Journal of Industrial Microbiology & Biotechnology最新文献

{"title":"Exploring the compatibility of phosphopantetheinyl transferases with acyl carrier proteins spanning type II polyketide synthase sequence space.","authors":"Areta L N Bifendeh, Kenneth K Hsu, Christina M McBride, Charlie M Ferguson, Eva R Baumann, Diego Capcha-Rodriguez, Xinnuo Chen, Berlensie Chery, Margo M Chihade, Paola Delgado Umpierre, Taliyah Evans, Carolyn H Everett, Syeda F Faheem, Oscar D Garrett, Aliya R Gottesfeld, Ishir G Gupta, Jason D Haas, Theresa A Haupt, Jean Katz, Sadie Kim, Matthias Langer, Vy Le, Kevin K Li, Baldwin Zhao, Siyue Lin, Kelsey N Mabry, Anna Malkov, Abigail T Marquis, Kieran R McDonnell, Kristen Min, Nicholas B Mostaghim, Krysta M Nichols, Rebecca A Osbaldeston, Trisha T Phan, Alana T Ponte, Tala Qaraqe, Bianca S Rosas, Caroline S Smith, Logan E Smith, Maisie W Smith, Aviva C R Soll, Gabriel Rocco Sotero, Isabel E Thornberry, Kristina Tran, Quynh K Vo, Marcos G Yoc-Bautista, Madison Young, Kelly A Zukowski, Robert Fairman, Kimberly A Wodzanowski, Michael A Herrera, Yae In Cho, Louise K Charkoudian","doi":"10.1093/jimb/kuaf031","DOIUrl":"https://doi.org/10.1093/jimb/kuaf031","url":null,"abstract":"<p><p>Phosphopantetheinyl transferases (PPTases) play an essential role in primary and secondary metabolism. These enzymes facilitate the post-translational activation of acyl carrier proteins (ACPs) central to the biosynthesis of fatty acids and polyketides. Modulation of ACP-PPTase interactions is a promising approach to both increase access to desired molecular outputs and disrupt mechanisms associated with disease progression. However, such an approach requires understanding the molecular principles that govern ACP-PPTase interactions across diverse synthases. Through a multi-year, course-based undergraduate research experience (CURE), 17 ACPs representing a range of putative type II polyketide synthases, from actinobacterial and non-actinobacterial phyla, were evaluated as substrates for three PPTases (AcpS, Sfp, and vulPPT). The observed PPTase compatibility, sequence-level analyses, and predictive structural modelling suggest that ACP selectivity is driven by amino acids surrounding the conserved, modified serine on the ACP. We propose that vulPPT and Sfp are driven primarily by hydrophobic contacts, whereas AcpS may favor ACPs which exhibit high net-negative charge density, as well as a broad electronegative surface distribution. Furthermore, we report a plausible, hitherto unreported hydrophobic interaction between vulPPT and a conserved ACP crease, upstream of the invariant serine, which may facilitate docking. This work provides a catalog of compatible and incompatible ACP-PPTase partnerships, highlighting specific regions on the ACP and/or PPTase that show promise for future strategic engineering and inhibitor development efforts.</p>","PeriodicalId":16092,"journal":{"name":"Journal of Industrial Microbiology & Biotechnology","volume":" ","pages":""},"PeriodicalIF":3.2,"publicationDate":"2025-09-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145176163","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Finding of the positive impact of glucose on the production of indican over indigo in engineered Escherichia coli.","authors":"Hyun Jin Kim, Yeda Lee, Yuni Shin, Suhye Choi, Jinok Oh, Suwon Kim, Jungoh Ahn, Kwon-Young Choi, Jeong Chan Joo, Shashi Kant Bhatia, Yung-Hun Yang","doi":"10.1093/jimb/kuae048","DOIUrl":"10.1093/jimb/kuae048","url":null,"abstract":"<p><p>Indigo is a plant-based natural blue dye that can be produced via chemical synthesis and biological pathways. However, the toxic reduction processes and intracellular production of indigo through microbial metabolism are often limited by insolubility of indigo and complex downstream processing, causing environmental issues in the dyeing processes. Additionally, indican, a precursor of indigo with a glucose moiety, is highly soluble and can be easily converted into indoxyl by β-glucosidase, forming indigo under mild conditions. We constructed an indican-producing strain Escherichia coli BL21 HI201 by introducing a UDP-glycosyltransferase (ugt) into an indoxyl production system containing tryptophanse (tnaA) and flavin-containing monooxygenase (FMO) genes, enabling conversion of tryptophan into indican. Testing of the effect by various carbon sources suggested that glucose is one of the major factors affecting the ratio of indigo to indican, and increase in glucose concentration to more than 1.5% could produce sole indican without indigo. Under optimal conditions, E. coli BL21 HI201 biosynthesized 5.65 mM indican from tryptophan. Additionally, after deletion of various β-glucosidase genes, the bglA knockout strain E. coli BL21 HI204 produced more indican, achieving 6.79 mM after 24 hr of cultivation. This study demonstrated the strategic production of indican through the installation of a production system, deletion of a byproduct pathway, and control of glucose concentration.