@article{Roman2024,

title = {Plastic-degrading microbial communities reveal novel microorganisms, pathways, and biocatalysts for polymer degradation and bioplastic production},

author = {Ellen Karen Barreto Roman and Murilo Antonio Ramos and Geizecler Tomazetto and Bruno Botega Foltran and Matheus Henrique Galvão and Iara Ciancaglini and Robson Tramontina and Felipe de Almeida Rodrigues and Larissa Soares da Silva and Ana Luiza Hernandes Sandano and Diógenes G. da S. Fernandes and Dnane Vieira Almeida and Denicezar Angelo Baldo and José Martins de Oliveira Junior and Wanius Garcia and André Damasio and Fabio Marcio Squina},

url = {https://www.sciencedirect.com/science/article/abs/pii/S0048969724050253},

doi = {10.1016/j.scitotenv.2024.174876},

issn = {0048-9697},

year  = {2024},

date = {2024-11-00},

urldate = {2024-11-00},

journal = {Science of The Total Environment},

volume = {949},

publisher = {Elsevier BV},

abstract = {Plastics derived from fossil fuels are used ubiquitously owing to their exceptional physicochemical characteristics. However, the extensive and short-term use of plastics has caused environmental challenges. The biotechnological plastic conversion can help address the challenges related to plastic pollution, offering sustainable alternatives that can operate using bioeconomic concepts and promote socioeconomic benefits. In this context, using soil from a plastic-contaminated landfill, two consortia were established (ConsPlastic-A and -B) displaying versatility in developing and consuming polyethylene or polyethylene terephthalate as the carbon source of nutrition. The ConsPlastic-A and -B metagenomic sequencing, taxonomic profiling, and the reconstruction of 79 draft bacterial genomes significantly expanded the knowledge of plastic-degrading microorganisms and enzymes, disclosing novel taxonomic groups associated with polymer degradation. The microbial consortium was utilized to obtain a novel Pseudomonas putida strain (BR4), presenting a striking metabolic arsenal for aromatic compound degradation and assimilation, confirmed by genomic analyses. The BR4 displays the inherent capacity to degrade polyethylene terephthalate (PET) and produce polyhydroxybutyrate (PHB) containing hydroxyvalerate (HV) units that contribute to enhanced copolymer properties, such as increased flexibility and resistance to breakage, compared with pure PHB. Therefore, BR4 is a promising strain for developing a bioconsolidated plastic depolymerization and upcycling process. Collectively, our study provides insights that may extend beyond the artificial ecosystems established during our experiments and supports future strategies for effectively decomposing and valorizing plastic waste. Furthermore, the functional genomic analysis described herein serves as a valuable guide for elucidating the genetic potential of microbial communities and microorganisms in plastic deconstruction and upcycling.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{Almeida2024,

title = {Unveiling the crystal structure of thermostable dienelactone hydrolase exhibiting activity on terephthalate esters},

author = {Dnane Vieira Almeida and Iara Ciancaglini and Ana Luiza Hernandes Sandano and Ellen K.B. Roman and Viviane Brito Andrade and Ana Bárbara Nunes and Robson Tramontina and Viviam Moura da Silva and Frank Gabel and Thamy L.R. Corrêa and André Damasio and João Renato Carvalho Muniz and Fabio Marcio Squina and Wanius Garcia},

