Enzyme and Microbial Technology最新文献

{"title":"Biochemical characterization and structure prediction of the Cerrado soil CRB2(1) metagenomic dioxygenase","authors":"Philippe de Castro Lins ,&nbsp;Pedro Ricardo Vieira Hamann ,&nbsp;Jônatas Cunha Barbosa Lima ,&nbsp;João Alexandre Ribeiro Gonçalves Barbosa ,&nbsp;João Lucas da Silva Correia ,&nbsp;Ikaro Alves de Andrade ,&nbsp;Débora Farage Knupp dos Santos ,&nbsp;Betania Ferraz Quirino ,&nbsp;Ricardo Henrique Krüger","doi":"10.1016/j.enzmictec.2024.110544","DOIUrl":"10.1016/j.enzmictec.2024.110544","url":null,"abstract":"<div><div>Dioxygenases are enzymes involved in the conversion of polyconic aromatic hydroxycarbons (PAHs), attracting significant biotechnological interest for the conversion of recalcitrant organic compounds. Furthermore, few studies show that dioxygenases can take on the function of resistance genes in clones. This enzymatic versatility opens up new opportunities for elucidating the mechanisms of microbial resistance, as well as its biotechnological application. In this work, a <em>Cerrado</em> soil dioxygenase named CRB2(1) was biochemically characterized. The enzyme was shown to have optimal activity at pH 7; a temperature of 30 °C; and using iron ions as a cofactor for substrate cleavage. The kinetic catalytic parameters of CRB2(1) were <em>V</em>_max = 0.02281 µM/min and <em>K</em>_M = 97.6. Its predicted three-dimensional structure obtained using the Modeller software v9.22 based on the crystal structure of gentisate 1,2-dioxygenase from <em>Silicibacter pomeroyi</em> (GDOsp) (PDB ID <span><span>3BU7</span><svg><path></path></svg></span>, resolution 2.80 Å, residues 17–374) revealed substrate binding to the cupin domain, where the active site is located. The analyzed substrates interact directly with the iron ion, coordinated by three histidine residues. Changing the iron ion charge modifies the binding between the active site and the substrates. Currently, there is a demand for enzymes that have biotechnological activities of interest. Metagenomics allows analyzing the biotechnological potential of several organisms at the same time, based on sequence and functional activity analyses.</div></div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"182 ","pages":"Article 110544"},"PeriodicalIF":3.4,"publicationDate":"2024-11-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142617164","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":"Probiotic and functional characterization of newly isolated Lactiplantibacillus plantarum strains from human breast milk and proliferative inhibition potential of metabolites","authors":"Yusuf Alan ,&nbsp;Ali-Osman Keskin ,&nbsp;Mehmet Sönmez","doi":"10.1016/j.enzmictec.2024.110545","DOIUrl":"10.1016/j.enzmictec.2024.110545","url":null,"abstract":"<div><div>Four <em>Lactiplantibacillus plantarum</em> strains newly isolated and identified from human breast milk in Türkiye, have probiotic, functional and proliferative inhibition potential of metabolites against colon cancer cell lines were evaluated. In simulated gastric and intestinal media, all strains exhibited strong probiotic character by showing resistance, although decreasing with time and concentration. The strains were sensitive to penicillin G, rifampin and chloramphenicol and showed antibacterial effect on all pathogenic bacteria. Citric acid, malic acid, tartaric acid, pyruvic acid and fumaric acid were not detected in the strains, while the highest amount of acetic acid was detected. The quantitative-qualitative analysis and structural characterization of exopolysaccharide (EPS) was confirmed and it was determined that the strains synthesized similar amounts. Compared to standard antioxidants, the strains showed less DPPH activity and similar ABTS activity. High amounts of metabolites of the strains showed good antiproliferative effect on Caco-2, while lower amounts showed good antiproliferative effect on the HT-29 cell line. When all the data were considered, it was determined that the strains were close to each other, but the YAAS 23 strain showed slightly better properties. In conclusion, breast milk is a unique environment harboring beneficial bacteria such as <em>L. plantarum</em> for human health.