</p><p><strong>One-sentence summary: </strong>This paper demonstrates the strategic enhancement of indican production in genetically engineered Escherichia coli BL21 by optimizing metabolic pathways and controlling glucose concentrations.</p>","PeriodicalId":16092,"journal":{"name":"Journal of Industrial Microbiology & Biotechnology","volume":"52 ","pages":""},"PeriodicalIF":3.2,"publicationDate":"2024-12-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11947661/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143730374","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Enhanced amino acid biosynthesis in Phaffia rhodozyma via herbicide-induced selection.","authors":"Svetlana Raita, Iveta Kuzmika, Taras Mika, Zane Geiba, Kriss Spalvins","doi":"10.1093/jimb/kuaf011","DOIUrl":"10.1093/jimb/kuaf011","url":null,"abstract":"<p><p>According to the Food and Agricultural Organisation 2024 statement, developing single-cell protein technology is important to reduce the burden on conventional feed protein production sectors. In this regard, improved commercial strains rich in amino acids, especially Lys and Met, may provide a sustainable alternative source of protein in aquaculture diets. The developed and laboratory-validated methodology for creating protein-synthesizing mutants will strengthen the competitiveness of SCP production technology. The present work provides unique results on improving the protein-producing properties of wild-type Phaffia rhodozyma DSM 5626 by mutagenesis and screening on herbicide-containing medium as a selective agent for amino acid biosynthesis inhibition. Inhibitory concentrations of pure herbicide actives were determined for S-(2-aminoethyl)-L-cysteine (AEC) hydrochloride and glufosinate-ammonium (GA) for complete inhibition and strong inhibition of the DSM 5626 strain. GA at a concentration of 50 mM and 100 mM and AEC at 0.5 mM and 2.5 mM were chosen for mutant selection after chemical mutagenesis. The use of herbicides resulted in the selection of mutants with significantly improved synthesis of Met and Lys. Specifically, mutants GA6/4 and GA7/5 exhibited 37% and 26% higher Met levels, respectively, while GA6/3 had a 14% increase in Lys compared to the wild-type strain. The AEC3/9 mutant demonstrated a 35% increase in Met, 24% in Lys, 8% in Ile, and 6% in Phe, underscoring the efficacy of this screening approach in enhancing essential amino acid content. The protein quality parameters essential amino acid index and amino acid score of these mutants became higher compared with commercial strains of SCP yeast such as C. utilis, S. cerevisiae, K. marxianus, etc.</p><p><strong>One-sentence summary: </strong>Mutagenesis combined with selective screening using herbicides is an effective approach to enhancing amino acid biosynthesis in yeast.</p>","PeriodicalId":16092,"journal":{"name":"Journal of Industrial Microbiology & Biotechnology","volume":" ","pages":""},"PeriodicalIF":3.2,"publicationDate":"2024-12-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12107244/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144021174","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Production of extracellular L-arginase by Alcaligenes aquatilis BC2 isolated from soda lakes (Lake Chitu) of Ethiopia.","authors":"Birhan Getie Assega, Kefyalew Ayalew Getahun, Tamene Milkessa, Tsehayneh Geremew Yohannes, Feleke Moges, Mulugeta Aemero, Berhanu Andualem","doi":"10.1093/jimb/kuaf017","DOIUrl":"10.1093/jimb/kuaf017","url":null,"abstract":"<p><p>L-Arginase is a therapeutic enzyme that hydrolyzes L-arginine to ornithine and urea. The L-arginase extracted from bacteria has an anticancer activity by causing starvation of nutrients for cancer cells. This study aimed to screen and characterize L-arginase-producing bacteria and to optimize different factors influencing L-arginase production. Isolation and primary screening were carried out by using mineral arginine agar media using phenol red as an indicator. Molecular identification of the isolates was employed by using 16S ribosomal RNA sequencing and phylogenetic tree construction. L-Arginase assay by colorimetric method was carried out to measure the amount of urea liberated from the hydrolysis of L-arginine for quantitative screening. From 31 water samples, 102 colonies were isolated, and those colonies that convert the media to pink were selected as arginase-producing bacteria. 