url = {https://www.sciencedirect.com/science/article/abs/pii/S0141022924001054},

doi = {10.1016/j.enzmictec.2024.110498},

issn = {0141-0229},

year  = {2024},

date = {2024-10-00},

urldate = {2024-10-00},

journal = {Enzyme and Microbial Technology},

volume = {180},

publisher = {Elsevier BV},

abstract = {Dienelactone hydrolase (DLH) is one of numerous hydrolytic enzymes with an α/β-hydrolase fold, which catalyze the hydrolysis of dienelactone to maleylacetate. The DLHs share remarkably similar tertiary structures and a conserved arrangement of catalytic residues. This study presents the crystal structure and comprehensive functional characterization of a novel thermostable DLH from the bacterium Hydrogenobacter thermophilus (HtDLH). The crystal structure of the HtDLH, solved at a resolution of about 1.67 Å, exhibits a canonical α/β-hydrolase fold formed by eight β-sheet strands in the core, with one buried α-helix and six others exposed to the solvent. The structure also confirmed the conserved catalytic triad of DHLs formed by Cys121, Asp170, and His202 residues. The HtDLH forms stable homodimers in solution. Functional studies showed that HtDLH has the expected esterase activity over esters with short carbon chains, such as p-nitrophenyl acetate, reaching optimal activity at pH 7.5 and 70 °C. Furthermore, HtDLH maintains more than 50 % of its activity even after incubation at 90 °C for 16 h. Interestingly, HtDLH exhibits catalytic activity towards polyethylene terephthalate (PET) monomers, including bis-1,2-hydroxyethyl terephthalate (BHET) and 1-(2-hydroxyethyl) 4-methyl terephthalate, as well as other aliphatic and aromatic esters. These findings associated with the lack of activity on amorphous PET indicate that HtDLH has characteristic of a BHET-degrading enzyme. This work expands our understanding of enzyme families involved in PET degradation, providing novel insights for plastic biorecycling through protein engineering, which could lead to eco-friendly solutions to reduce the accumulation of plastic in landfills and natural environments.



},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{Silva2024b,

title = {Fabrication and characterization of physically crosslinked alginate/chitosan-based hydrogel loaded with neomycin for the treatment of skin infections caused by Staphylococcus aureus},

author = {Lucas dos Santos Silva and Beatriz Gomes Vila Nova and Carlos Eduardo Morais de Sousa and Raphael Guedes Silva and Layse Ribeiro de Sousa Carvalho and Izadora Souza Soeiro Silva and Pedro Henrique de Aguiar Moreira and Andres Felipe Millan Cardenas and Cristina de Andrade Monteiro and Aryane Tofanello and Wanius Garcia and Claudener Souza Teixeira and Luís Cláudio Nascimento da Silva},

url = {https://www.sciencedirect.com/science/article/abs/pii/S0141813024033828},

doi = {10.1016/j.ijbiomac.2024.132577},

issn = {0141-8130},

year  = {2024},

date = {2024-06-00},

urldate = {2024-06-00},

journal = {International Journal of Biological Macromolecules},

volume = {271},

publisher = {Elsevier BV},

abstract = {Staphylococcus aureus is a pathogen widely involved in wound infection due to its ability to release several virulence factors that impair the skin healing process, as well as its mechanism of drug resistance. Herein, sodium alginate and chitosan were combined to produce a hydrogel for topical delivery of neomycin to combat S. aureus associated with skin complications. The hydrogel was formulated by combining sodium alginate (50 mg/mL) and chitosan (50 mg/mL) solutions in a ratio of 9:1 (HBase). Neomycin was added to HBase to achieve a concentration of 0.4 mg/mL (HNeo). The incorporation of neomycin into the product was confirmed by scanning electron microscopy, FTIR and TGA analysis. The hydrogels produced are homogeneous, have a high swelling capacity, and show biocompatibility using erythrocytes and fibroblasts as models. The formulations showed physicochemical and pharmacological stability for 60 days at 4 ± 2 °C. HNeo totally inhibited the growth of S. aureus after 4 h. The antimicrobial effects were confirmed using ex vivo (porcine skin) and in vivo (murine) wound infection models. Furthermore, the HNeo-treated mice showed lower severity scores than those treated with HBase. Taken together, the obtained results present a new low-cost bioproduct with promising applications in treating infected wounds.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{Andrade2024,

title = {Enzymatic and biophysical characterization of a novel modular cellulosomal GH5 endoglucanase multifunctional from the anaerobic gut fungus Piromyces finnis},

author = {Viviane Brito Andrade and Geizecler Tomazetto and Dnane Vieira Almeida and Robson Tramontina and Fabio Marcio Squina and Wanius Garcia},

