Enzyme and Microbial Technology最新文献

{"title":"Improved total carotenoid content from Planococcus plakortidis NIOT3 through microwave assisted extraction and genome-guided pathway annotation","authors":"Nishanthika Thenmozhi Kulasekaran,&nbsp;Mary Leema J T,&nbsp;Vishal Vasavan M V,&nbsp;Dharani Gopal,&nbsp;Jeya Marimuthu","doi":"10.1016/j.enzmictec.2025.110707","DOIUrl":"10.1016/j.enzmictec.2025.110707","url":null,"abstract":"<div><div>With an increasing demand for natural colorants, marine microbes have become attractive targets for novel natural colorants like carotenoids. Microbial carotenoid extraction by green approach is advantageous over traditional methods which minimize energy usage and reduce extraction time. In the current study, an orange pigmented marine bacterium <em>Planococcus plakortidis</em> NIOT3 was isolated from the Sesostris bank of Arabian Sea and found to produce desirable level of total carotenoids (320 ± 24 µg/g DW) when grown in rich medium. An energy efficient green approach using Microwave Assisted Extraction was evaluated for carotenoid yield by one factor and statistical optimization of the MAE process conditions including exposure time, alkali concentration and Solid:Liquid ratio which resulted in a 3.26 fold increase in total carotenoid content of 2835 ± 152 µg/g DW. Genome sequencing and annotation revealed genes that are involved in C30 carotenoid biosynthesis. C30 carotenoids like glycosylated diapolycopene and methyl-5-glucosyl-5,6-dihydro-apo-4,4’-lycopenoate produced by NIOT3 were identified by LC-MS analyses as major products. Additionally, LC-MS analysis revealed a minor product with a molecular weight of <em>m/z</em> of 568.84 which corresponds to zeaxanthin, a C40 carotenoid. The study highlights the biotechnological potential of <em>P. plakortidis</em> in carotenoid production.</div></div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"190 ","pages":"Article 110707"},"PeriodicalIF":3.4,"publicationDate":"2025-07-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144570332","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":"2025-07-02","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}

{"title":"Structural determinants of unique substrate specificity of d-amino acid oxidase of the thermophilic fungus Rasamsonia emersonii","authors":"Yuya Shimekake,&nbsp;Takehiro Furuichi,&nbsp;Daiki Imanishi,&nbsp;Shouji Takahashi","doi":"10.1016/j.enzmictec.2025.110705","DOIUrl":"10.1016/j.enzmictec.2025.110705","url":null,"abstract":"<div><div><span>d</span>-Amino acid oxidase from the thermophilic fungus <em>Rasamsonia emersonii</em> (<em>Re</em>DAAO) has garnered attention due to its high stability and broad substrate specificity, making it a promising candidate for various applications. In this study, we explored the structural factors underlying the unique substrate specificity of <em>Re</em>DAAO, particularly its broad substrate range and <span>d</span>-Glu oxidation ability. Comparing <em>Re</em>DAAO with <em>Td</em>DAAO—a homologous <span>d</span>-amino acid oxidase from the thermophilic fungus <em>Thermomyces dupontii</em>—revealed that <em>Re</em>DAAO lacks the YVLQG loop present in <em>Td</em>DAAO, which exhibited narrower substrate specificity. Inserting the YVLQG loop into <em>Re</em>DAAO narrowed its substrate specificity to match <em>Td</em>DAAO, while deleting the sequence from <em>Td</em>DAAO broadened its substrate specificity, resembling <em>Re</em>DAAO. A <em>Td</em>DAAO structural model suggests that the YVLQG loop could interact with a spatially adjacent region covering the active site, distinct from the canonical active-site lid in DAAOs, creating steric hindrance that limits access to the catalytic pocket. Additionally, the unexpected activity of <em>Re</em>DAAO toward <span>d</span>-Glu appears to depend on Arg97 and Ser231, which could interact with <span>d</span>-Glu side chain. Alanine substitutions at these residues significantly reduced <span>d</span>-Glu activity, revealing that Arg97 is essential for catalytic turnover while Ser231 is critical for substrate binding. Together, these results suggest that the YVLQG loop together with the spatially adjacent region acts as a steric gate that modulates access to the catalytic pocket, and Arg97/Ser231 plays an important role in <span>d</span>-Glu. These findings deepen our understanding of the structure–function relationship of DAAO and provide a foundation for developing improved DAAO variants for industrial applications.