7 isolates were screened from qualitative screening method. Based on quantitative screening, the highest L-arginase was produced from bacteria Alcaligenes aquatilis BC2 (92.46 ± 0.19 U/ml) followed by Paenalcaligenes suwonensis BCW8 (59.29 ± 0.66 U/ml). Following their mean difference, isolate BC2 was selected for further optimization process of 8 parameters. After optimization, the isolate shows the maximum (163.85 U/ml) enzyme activity. The result of this study implies that novel bacteria were isolated from soda lakes that produce a considerable amount of L-arginase, which can be used as a promising anticancer activity. One-Sentence Summary: This study successfully isolated and characterized a novel L-arginase-producing bacterium, Alcaligenes aquatilis BC2, from Ethiopian soda lakes and optimized its enzyme production parameters for potential anticancer applications.</p>","PeriodicalId":16092,"journal":{"name":"Journal of Industrial Microbiology & Biotechnology","volume":" ","pages":""},"PeriodicalIF":3.2,"publicationDate":"2024-12-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12259280/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144505960","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Genome-mining-guided discovery of coumarubrin: A novel aminocoumarin-substituted rubromycin antibiotic.","authors":"Heiner G Weddeling, Sven T Sowa, Selina Bosshardt, Lukas Schwimbersky, Malik Rakhmanov, Robin Teufel","doi":"10.1093/jimb/kuaf018","DOIUrl":"10.1093/jimb/kuaf018","url":null,"abstract":"<p><p>Rubromycins are bacterial aromatic polyketides containing a hallmark spiroketal pharmacophore produced by type II polyketide synthases and accessory enzymes. They generally display cytotoxic and antimicrobial properties, frequently disrupting cellular processes and proteins associated with nucleic acids, such as DNA helicase or telomerase. Among the known rubromycin congeners, hyaluromycin stands out due to a 2-amino-3-hydroxycyclopent-2-enone (C5N) substitution that is presumably installed by an amide bond synthetase (ABS). Here, we used bioinformatic analysis to identify uncharacterized biosynthetic gene clusters and potential rubromycin producer strains encoding putative ABSs but lacking the enzymes responsible for C5N formation, suggesting potentially novel substituents. One of these strains, Lentzea tibetensis, was successfully cultivated and confirmed to produce a previously undescribed aminocoumarin-substituted rubromycin polyketide, named coumarubrin, as verified by high-resolution mass spectrometry (HRMS) and comprehensive nuclear magnetic resonance (NMR) spectroscopy. Electronic circular dichroism spectroscopy indicates an absolute configuration identical to that of previously characterized rubromycins, while the first bioactivity assays demonstrated potent inhibitory activity against Gram-positive bacteria. One-Sentence Summary: This study reports the discovery of a novel member of the rubromycins, antibiotic and cytotoxic aromatic polyketides produced by Actinobacteria, which is fused to a distinct aminocoumarin moiety.</p>","PeriodicalId":16092,"journal":{"name":"Journal of Industrial Microbiology & Biotechnology","volume":" ","pages":""},"PeriodicalIF":3.2,"publicationDate":"2024-12-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12284476/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144560282","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Recent advancement of glucosamine and N-acetyl glucosamine production using microorganisms: A review.","authors":"Anica Tasnim Protity, Shengde Zhou","doi":"10.1093/jimb/kuaf014","DOIUrl":"10.1093/jimb/kuaf014","url":null,"abstract":"<p><p>Glucosamine (GlcN) and GlcN-based supplements, e.g. glucosamine hydrochloride, glucosamine sulfate, and N-acetyl glucosamine (GlcNAc), provide symptomatic relief to osteoarthritis patients and have been used as one of the most popular nutraceuticals. To meet the increasing demands, scientists have explored cost-effective methods for GlcN and GlcNAc production using low-cost raw materials such as seafood waste. However, the commercially available GlcN and GlcNAc production methods are environmentally harmful because of the use of toxic reagents. Moreover, the raw material used might be unsafe for consumers with shrimp allergies. On the other hand, bio-based GlcN production is gaining popularity because of its eco-friendly production approach and optimum reaction conditions. In this mini-review, we will discuss the recent developments to produce GlcN and GlcNAc through (1) the chemical and enzyme-mediated approaches of crude chitin hydrolysis, primarily obtained from shrimp and crabs; (2) the whole cell-based systems for fungal derived chitin bio-transformation and fungal fermentation; and (3) the metabolic engineering and the adaptive evolution based microbial biocatalyst for a balanced cell growth and optimal production of GlcN and GlcNAc. One-Sentence Summary: This article summarizes the mechanism of glucosamine and N-acetyl glucosamine production using bacteria, fungi, and chemical processes.