url = {https://www.sciencedirect.com/science/article/abs/pii/S1570963923000778},

doi = {10.1016/j.bbapap.2023.140963},

issn = {1570-9639},

year  = {2024},

date = {2024-01-00},

urldate = {2024-01-00},

journal = {Biochimica et Biophysica Acta (BBA) - Proteins and Proteomics},

volume = {1872},

number = {1},

publisher = {Elsevier BV},

abstract = {Cellulases from anaerobic fungi are enzymes less-studied biochemically and structurally than cellulases from bacteria and aerobic fungi. Currently, only thirteen GH5 cellulases from anaerobic fungi were biochemically characterized and two crystal structures were reported. In this context, here, we report the functional and biophysical characterization of a novel multi-modular cellulosomal GH5 endoglucanase from the anaerobic gut fungus Piromyces finnis (named here PfGH5). Multiple sequences alignments indicate that PfGH5 is composed of a GH5 catalytic domain and a CBM1 carbohydrate-binding module connected through a CBM10 dockerin module. Our results showed that PfGH5 is an endoglucanase from anaerobic fungus with a large spectrum of activity. PfGH5 exhibited preference for hydrolysis of oat β-glucan, followed by galactomannan, carboxymethyl cellulose, mannan, lichenan and barley β-glucan, therefore displaying multi-functionality. For oat β-glucan, PfGH5 reaches its optimum enzymatic activity at 40 °C and pH 5.5, with Km of 7.1 μM. Ion exchange chromatography analyzes revealed the production of oligosaccharides with a wide degree of polymerization indicated that PfGH5 has endoglucanase activity. The ability to bind and cleave different types of carbohydrates evidence the potential of PfGH5 for use in biotechnology and provide a useful basis for future investigation and application of new anaerobic fungi enzymes.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{Bazán2023,

title = {Development and characterization of alginate-derived bioadhesive films incorporated with anti-infective lectins for application in the treatment of oral candidiasis},

author = {José Manuel Noguera Bazán and Vitor Lopes Chagas and Raphael Guedes Silva and Izadora Souza Soeiro Silva and João Guilherme Nantes Araujo and Lucas dos Santos Silva and Karla Lilian Rodrigues Batista and Romério Rodrigues dos Santos Silva and Maria Tereza dos Santos Correia and Joicy Cortez de Sá Sousa and Cristina de Andrade Monteiro and Aryane Tofanello and Wanius Garcia and Edilausson Moreno Carvalho and Claudener Souza Teixeira and Luís Cláudio Nascimento da Silva},

url = {https://www.sciencedirect.com/science/article/abs/pii/S1773224723009668},

doi = {10.1016/j.jddst.2023.105114},

issn = {1773-2247},

year  = {2023},

date = {2023-12-00},

urldate = {2023-12-00},

journal = {Journal of Drug Delivery Science and Technology},

volume = {90},

publisher = {Elsevier BV},

abstract = {This study reports the characterization of alginate-derived bioadhesive films containing the lectins ConBr (from Canavalia brasiliensis Mart.) and MaL (from Machaerium acutifolium Vogel.). The anti-infective properties were evaluated using Tenebrio molitor larvae infected with an oral isolate of Candida albicans (CA40) (1 × 104 yeast/animal). Untreated larvae showed a median survival of 1.5 days and a survival rate of 40%. Both lectins (128 μg/kg/larvae) showed prophylactic effects (survival rates: 80%–90%); however, only MaL significantly increased larval survival (70%) in the treatment assays. Bioadhesive films were developed using sodium alginate, and each lectin was incorporated at 50 μg/mL of filmogenic solution. All films (pH 6.4) showed similar micromorphology with a slightly granular appearance. The lectins-loaded films exhibited more irregular surfaces than the control. The proteins were released in similar patterns. FTIR analysis confirmed the modification of sodium alginate due to the chemical interaction with lectins. All films exhibited a two-step weight loss pattern in TGA analysis. In the ex vivo mucoadhesive assay using bovine jugal mucosa, the highest value of maximum detachment force was found for ConBr-loaded films (0.56 ± 0.04 N) (p < 0.01). The other formulations showed similar adhesiveness (0.48 and 0.47 N). In general, the lectins did not induce significant changes in the physicochemical and mechanical characteristics of the alginate films. The data obtained in this work show, for the first time, the in vivo anti-infective effects of ConBr and MaL against C. albicans. The lectins-containing alginate films presented parameters compatible with their application in the oral cavity.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{dosSantos2022,

title = {Recombinant expression, purification and characterization of an active bacterial feruloyl-CoA synthase with potential for application in vanillin production},

author = {Otávio Augusto Leitão dos Santos and Thiago Augusto Gonçalves and Victoria Sodré and Nathalia Vilela and Geizecler Tomazetto and Fabio M. Squina and Wanius Garcia},