</div></div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"182 ","pages":"Article 110545"},"PeriodicalIF":3.4,"publicationDate":"2024-11-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142638571","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":"Poly-γ-glutamic production by solid-state fermentation of Bacillus natto in ammonia nitrogen movement and soil water retention processes","authors":"Xin Wang ,&nbsp;Jie Gao ,&nbsp;Jiahui Wu ,&nbsp;Xuan Li ,&nbsp;Junxun Li ,&nbsp;Haihong Li ,&nbsp;Songlin Wang","doi":"10.1016/j.enzmictec.2024.110543","DOIUrl":"10.1016/j.enzmictec.2024.110543","url":null,"abstract":"<div><div>A high polyglutamic acid (γ-PGA) producing strain of <em>Bacillus natto UV-40–50</em> was screened by ultraviolet mutagenesis treatment and identified as still belonging to the Bacillus specie. The optimal fermentation medium composition for solid state fermentation (SSF) of <em>B. natto strain UV-40–50 strain</em> was determined by one-way analysis of variance, under which the yield of γ-PGA was 55.19 g/kg, and the presence and molecular weight of γ-PGA in the γ-PGA-purified samples were determined by a series of characterizations. The purification ability of the unseparated solid fermentation product (SFP) on ammonia nitrogen and nitrite in the water column, as well as its effect on soil water retention, germination rate and seedling length of lettuce and cabbage were further investigated. The results showed that the addition of 1 g/m3 SFP could effectively remove more than 60 % of ammonia nitrogen and more than 40 % of nitrite in the water body; the addition of 0.01 % SFP could increase the water retention capacity of cabbage soil by 2.13 times, and increase the water retention capacity of lettuce soil by 12 %; at the same time, the SFP could also significantly increase the germination rate and seedling length of both cabbage and lettuce.</div></div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"182 ","pages":"Article 110543"},"PeriodicalIF":3.4,"publicationDate":"2024-10-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142603884","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":"Advances and prospects for lactic acid production from lignocellulose","authors":"Ruofan Wu,&nbsp;Jiahui Yang,&nbsp;Yujia Jiang,&nbsp;Fengxue Xin","doi":"10.1016/j.enzmictec.2024.110542","DOIUrl":"10.1016/j.enzmictec.2024.110542","url":null,"abstract":"<div><div>Lactic acid is a versatile building block that can be produced via microbial fermentation. Owing to the high optical purity, approximately 90 % of lactic acid is produced by microbes. Recently, the biosynthesis of lactic acid from lignocellulose has concerned much attentions. However, the cost-effective process faces several obstacles because of the complex structure of lignocellulose. This review will comprehensively summarize the state-of-the-art lactic acid production from lignocellulose, including the commonly used lactate-producing microorganisms, the co-utilization of glucose and xylose for the lactic acid production, as well as the lactic acid production from lignocellulose hydrolysate. Furthermore, the strategies regarding the lignocellulosic lactic acid production via consolidated bioprocessing will be also discussed, which can greatly reduce the complexity of the fermentation process.</div></div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"182 ","pages":"Article 110542"},"PeriodicalIF":3.4,"publicationDate":"2024-10-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142567122","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 synthesis of cinnamyl acetate and deacetyl-7-aminocephalosporanic acid by a GDSL-type esterase and its substrate specificity analysis","authors":"Shuqi Xing ,&nbsp;Wei Xie ,&nbsp;Guangli Hu ,&nbsp;Chaocheng Luo ,&nbsp;Hong Zhu ,&nbsp;Laping He ,&nbsp;Cuiqin Li ,&nbsp;Xiao Wang ,&nbsp;Xuefeng Zeng","doi":"10.1016/j.enzmictec.2024.110532","DOIUrl":"10.1016/j.enzmictec.2024.110532","url":null,"abstract":"<div><div>GDSL-type esterases are promising biocatalysts for the food and pharmaceutical industries. Here, a GDSL-type esterase from <em>Aspergillus niger</em> CCTCC No. M2012538 (INANE1) was expressed and purified in <em>Pichia pastoris</em> GS115, and its catalytic performances were evaluated, including the synthesis of cinnamyl