antifungal-Effector Delivery Chassis via Heterologous Type III Secretion System Reconstitution","authors":"Yuyuan Peng,&nbsp;Simei He,&nbsp;Guofan Zhang,&nbsp;Jiatao Yang,&nbsp;Rong Fan,&nbsp;Chen Li,&nbsp;Jiajie Xie,&nbsp;Xinran Long,&nbsp;Linger Qian,&nbsp;Jiayi Wang,&nbsp;Yanmin He,&nbsp;Hui Cai,&nbsp;Yushan Zhang,&nbsp;Qiuqian Zhang,&nbsp;Shaopeng Pan,&nbsp;Shengbiao Hu","doi":"10.1111/1751-7915.70403","DOIUrl":"10.1111/1751-7915.70403","url":null,"abstract":"<p>This study addresses the core challenge of Fusarium wilt control in agricultural production. We successfully reconstituted a functional heterologous type III secretion system (T3SS) from <i>Photorhabdus luminescens</i> in the biocontrol bacterium <i>Pseudomonas protegens</i> Pf-5, creating an engineered molecular syringe for targeted delivery of antifungal effectors. The system is activated under low-calcium conditions, achieved by cultivation in calcium-limited medium followed by EGTA-mediated chelation of residual Ca²⁺, enabling conditional secretion of effector proteins. By fusing the antifungal protein Bg9562 to the N-terminal secretion signal of the T3SS effector LopT and co-expressing it with the cognate chaperone SlcT, we obtained fluorescence-based evidence for T3SS-dependent delivery of Bg9562 into the hyphae of multiple Fusarium species. The engineered strain exhibited enhanced rhizosphere colonization, promoted plant growth and conferred improved protection against tomato Fusarium wilt, restoring plant height to levels approaching healthy controls. We further demonstrated the modularity of this platform by successfully transferring it into <i>Pseudomonas koreensis</i> D26, a strain known for its plant growth-promoting properties, indicating broad applicability across biocontrol-relevant pseudomonads. This work establishes a versatile T3SS-based delivery platform for precision biocontrol, offering a generalizable strategy for engineering beneficial rhizobacteria.</p>","PeriodicalId":209,"journal":{"name":"Microbial Biotechnology","volume":"19 6","pages":""},"PeriodicalIF":6.7,"publicationDate":"2026-06-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/1751-7915.70403","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148306412","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Adaptive Laboratory Evolution Unlocks Membrane Permeability as a Key Limitation in Long-Chain Alcohol Metabolism by Pseudomonas putida KT2440","authors":"Raul Mireles,&nbsp;Lianet Noda-García","doi":"10.1111/1751-7915.70399","DOIUrl":"10.1111/1751-7915.70399","url":null,"abstract":"<p><i>Pseudomonas putida</i> KT2440, renowned for its diverse metabolic capabilities, is a promising platform for downstream processing and revalorisation of recalcitrant molecules. In this study, we examined and optimised <i>P. putida</i> KT2440's ability to utilise products of the degradation of polyethylene (PE), the most used and disposed plastic. PE degradation creates over 200 molecules that vary in oxidation level and, thus, chemical properties. Among those, long-chain alcohols represent one of the most challenging fractions to process due to their poor solubility. Using them as feedstock for microbial growth would close the plastic-derived carbon cycle, reducing environmental impact. First, we discovered that <i>P. putida</i> KT2440 can use the long-chain alcohols, 1-hexadecanol and 1-eicosanol, as the sole carbon and energy source. Using adaptive laboratory evolution (ALE), we generated variants with improved growth rates on such substrates. Mutations that became fixed during ALE provided insights into the mechanism, highlighting the importance of cell–substrate interaction. By heterologously expressing a hydrocarbon transporter-encoding gene, we successfully reproduced the ALE-derived phenotype, suggesting that the bottleneck in long-chain alcohol utilisation lies in uptake rather than substrate transformation. These findings lay the groundwork for the potential application of <i>P. putida</i> KT2440 for the valorisation of PE degradation products.