</p>","PeriodicalId":16092,"journal":{"name":"Journal of Industrial Microbiology & Biotechnology","volume":" ","pages":""},"PeriodicalIF":3.2,"publicationDate":"2024-12-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12168751/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144159638","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Biosynthesis of guanidine-containing natural products in cyanobacteria.","authors":"Wenhe Zhang, Richiro Ushimaru","doi":"10.1093/jimb/kuaf024","DOIUrl":"10.1093/jimb/kuaf024","url":null,"abstract":"<p><p>Cyanobacteria are prolific producers of structurally diverse and biologically potent natural products, a subset of which feature guanidino moieties. Introduction and modification of the guanidine group confer tuned basicity and enable extensive hydrogen bonding, cation-π, and electrostatic interactions, facilitating high-affinity binding to numerous biological targets. Although the enzymatic processes responsible for guanidine modifications in cyanobacterial pathways remain somewhat obscure, recent investigations have begun to clarify the biosynthetic machinery that mediates these distinctive transformations. In this review, we summarize these advances, with particular emphasis on the enzymatic steps responsible for guanidine installation and tailoring. These enzymatic transformations include N-prenylation, cyclization, and tricyclic guanidinium formation, representing rare or previously undescribed biosynthetic strategies in nature. This review provides new insights into the metabolic and enzymatic versatility of cyanobacteria and a foundation for future advances in enzyme engineering and therapeutic discovery. One-Sentence Summary: This review highlights recent advances in understanding how cyanobacteria enzymatically install and modify guanidino groups to produce bioactive natural products.</p>","PeriodicalId":16092,"journal":{"name":"Journal of Industrial Microbiology & Biotechnology","volume":" ","pages":""},"PeriodicalIF":3.2,"publicationDate":"2024-12-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12371840/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144753573","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Biocatalytic diversification of abietic acid in Streptomyces.","authors":"Caitlin A McCadden, Tyler A Alsup, Ion Ghiviriga, Jeffrey D Rudolf","doi":"10.1093/jimb/kuaf003","DOIUrl":"10.1093/jimb/kuaf003","url":null,"abstract":"<p><p>Biocatalysis provides access to synthetically challenging molecules and commercially and pharmaceutically relevant natural product analogs while adhering to principles of green chemistry. Cytochromes P450 (P450s) are among the most superlative and versatile oxidative enzymes found in nature and are desired regio- and stereoselective biocatalysts, particularly for structurally complex hydrocarbon skeletons. We used 10 genome-sequenced Streptomyces strains, selected based on their preponderance of P450s, to biotransform the bioactive diterpenoid abietic acid. We isolated and structurally characterized seven oxidized abietic acid derivatives from three different strains, including four products that are new bacterial biotransformants or enzymatic products. Oxidations (hydroxylation, dehydrogenation, and aromatization) were seen on both the B and C rings of abietic acid and five products had multiple modifications. Notable conversions observed in the study were that of abietic acid to 15-hydroxy-7-oxo-8,11,13-abietatrien-18-oic acid, 7, which involves multiple hydroxylation reactions and dehydrogenation. The findings from this study will lead to identifying P450s or other enzymes that may act as general biocatalysts to modify abietanes and other labdane-type diterpenoid skeletons.</p><p><strong>One-sentence summary: </strong>Genome-guided biotransformation of the bioactive diterpenoid abietic acid in Streptomyces yielded seven oxidized derivatives including four that have not been previously seen from bacteria.