acetate and deacetyl-7-aminocephalosporanic acid (D-7-ACA). In addition, molecular docking and molecular dynamics simulations analyzed INANE1's substrate specificity. The substrate specificity profile indicated the recombinant esterase (rINANE1) was an acetylesterase with high specificity for <em>p</em>-nitrophenyl acetate (<em>p</em>-NPA). The rINANE1 exhibited maximum activity at pH 8.0 and 35 °C, where K_m and V_max were calculated as 0.13±0.03 mM and 22.56 ± 0.32 μmoL/min/mg, respectively. The yield of cinnamyl acetate of about 85 % was achieved in 24 h. The conversion rate of 7-aminocephalosporanic acid (7-ACA) could reach 92.71 ± 1.78 % at 25 °C and 2.5 h. Moreover, the INANE1 structure model, molecular docking, and molecular dynamics simulation demonstrated that the pocket of the catalytic triad Ser34, Asn267, and His270 could only accommodate <em>p</em>-NPA. INANE1 may be the first fungi esterase with cinnamyl acetate synthetic activity and 7-ACA hydrolysis activity. Therefore, INANE1 would be a promising enzyme with industrial values.</div></div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"182 ","pages":"Article 110532"},"PeriodicalIF":3.4,"publicationDate":"2024-10-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142544459","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":"Spectrophotometric assay for the screening of selective enzymes towards DHA and EPA ethyl esters hydrolysis","authors":"Hiram Y. Guerrero-Elias ,&nbsp;M. Angeles Camacho-Ruiz ,&nbsp;Ruben Espinosa-Salgado ,&nbsp;Juan Carlos Mateos-Díaz ,&nbsp;Rosa María Camacho-Ruiz ,&nbsp;Ali Asaff-Torres ,&nbsp;Jorge A. Rodríguez","doi":"10.1016/j.enzmictec.2024.110531","DOIUrl":"10.1016/j.enzmictec.2024.110531","url":null,"abstract":"<div><div>Polyunsaturated fatty acids (PUFAs), such as docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA), hold notable significance due to their pharmaceutical relevance. Obtaining PUFAs from diverse sources like vegetables, fish oils, and algae poses challenges due to the mixed fatty acid (FA) composition. Therefore, focusing on particular FAs necessitates purification and resolution processes. To address this, we propose a continuous assay for screening lipases selective for ethyl EPA (E-EPA) or ethyl DHA (E-DHA). Utilizing microplate spectrophotometry, the method enables quantification of liberated fatty acids from ethyl esters (E-EPA or E-DHA). This involves assessing enzyme selectivity by measuring the release of FAs through <em>p</em>-nitrophenolate protonation, either separately for each substrate or in competition with a reference substrate, resorufin acetate. Ten lipases underwent screening, revealing <em>Burkholderia cepacia</em> lipase's (BCL) preference for ethyl DHA hydrolysis (E-EPA/E-DHA = 0.82 ± 0.07 and the lipase selectivity ratio (S) for E-EPA/E-DHA = 0.13 ± 0.04) and <em>Candida antarctica</em> lipase B's (CALB) high specific activity towards both E-EPA and E-DHA (531.14 ± 37.76 and 281.79 ± 2.79 U/mg, respectively) and E-EPA preference (E-EPA/E-DHA = 1.86 ± 0.15 and S E-EPA/E-DHA = 2.59±0.15). <em>Candida rugosa</em> recombinant isoform 4 (rCRLip4) and commercial <em>Candida rugosa</em> lipase (CRL) exhibited significant preference for E-EPA hydrolysis (E-EPA/E-DHA = 2.18 ±0.51 and 2.26 ±0.36, respectively; and S E-EPA/E-DHA = 7.59 ± 1.59 and 7.88 ± 2.13, respectively). Docking analyses of rCRLip4, BCL, and CALB demonstrated no statistically significant differences in activation energies or distances to the catalytic serine; however, they agreed with the experimental results. These findings suggest potential mutagenesis or directed evolution strategies for CALB to enhance E-EPA selectivity, with rCRLip4 emerging as a promising candidate for further investigation. This assay offers a valuable tool for identifying lipases with desired substrate selectivity, with broad implications for pharmaceutical and biotechnological applications.