url = {https://www.sciencedirect.com/science/article/pii/S1046592822000663},

doi = {10.1016/j.pep.2022.106109},

issn = {1046-5928},

year  = {2022},

date = {2022-09-00},

urldate = {2022-09-00},

journal = {Protein Expression and Purification},

volume = {197},

publisher = {Elsevier BV},

abstract = {The ferulic acid (FA) represents a high-value molecule with applications in the cosmetic and pharmaceutical industries. This aromatic molecule is derived from lignin and can be enzymatically converted in other commercially interesting molecules, such as vanillin and bioplastics. This process starts with a common step of FA activation via CoA-thioesterification, catalyzed by feruloyl-CoA synthetases. Therefore, here, we report the successfully expression, purification as well as the initial structural and biochemical characterization of a stable, correctly folded, and catalytically active bacterial feruloyl-CoA synthase (here named FCS3) isolated from a lignin-degrading microbial consortium. The purification of recombinant FCS3 to near homogeneity was achieved using affinity chromatography. The FCS3 structure is composed of a mixture of α and β secondary structures and most likely forms stable homodimers in solution. The FCS3 presented a notable structural stability at alkaline pH values and it was able to convert FA and coenzyme A (CoA) into feruloyl-CoA complex at room temperature. This study should provide a useful basis for future biotechnological applications of FCS3, especially in the field of conversion of lignin-derived FA into high value compounds.



    Previous article in issue},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{dosSantos2022b,

title = {Recombinant expression, purification and characterization of an active bacterial feruloyl-CoA synthase with potential for application in vanillin production},

author = {Otávio Augusto Leitão dos Santos and Thiago Augusto Gonçalves and Victoria Sodré and Nathalia Vilela and Geizecler Tomazetto and Fabio M. Squina and Wanius Garcia},

url = {https://www.sciencedirect.com/science/article/abs/pii/S1046592822000663},

doi = {10.1016/j.pep.2022.106109},

issn = {1046-5928},

year  = {2022},

date = {2022-09-00},

urldate = {2022-09-00},

journal = {Protein Expression and Purification},

volume = {197},

publisher = {Elsevier BV},

abstract = {The ferulic acid (FA) represents a high-value molecule with applications in the cosmetic and pharmaceutical industries. This aromatic molecule is derived from lignin and can be enzymatically converted in other commercially interesting molecules, such as vanillin and bioplastics. This process starts with a common step of FA activation via CoA-thioesterification, catalyzed by feruloyl-CoA synthetases. Therefore, here, we report the successfully expression, purification as well as the initial structural and biochemical characterization of a stable, correctly folded, and catalytically active bacterial feruloyl-CoA synthase (here named FCS3) isolated from a lignin-degrading microbial consortium. The purification of recombinant FCS3 to near homogeneity was achieved using affinity chromatography. The FCS3 structure is composed of a mixture of α and β secondary structures and most likely forms stable homodimers in solution. The FCS3 presented a notable structural stability at alkaline pH values and it was able to convert FA and coenzyme A (CoA) into feruloyl-CoA complex at room temperature. This study should provide a useful basis for future biotechnological applications of FCS3, especially in the field of conversion of lignin-derived FA into high value compounds.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{Liberato2022,

title = {Unique properties of a Dictyostelium discoideum carbohydrate-binding module expand our understanding of CBM–ligand interactions},

author = {Marcelo Vizona Liberato and Bruna Medeia Campos and Geizecler Tomazetto and Lucy Isobel Crouch and Wanius Garcia and Ana Carolina de Mattos Zeri and David Nichol Bolam and Fabio Marcio Squina},

url = {https://www.jbc.org/article/S0021-9258(22)00331-3/fulltext},

doi = {10.1016/j.jbc.2022.101891},

issn = {0021-9258},

year  = {2022},

date = {2022-05-00},

urldate = {2022-05-00},

journal = {Journal of Biological Chemistry},

volume = {298},

number = {5},

publisher = {Elsevier BV},

abstract = {Abstract

Deciphering how enzymes interact, modify, and recognize carbohydrates has long been a topic of interest in academic, pharmaceutical, and industrial research. Carbohydrate-binding modules (CBMs) are noncatalytic globular protein domains attached to carbohydrate-active enzymes that strengthen enzyme affinity to substrates and increase enzymatic efficiency via targeting and proximity effects. CBMs are considered auspicious for various biotechnological purposes in textile, food, and feed industries, representing valuable tools in basic science research and biomedicine. Here, we present the first crystallographic structure of a CBM8 family member (CBM8), DdCBM8, from the slime mold Dictyostelium discoideum, which was identified attached to an endo-β-1,4-glucanase (glycoside hydrolase family 9). We show that the planar carbohydrate-binding site of DdCBM8, composed of aromatic residues, is similar to type A CBMs that are specific for crystalline (multichain) polysaccharides. Accordingly, pull-down assays indicated that DdCBM8 was able to bind insoluble forms of cellulose. However, affinity gel electrophoresis demonstrated that DdCBM8 also bound to soluble (single chain) polysaccharides, especially glucomannan, similar to type B CBMs, although it had no apparent affinity for oligosaccharides. Therefore, the structural characteristics and broad specificity of DdCBM8 represent exceptions to the canonical CBM classification. In addition, mutational analysis identified specific amino acid residues involved in ligand recognition, which are conserved throughout the CBM8 family. This advancement in the structural and functional characterization of CBMs contributes to our understanding of carbohydrate-active enzymes and protein–carbohydrate interactions, pushing forward protein engineering strategies and enhancing the potential biotechnological applications of glycoside hydrolase accessory modules.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{Liberato2022b,