</div></div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"190 ","pages":"Article 110705"},"PeriodicalIF":3.4,"publicationDate":"2025-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144557435","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":"Optimization of physico-chemical parameters for synergistic production of xylano-cellulolytic enzymes by novel Pantoea sp. (PQ584882) under solid-state fermentation using statistical design approach","authors":"Neha Maurya ,&nbsp;Harsh Sable ,&nbsp;Jyoti Chauhan ,&nbsp;Amit Kumar ,&nbsp;Sharad Agrawal","doi":"10.1016/j.enzmictec.2025.110697","DOIUrl":"10.1016/j.enzmictec.2025.110697","url":null,"abstract":"<div><div>Xylanase and cellulase have become increasingly significant due to their versatile applications in the food, paper, biofuel, and pharmaceutical industries. Nevertheless, the current production of these enzymes relies on costly substrates, with estimates indicating that over 30 % of the production expenses are attributed to these substrates. The objective of this study is to optimize the physicochemical parameters for obtaining the maximum production of xylanase &amp; cellulase enzyme from <em>Pantoea sp.</em> (PQ584882). The production conditions were statistically optimized using Plackett-Burman design (PBD) and Central Composite design (CCD). The significant variables identified through PB design including temperature, substrate-to-moisture ratio, K₂HPO₄, peptone, surfactant, inoculum size, inoculum age, incubation time, and pH were further optimized using the CCD approach. This optimization process revealed the most influential factors affecting xylanase &amp; cellulase production, with optimal conditions observed at a temperature of 40◦C, Moisture Proportion, 15 mL; K₂HPO₄ 6 mM; peptone, 1.55 %; Castor oil 0.5 %; inoculum size, 1.55 % (v/w); inoculum age, 18 h; an incubation time, 87 h. The optimized CCD model displayed a 1.84-fold greater xylanase &amp; cellulose production than the PB design approach. These findings suggest that wheat bran, a readily available agro-waste, could be a feasible alternative to the conventional substrate, beechwood xylan and CMC (Carboxy methyl cellulose) for the production of xylanase &amp; cellulase enzymes with the possibility of achieving higher production levels optimized by using a statistical design approach<strong>.</strong></div></div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"190 ","pages":"Article 110697"},"PeriodicalIF":3.4,"publicationDate":"2025-06-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144489382","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":"Glycosylation of terpenes with a substrate-flexible recombinant glycosyltransferase from Micromonospora inoyensis NRRL 3292","authors":"Mingyu Yeo ,&nbsp;Bohyun Choi ,&nbsp;Hong Ji Im ,&nbsp;Jiwon Na ,&nbsp;Siyeon Choi ,&nbsp;Hyunsung So ,&nbsp;Je Won Park","doi":"10.1016/j.enzmictec.2025.110704","DOIUrl":"10.1016/j.enzmictec.2025.110704","url":null,"abstract":"<div><div>Glycosyltransferase (GT)-specific degenerate PCR screening of the established fosmid libraries of the soil actinomycete <em>Micromonospora inoyensis</em> NRRL 3292, followed by <em>in silico</em> sequencing of target clones, allowed us to isolate members of the family 1 GT-encoding gene. A recombinant MiTGT, as a His-tagged protein, was heterologously expressed in <em>Escherichia coli</em>. Its bio-catalytic reactions with both natural terpene aglycones derived from natural sweeteners including stevioside and mogroside (as glycosyl acceptors) and nucleotide-activated hexoses (as glycosyl donors) created a number of structurally diversified terpene glycosides, thus characterizing MiTGT as a terpene glycosyltransferase with substrate-flexibility. Chromatographic isolation of the product glycosides followed by the instrumental analyses, clearly confirmed the previously unprecedented stereospecific glycosides as steviol-13-<em>O</em>-α-glucoside, steviol-13-<em>O</em>-α-(2′-deoxy)glucoside and steviol-13-<em>O</em>-α-galactoside, mogrol-3-<em>O</em>-α-glucoside, mogrol-3-<em>O</em>-α-(2′-deoxy)glucoside, mogrol-3-<em>O</em>-α-galactoside. Moreover, their elastase inhibitory activities together with anti-proliferative activities against the human pancreatic adenocarcinoma cell line PANC-1 were further investigated, hence representing the cosmeceutical and pharmaceutical potentials of the designated terpene glycosides. This is the first report on the microbe-origin bio-catalytic production of unnatural terpene α-configured glycosides.</div></div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"190 ","pages":"Article 110704"},"PeriodicalIF":3.4,"publicationDate":"2025-06-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144502061","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":"Microbial community dynamics and functional potential during the natural fermentation of rose: A metagenomic and volatile compound analysis","authors":"Zhiyuan Yin,&nbsp;Kangdi Cao,&nbsp;Ningfei Duan,&nbsp;Zhiguo Zhang","doi":"10.1016/j.enzmictec.2025.110703","DOIUrl":"10.1016/j.enzmictec.2025.110703","url":null,"abstract":"<div><div>This study investigates the dynamics of microbial communities and their functional characteristics during the natural fermentation of roses. Utilizing metagenomic sequencing and volatile compound analysis, the