</div></div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"182 ","pages":"Article 110531"},"PeriodicalIF":3.4,"publicationDate":"2024-10-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142561420","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":"New insight into acid-resistant enzymes from natural mutations of Escherichia coli Nissle 1917","authors":"Chengfeng Xue,&nbsp;Wan-Wen Ting,&nbsp;Jiun-Jang Juo,&nbsp;I-Son Ng","doi":"10.1016/j.enzmictec.2024.110526","DOIUrl":"10.1016/j.enzmictec.2024.110526","url":null,"abstract":"<div><div>The probiotic <em>Escherichia coli</em> Nissle 1917 (EcN), known for its superior acid resistance (AR), serves as a promising chassis for live therapeutics due to the effective colonization capabilities. However, the enzymatic activity regarding AR in EcN remains poorly understood. First, we investigated the AR systems of EcN by measuring cell growth under acidic stress and exploring the relationship of mutations to their corresponding enzymatic activities. As a result, the catalytic activity of inducible decarboxylases of GadB, AdiA and CadA, responsible for metabolizing glutamate, arginine, and lysine, exhibited an average 2-fold increase in EcN compared to the reference strain MG1655. Furthermore, we discovered that the glutamate-dependent AR2 system in EcN was meticulously regulated by specific regulons such as GadW. This study not only revealed the physiology of EcN under acidic conditions, but also highlighted that the mutated core enzymes in the AR system of EcN exhibit improved activities.</div></div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"181 ","pages":"Article 110526"},"PeriodicalIF":3.4,"publicationDate":"2024-10-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142497334","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":"On-site cellulase production by Trichoderma reesei RutC-30 to enhance the enzymatic saccharification of ball-milled corn stover","authors":"Yinghui He ,&nbsp;Hui Zhang ,&nbsp;Yeledana Huwati ,&nbsp;Na Shu ,&nbsp;Wei Hu ,&nbsp;Xiwen Jia ,&nbsp;Kaili Ding ,&nbsp;Xueyan Liang ,&nbsp;Luoyang Liu ,&nbsp;Lujia Han ,&nbsp;Weihua Xiao","doi":"10.1016/j.enzmictec.2024.110530","DOIUrl":"10.1016/j.enzmictec.2024.110530","url":null,"abstract":"<div><div>Cellulases are essential for the enzymatic saccharification of lignocellulose. They play a crucial role in breaking down the structure of lignocellulose to obtain fermentable sugars. In this study, we conducted on-site cellulase production by <em>Trichoderma reesei</em> RutC-30 through submerged fermentation. The effects of carbon source, nitrogen source, KH₂PO₄, and mineral elements on cellulase production were evaluated using the hydrolyzed total sugar concentration of ball-milled corn stover as an indicator. The optimal fermentation medium conditions for cellulase production were determined through orthogonal experimental design analysis. Additionally, by optimizing culture conditions, including inoculation, pH, and bottling volume, we achieved a total sugar concentration of 92.25 g/L. After the optimization, the FPA, CMCA, protein, and total sugar concentration increased by 75.49 %, 18.43 %, 89.71 %, and 17.83 %, respectively. Furthermore, corn stover pretreated by different methods was applied to induce cellulase production. Ball-milled and steam-exploded corn stover was identified as suitable incubation carbon sources with total sugar concentration up to 94.31 g/L. Our work exploits the cellulase induced by lignocellulose and then applies it to lignocellulose, enabling the customization and providing a reference for the production of cellulase with corn stover as an inducer.</div></div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"181 ","pages":"Article 110530"},"PeriodicalIF":3.4,"publicationDate":"2024-10-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142497335","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":"Production of bio-indigo from engineered Pseudomonas putida KT2440 harboring tryptophanase and flavin-containing monooxygenase","authors":"Hyun Jin Kim ,&nbsp;Suwon Kim ,&nbsp;Yeda Lee ,&nbsp;Yuni Shin ,&nbsp;Suhye Choi ,&nbsp;Jinok Oh ,&nbsp;Jaeho Jeong ,&nbsp;HyunA Park ,&nbsp;Jungoh Ahn ,&nbsp;Jeong Chan Joo ,&nbsp;Kwon-Young Choi ,&nbsp;Shashi Kant Bhatia ,&nbsp;Yung-Hun Yang","doi":"10.1016/j.enzmictec.2024.110529","DOIUrl":"10.1016/j.enzmictec.2024.110529","url":null,"abstract":"<div><div>Indigo is a unique blue dye that has been used in the textile industry for centuries and is currently mass-produced commercially through chemical synthesis. However, the use of toxic substrates and