</p>","PeriodicalId":209,"journal":{"name":"Microbial Biotechnology","volume":"19 6","pages":""},"PeriodicalIF":6.7,"publicationDate":"2026-06-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/1751-7915.70399","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148306336","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Correction to ‘Unveiling the Potential of Lentilactobacillus hilgardii in Malolactic Fermentation: Comparative Genomics and Fermentation Dynamics’","authors":"","doi":"10.1111/1751-7915.70402","DOIUrl":"10.1111/1751-7915.70402","url":null,"abstract":"<p>\u0000 <span>Mantegazza, G.</span>, <span>N. Mangieri</span>, <span>E. V. Yazdi</span>, <span>P. Russo</span>, <span>D. Mora</span>, and <span>G. Gargari</span>. <span>2025</span>. “ <span>Unveiling the Potential of <i>Lentilactobacillus hilgardii</i> in Malolactic Fermentation: Comparative Genomics and Fermentation Dynamics</span>.” <i>Microbial Biotechnology</i> <span>18</span>, no. <span>12</span>: e70259. https://doi.org/10.1111/1751-7915.70259.\u0000 </p><p>FIGURE 5 <b>CORRECT</b> | Fructose concentration (g/L) at Day 0 and Day 90 during fermentation with <i>Lentilactobacillus hilgardii</i> and <i>Oenococcus oeni</i> at 25°C and 10°C. Data are presented as mean ± standard deviation (<i>n</i> = 3). <i>L. hilgardii</i> exhibited a markedly higher capacity for fructose consumption under both thermal conditions, whereas <i>O. oeni</i> showed limited or no utilisation, particularly at low temperature.</p><p>FIGURE 5 <b>INCORRECT</b>.</p><p>We apologize for this error.</p>","PeriodicalId":209,"journal":{"name":"Microbial Biotechnology","volume":"19 6","pages":""},"PeriodicalIF":6.7,"publicationDate":"2026-06-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/1751-7915.70402","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148275235","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"NADPH Dehydrogenase Gene Regulates Energy Distribution and Fatty Acid Metabolism During Fruiting Body Formation in the Filamentous Fungus Podospora anserina","authors":"Yanling Qiu,&nbsp;Amechi S. Nwankwegu,&nbsp;Yue Sun,&nbsp;Yuanjing Li,&nbsp;Weizhao Chen,&nbsp;Gang Liu,&nbsp;Ning Xie","doi":"10.1111/1751-7915.70401","DOIUrl":"10.1111/1751-7915.70401","url":null,"abstract":"<p>Nicotinamide adenine dinucleotide phosphate (NADPH) dehydrogenase is an oxidoreductase involved in many physiological processes and metabolic pathways. However, its role in filamentous fungal physiology is still unclear. In the present study, three canonical NADPH dehydrogenase genes (<i>Panph1</i>, <i>Panph2,</i> and <i>Panph3</i>) in the fungus <i>Podospora anserina</i> were deleted, and multiple mutants were constructed. Results show a significantly increased number of fruiting bodies in the NADPH dehydrogenase mutant, with the <i>nph</i>^{<i>ΔΔΔ</i>} triple mutant exhibiting higher sensitivity to oxidative stress, suggesting an active-site protein misfolding. Specifically, the antioxidant genes Nox and CAT in the WT were significantly down-regulated, confirming NADPH availability; however, in the NADPH mutant, these genes were significantly upregulated (<i>p</i> ≤ 0.001) as a response to nullify the constraint imposed by NADPH deletion and alleviate oxidative stress. Furthermore, an increase in substrate-level phosphorylation compensated for a significant decrease in oxidative phosphorylation due to NADPH gene deletion. The NADPH/NADP⁺ ratio, a driving force for the intracellular redox potential, showed a significant increase in the <i>nph</i>^{<i>ΔΔΔ</i>} triple mutant. Meanwhile, the NADPH mutant inhibited the <i>β</i>-oxidative pathway, decreasing fatty acid degradation, but promoted fatty acid biosynthesis, reflecting the role of NADPH in the metabolic programming of cellular respiration and energy utilization processes in fungi. Our study provides genetic evidence for the role of the NADPH dehydrogenase gene in oxidative defence and energy metabolism in <i>P. anserina</i>.</p>","PeriodicalId":209,"journal":{"name":"Microbial Biotechnology","volume":"19 6","pages":""},"PeriodicalIF":6.7,"publicationDate":"2026-06-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/1751-7915.70401","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148256716","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}