research elucidates the succession of microbial communities and their relationship with the flavor compound production. The findings indicate that <em>Klebsiella</em> and <em>Pichia</em> are predominant in the early stages of fermentation, while <em>Acetobacter</em> and <em>Cyberlindnera</em> become more abundant in the middle and later stages. The glycosyltransferase (GT) family is identified as the primary carbohydrate-active enzyme (CAZy) family involved in fermentation, with GT1 and GT2 exhibiting a higher gene abundance. Functional genes are predominantly associated with the carbohydrate and amino acid metabolism. Analysis of volatile compounds reveals that substances such as phenethyl acetate and (S,S)-2,3-Butanediol are closely related to the structure of the microbial community. These findings contribute to a deeper understanding of the mechanisms underlying rose fermentation and offer a theoretical foundation for technological advancements in the rose product industry.</div></div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"190 ","pages":"Article 110703"},"PeriodicalIF":3.4,"publicationDate":"2025-06-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144481156","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":"Crystal structure of cyclohexylamine oxidase from Acinetobacter sp. YT−02 reveals key residues for catalytic activity and substrate specificity","authors":"Jing Wu ,&nbsp;Zhenggang Han ,&nbsp;Pengrong Li ,&nbsp;Jing Li ,&nbsp;Yuanyuan Chen ,&nbsp;Shangbo Ning ,&nbsp;Hong-jun Chao ,&nbsp;Xue-wang Gao ,&nbsp;Dazhong Yan","doi":"10.1016/j.enzmictec.2025.110700","DOIUrl":"10.1016/j.enzmictec.2025.110700","url":null,"abstract":"<div><div>Cyclohexylamine oxidase is a member of amine oxidases that catalyzes the conversion of cyclohexylamine to cyclohexanone. In our previous work, the enzymatic activity assay of cyclohexylamine oxidase CHAO_YT-02 indicated that its specific activity towards cyclohexylamine of CHAO_YT-02 was ten times higher than that of its homolog CHAO_IH-35A. In this study, the crystal structure of CHAO_YT-02 was determined by the molecular replacement method at a resolution of 1.49 Å. The atomic structure revealed that the amino acid residues Leu302, Trp70, Phe197, Phe349, and Tyr440 constitute the active center pocket of the enzyme. Amino acid residues Ile180, Leu181, and Trp332 separate the active center pocket and an intermediate pocket. Moreover, a molecular dynamics (MD) simulation and the calculation of the binding free energy were performed to predict substrate entry and product release from cyclohexylamine oxidases. Single-amino acid substitution mutants (W70A, I180A, L181A, I208A, F197A, L302A, W332A, F349A, and Y440A) of CHAO_YT-02 were constructed to investigate the role of these amino acid residues in enzymatic properties and substrate specificity. The results indicated that both the amino acid residues in the active center pocket and gating the two pockets affected the activity or substrate specificity of CHAO_YT-02. This study on the structure and catalytic mechanism of cyclohexylamine oxidase is beneficial to eliminating toxic amine compounds in the environment.</div></div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"190 ","pages":"Article 110700"},"PeriodicalIF":3.4,"publicationDate":"2025-06-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144549870","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":"The effect of the cspA gene on growth development and butenyl-spinosyn biosynthesis in Saccharopolyspora pogona","authors":"Duo Jin,&nbsp;Shanrui Wang,&nbsp;Wangqiong Chen,&nbsp;Jing Fang,&nbsp;Jie Rang,&nbsp;Liqiu Xia,&nbsp;zirong zhu","doi":"10.1016/j.enzmictec.2025.110701","DOIUrl":"10.1016/j.enzmictec.2025.110701","url":null,"abstract":"<div><div>Cold shock proteins (CSPs) represent a universal class of proteins in microorganisms, rapidly inducible under low temperature conditions. As molecular chaperones for RNA, they bind to single-stranded nucleotides, preventing the formation of complex secondary structures. This facilitates efficient translation and gene expression regulation. This investigation pioneers the study of the <em>cspA</em> gene through metabolic engineering techniques, to uncover its critical biological roles in the growth and development of <em>Saccharopolyspora pogona</em> and in butenyl-spinosyn biosynthesis. Employing comparative proteomic and targeted metabolomic analyses, this research elucidates the metabolic pathway alterations prompted by the augmented presence of the cold shock protein CspA. Additionally, it offers initial insights into the regulatory mechanisms by which CspA affects <em>S. pogona</em>'s growth, development, and butenyl-spinosyn production. The outcomes of this study significantly advance our theoretical understanding of the rational optimization of butenyl-spinosyn biosynthetic pathways. They also provide valuable guidance for other actinobacteria aiming to boost their resilience to harsh environments by overexpressing the <em>cspA</em> gene.