title = {Unique properties of a Dictyostelium discoideum carbohydrate-binding module expand our understanding of CBM–ligand interactions},

author = {Marcelo Vizona Liberato and Bruna Medeia Campos and Geizecler Tomazetto and Lucy Isobel Crouch and Wanius Garcia and Ana Carolina de Mattos Zeri and David Nichol Bolam and Fabio Marcio Squina},

url = {https://www.jbc.org/article/S0021-9258(22)00331-3/fulltext},

doi = {10.1016/j.jbc.2022.101891},

issn = {0021-9258},

year  = {2022},

date = {2022-05-00},

urldate = {2022-05-00},

journal = {Journal of Biological Chemistry},

volume = {298},

number = {5},

publisher = {Elsevier BV},

abstract = {Deciphering how enzymes interact, modify, and recognize carbohydrates has long been a topic of interest in academic, pharmaceutical, and industrial research. Carbohydrate-binding modules (CBMs) are noncatalytic globular protein domains attached to carbohydrate-active enzymes that strengthen enzyme affinity to substrates and increase enzymatic efficiency via targeting and proximity effects. CBMs are considered auspicious for various biotechnological purposes in textile, food, and feed industries, representing valuable tools in basic science research and biomedicine. Here, we present the first crystallographic structure of a CBM8 family member (CBM8), DdCBM8, from the slime mold Dictyostelium discoideum, which was identified attached to an endo-β-1,4-glucanase (glycoside hydrolase family 9). We show that the planar carbohydrate-binding site of DdCBM8, composed of aromatic residues, is similar to type A CBMs that are specific for crystalline (multichain) polysaccharides. Accordingly, pull-down assays indicated that DdCBM8 was able to bind insoluble forms of cellulose. However, affinity gel electrophoresis demonstrated that DdCBM8 also bound to soluble (single chain) polysaccharides, especially glucomannan, similar to type B CBMs, although it had no apparent affinity for oligosaccharides. Therefore, the structural characteristics and broad specificity of DdCBM8 represent exceptions to the canonical CBM classification. In addition, mutational analysis identified specific amino acid residues involved in ligand recognition, which are conserved throughout the CBM8 family. This advancement in the structural and functional characterization of CBMs contributes to our understanding of carbohydrate-active enzymes and protein–carbohydrate interactions, pushing forward protein engineering strategies and enhancing the potential biotechnological applications of glycoside hydrolase accessory modules.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{Gonçalves2022,

title = {Applying biochemical and structural characterization of hydroxycinnamate catabolic enzymes from soil metagenome for lignin valorization strategies},

author = {Thiago Augusto Gonçalves and Victoria Sodré and Stephanie Nemesio da Silva and Nathalia Vilela and Geizecler Tomazetto and Juscemácia Nascimento Araujo and João Renato C. Muniz and Taícia Pacheco Fill and André Damasio and Wanius Garcia and Fabio Marcio Squina},