reducing agents for chemical synthesis is associated with environmental concerns, necessitating the development of eco-friendly alternatives based on microbial production. In this study, a robust industrial strategy for indigo production was developed using <em>Pseudomonas putida</em> KT2440 as the host strain, which is characterized by its excellent ability to degrade aromatic compounds and high resistance to environmental stress. By introducing the genes tryptophanase (<em>tnaA</em>) and Flavin-containing monooxygenase <em>(FMO</em>), a <em>P. putida</em> HI201 strain was constructed to produce indigo from tryptophan. To enhance the indigo yield, culture conditions, including medium composition, temperature, tryptophan concentration, and shaking speed, were optimized. Under optimal conditions such as TB medium, 15 mM tryptophan, 30°C, 200 rpm, <em>P. putida</em> HI201 biosynthesized 1.31 g/L indigo from tryptophan in a fed-batch fermentation system. The introduction of <em>tnaA</em> and <em>FMO</em> genes also enabled the production of indigo in various <em>P. putida</em> species, and the indigo-producing strain had a blue color, which served as a visual indicator. This study presents a strategy for using <em>P. putida</em> as a host for robust and sustainable microbial production of indigo, highlighting the strain's applicability and efficiency in environment friendly dye synthesis.</div></div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"182 ","pages":"Article 110529"},"PeriodicalIF":3.4,"publicationDate":"2024-10-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142497336","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":"Targeted metagenomics – Enrichment for enzymes active on sulfated polysaccharides from seaweeds","authors":"Bjorn Thor Adalsteinsson ,&nbsp;Hörður Guðmundsson ,&nbsp;Andrius Jasilionis ,&nbsp;Morten Schiøtt ,&nbsp;Maria Dalgaard Mikkelsen ,&nbsp;Elísabet Eik Guðmundsdóttir ,&nbsp;Pavithra Sivakumar ,&nbsp;Annika Malmgren ,&nbsp;Tushar Kaushik ,&nbsp;Erik Apelqvist ,&nbsp;Signe Vangsgaard ,&nbsp;Rébecca Leblay ,&nbsp;Ólafur Friðjónsson ,&nbsp;Anne S. Meyer ,&nbsp;Eva Nordberg Karlsson ,&nbsp;Guðmundur Óli Hreggviðsson","doi":"10.1016/j.enzmictec.2024.110528","DOIUrl":"10.1016/j.enzmictec.2024.110528","url":null,"abstract":"<div><div>Seaweeds (macroalgae) are an attractive resource for diverse microbial- and enzymatic production processes. They are abundant, underutilized, cheap, and rich in carbohydrates, and therefore have the potential to be used as a source of mono- or oligosaccharides, and as substrates for industrial fermentation processes. Many seaweed polysaccharides, including the sulfated polysaccharides ulvan and fucoidan, are however complex and heterogenous in structure, and there are currently few enzymes available to modify them, and understanding of their enzymatic depolymerization remains limited. The present study aimed to identify and characterize robust fucoidanases and ulvan lyases. Metagenomes were obtained from microbial enrichments from an intertidal hot-spring, genes identified that expressed putative fucoidanases and ulvan lyases, and following gene cloning and expression, the respective enzymes were screened for enzymatic activity. Consistent with their origin, the identified protein sequences were considerably divergent from previously characterized enzymes, with a 44 % average maximal sequence identity. In total, the study resulted in the characterization of 10 new fucoidanases (GH107 and GH168 families) and 8 new ulvan lyases (PL24, PL25 and PL40 families). Notably, the new fucoidanases appeared to have functional specificity towards fucoidan containing α-1,3 linked L-fucosyl and several functioned at high temperature. The study contributes a metagenomics-based approach to identify new seaweed polysaccharide degrading enzymes and an increased understanding of the diversity of such enzymes, which may have implications for the realization of biotechnology based valorization of seaweed biomass.</div></div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"182 ","pages":"Article 110528"},"PeriodicalIF":3.4,"publicationDate":"2024-10-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142578510","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}