</div></div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"190 ","pages":"Article 110701"},"PeriodicalIF":3.4,"publicationDate":"2025-06-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144471575","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":"Deciphering key residues governing thermostability in Thermomyces lanuginosus lipase through Gibbs free energy-guided engineering","authors":"Xia Xiang ,&nbsp;Sidi Wan ,&nbsp;Songjing Zhang ,&nbsp;Enheng Zhu ,&nbsp;Xuejun Lin ,&nbsp;Nanyu Han","doi":"10.1016/j.enzmictec.2025.110702","DOIUrl":"10.1016/j.enzmictec.2025.110702","url":null,"abstract":"<div><div>Lipases serve as indispensable biocatalysts in many industrial applications due to their versatile catalytic abilities. To ensure their thermal resilience of the harsh biological treatment in industry, it is crucial to identify key residues which might impact thermostability. Here, computational design was adopted to decode the stability-determining residues in <em>Thermomyces lanuginosus</em> lipase (TLL). Systematic Gibbs free energy profiling of potent TLL single-point mutational candidates predicted proline 256 (P256) as a thermal liability hotspot. Saturation mutagenesis at P256 discovered that among nineteen P256 variants: (1) five P256 variants exhibited increased melting temperature (ΔT_m up to 2°C); (2) six variants displayed an optimum temperature with 5–10°C elevation; (3) five P256 variants retained up to 21 % higher residual activity after incubation at 80°C. Furthermore, both P256E and P256I demonstrated synergistic improvements in biodiesel conversion rates, P256I specifically exhibited long-term and recycling stability. Molecular dynamics simulations revealed that the substitutions in P256A/E/I/K with compensatory main-chain rotational freedom, facilitating hydrogen bonding network with both upstream and downstream residues, thereby preserving local structural stability. This study pioneers the identification of P256 as a critical residue governing TLL thermostability. Furthermore, our Gibbs-guided engineering strategy generates multi-property-enhanced lipase variants, directly addressing industrial demands.</div></div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"190 ","pages":"Article 110702"},"PeriodicalIF":3.4,"publicationDate":"2025-06-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144471581","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":"Bacillus cereus N-acyl homoserine lactonase and penicillin acylase II against Pseudomonas aeruginosa: An In silico and In vitro investigations exploring the effects of gamma radiation on their quorum quenching activity","authors":"Radwa N. Morgan,&nbsp;Reham R. El-Behery","doi":"10.1016/j.enzmictec.2025.110699","DOIUrl":"10.1016/j.enzmictec.2025.110699","url":null,"abstract":"<div><div>This study explored the quorum quenching (QQ) activities of ISM25 strain, sourced from an environmental setting, on pathogenic <em>Pseudomonas aeruginosa</em> isolates. The isolate ISM25 was first screened for the presence of both <em>AiiA</em> acyl homoserine lactonase and penicillin <em>acylase II</em> genes. The in silico investigation focused on examining the physiochemical properties and QQ activities in relation to both short (&lt;C8) and long chain (&gt;C12) homoserine lactones (HSLs) utilizing AutoDock Vina docking tests. Potential interactions between the acylase II enzyme and carbapenem antibiotics were also investigated during the in silico docking tests. Afterwards, the in vitro QQ activity of ISM25 crude protein extract was assessed against the biofilm index, AHLs molecules, and meropenem minimum inhibitory concentrations of pathogenic <em>P. aeruginosa</em> isolates. The findings from the in silico docking analysis were supported by the crude protein extract's capacity to hydrolyze AHLs generated by the pathogenic <em>P. aeruginosa</em> isolate, as well as the reference C12-HSL signal molecule. The notable decrease in biofilm indices (P &lt; 0.05) following exposure to the crude ISM25 extract further corroborated the expression of both QQ enzymes by the ISM25 isolate and their in silico QQ activity. The crude extract from ISM25 lowered the MIC and sensitized <em>P. aeruginosa</em> to meropenem (P &lt; 0.05), suggesting that ISM25 crude extract containing acylase II can be administered in conjunction with meropenem without affecting its efficacy. Finally, exposure of ISM25 to gamma radiation did not impair its QQ activity at doses below 100 Gy, however, QQ activity was nearly abolished at radiation dose ≥ 500 Gy.</div></div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"190 ","pages":"Article 110699"},"PeriodicalIF":3.4,"publicationDate":"2025-06-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144365395","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}