</p>","PeriodicalId":16092,"journal":{"name":"Journal of Industrial Microbiology & Biotechnology","volume":" ","pages":""},"PeriodicalIF":3.2,"publicationDate":"2024-12-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11812575/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143065574","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"An overview of key industrial product citric acid production by Aspergillus niger and its application.","authors":"Ambreen Latif, Noor Hassan, Hazrat Ali, Muhammad Bilal Khan Niazi, Zaib Jahan, Iqra Latif Ghuman, Farwa Hassan, Anam Saqib","doi":"10.1093/jimb/kuaf007","DOIUrl":"10.1093/jimb/kuaf007","url":null,"abstract":"<p><p>Citric acid possesses high economic value and is considered as the world's largest consumed organic acid in numerous industries. Citric acid applications range from food to beverage industries, pharmaceuticals, cosmetics, and the environment. It is mostly produced by microbial fermentation, but Aspergillus niger is considered as the main workhorse for large-scale production of citric acid. In the current review, special devotion has been made toward addressing the latest and innovative literature related to production of citric acid by A. niger. The review article discusses A. niger historical involvement in citric acid production, fermentation technologies, molecular biology, biosynthesis, accumulation of citric acid, methods for enhanced production of citric acid, different operational factors also influencing citric acid production, and various techniques used for citric acid recovery. Also, copious biotechnological applications of citric acid are summarized for a fundamental comprehension of the subject and its critical role in diverse fields of industries.</p><p><strong>One-sentence summary: </strong>This review describes the historical role of Aspergillus niger in the production of citric acid, fermentation technologies, molecular biology, techniques for increased citric acid production, and other physical and chemical variables influencing the production of citric acid.</p>","PeriodicalId":16092,"journal":{"name":"Journal of Industrial Microbiology & Biotechnology","volume":" ","pages":""},"PeriodicalIF":3.2,"publicationDate":"2024-12-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11956825/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143742898","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Metagenomic mining unveils a novel GH130 enzyme with exclusive xylanase activity over a wide temperature and pH ranges.","authors":"Amr A Hemeda, Sara A Zahran, Marwa Ali-Tammam, Menna A Ewida, Mona T Kashef, Aymen S Yassin, Avishek Mitra, Noha H Youssef, Mostafa S Elshahed","doi":"10.1093/jimb/kuaf006","DOIUrl":"10.1093/jimb/kuaf006","url":null,"abstract":"<p><p>The equine gut harbors a diverse microbial community and represents a rich source of carbohydrate-active enzymes (CAZymes). To identify and characterize potentially novel CAZymes from a horse's hindgut metagenome, shotgun metagenomic sequencing was performed on DNA extracted from a stool sample of a male horse, followed by CAZyme annotation. Here, we report on the characterization of a novel enzyme (AH2) that was identified, synthesized, cloned, and characterized from the obtained CAZyme dataset. AH2 was identified as a GH130 family member and displayed exclusive xylanase activity, a trait hitherto unreported in prior characterization of GH130 CAZymes. AH2 displayed an optimal activity at a pH of 5.6 and a temperature of 50°C. AH2 maintained significant activity across a pH range of 4-10 (62-72%) and temperatures of 30-70°C (77-86%). The enzyme had remarkable stability, with minimal reductions in activity across a temperature range of 4-70°C and pH levels of 3, 7, and 9. Docking studies identified AH2's amino acids (Glu90 and Glu149) to be involved in substrate binding. Molecular dynamics simulation confirmed the structural stability of AH2 at pH 5.6 and 50°C, further supporting its resilience under these conditions. Our results expand on the known activities associated with the GH130 CAZyme family and demonstrate that the horse gut metagenome represents an unexplored source of novel CAZymes.</p><p><strong>One-sentence summary: </strong>A novel activity for members of the CAZyme family GH130.</p>","PeriodicalId":16092,"journal":{"name":"Journal of Industrial Microbiology & Biotechnology","volume":" ","pages":""},"PeriodicalIF":3.2,"publicationDate":"2024-12-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11905756/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143557068","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}