url = {https://link.springer.com/article/10.1007/s00253-022-11885-3},

doi = {10.1007/s00253-022-11885-3},

issn = {1432-0614},

year  = {2022},

date = {2022-04-00},

urldate = {2022-04-00},

journal = {Appl Microbiol Biotechnol},

volume = {106},

number = {7},

pages = {2503--2516},

publisher = {Springer Science and Business Media LLC},

abstract = {The biocatalytic production of fuels and chemicals from plant biomass represents an attractive alternative to fossil fuel-based refineries. In this context, the mining and characterization of novel biocatalysts can promote disruptive innovation opportunities in the field of lignocellulose conversion and valorization. In the present work, we conducted the biochemical and structural characterization of two novel hydroxycinnamic acid catabolic enzymes, isolated from a lignin-degrading microbial consortium, a feruloyl-CoA synthetase, and a feruloyl-CoA hydratase-lyase, named LM-FCS2 and LM-FCHL2, respectively. Besides establishing the homology model structures for novel FCS and FCHL members with unique characteristics, the enzymes presented interesting biochemical features: LM-FCS2 showed stability in alkaline pHs and was able to convert a wide array of p-hydroxycinnamic acids to their respective CoA-thioesters, including sinapic acid; LM-FCHL2 efficiently converted feruloyl-CoA and p-coumaroyl-CoA into vanillin and 4-hydroxybenzaldehyde, respectively, and could produce vanillin directly from ferulic acid. The coupled reaction of LM-FCS2 and LM-FCHL2 produced vanillin, not only from commercial ferulic acid but also from a crude lignocellulosic hydrolysate. Collectively, this work illuminates the structure and function of two critical enzymes involved in converting ferulic acid into high-value molecules, thus providing valuable concepts applied to the development of plant biomass biorefineries.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{Gonçalves2022b,

title = {Applying biochemical and structural characterization of hydroxycinnamate catabolic enzymes from soil metagenome for lignin valorization strategies},

author = {Thiago Augusto Gonçalves and Victoria Sodré and Stephanie Nemesio da Silva and Nathalia Vilela and Geizecler Tomazetto and Juscemácia Nascimento Araujo and João Renato C. Muniz and Taícia Pacheco Fill and André Damasio and Wanius Garcia and Fabio Marcio Squina},

url = {https://link.springer.com/article/10.1007/s00253-022-11885-3},

doi = {10.1007/s00253-022-11885-3},

issn = {1432-0614},

year  = {2022},

date = {2022-04-00},

urldate = {2022-04-00},

journal = {Appl Microbiol Biotechnol},

volume = {106},

number = {7},

pages = {2503--2516},

publisher = {Springer Science and Business Media LLC},

abstract = {The biocatalytic production of fuels and chemicals from plant biomass represents an attractive alternative to fossil fuel-based refineries. In this context, the mining and characterization of novel biocatalysts can promote disruptive innovation opportunities in the field of lignocellulose conversion and valorization. In the present work, we conducted the biochemical and structural characterization of two novel hydroxycinnamic acid catabolic enzymes, isolated from a lignin-degrading microbial consortium, a feruloyl-CoA synthetase, and a feruloyl-CoA hydratase-lyase, named LM-FCS2 and LM-FCHL2, respectively. Besides establishing the homology model structures for novel FCS and FCHL members with unique characteristics, the enzymes presented interesting biochemical features: LM-FCS2 showed stability in alkaline pHs and was able to convert a wide array of p-hydroxycinnamic acids to their respective CoA-thioesters, including sinapic acid; LM-FCHL2 efficiently converted feruloyl-CoA and p-coumaroyl-CoA into vanillin and 4-hydroxybenzaldehyde, respectively, and could produce vanillin directly from ferulic acid. The coupled reaction of LM-FCS2 and LM-FCHL2 produced vanillin, not only from commercial ferulic acid but also from a crude lignocellulosic hydrolysate. Collectively, this work illuminates the structure and function of two critical enzymes involved in converting ferulic acid into high-value molecules, thus providing valuable concepts applied to the development of plant biomass biorefineries.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{daS.Fernandes2021,

title = {Cytotoxicity and Antimicrobial Properties of Photosynthesized Silver Chloride Nanoparticles Using Plant Extract from Stryphnodendron adstringens (Martius) Coville},

author = {Diógenes G. da S. Fernandes and Viviane Brito Andrade and Letícia Neves Lucena and Felipe Nogueira Ambrosio and André L. M. de Souza and Bruno L. Batista and Wallace Rosado Rolim and Amedea Barozzi Seabra and Christiane B. Lombello and Fernanda Dias da Silva and Wanius Garcia},

url = {https://link.springer.com/article/10.1007/s10876-021-02011-w},

doi = {10.1007/s10876-021-02011-w},

issn = {1572-8862},

year  = {2022},

date = {2022-03-00},

urldate = {2022-03-00},

journal = {J Clust Sci},

volume = {33},

number = {2},

pages = {687--695},

publisher = {Springer Science and Business Media LLC},

abstract = {Stryphnodendron adstringens (Martius) Coville is a medicinal plant described as having pharmacological properties as anti-inflammatory and antimicrobial activities. Silver chloride nanoparticles (AgCl-NPs) have shown great potential for biomedical applications with efficient antimicrobial properties. Here, we report the photosynthesis of AgCl-NPs using plant extract from S. adstringens (SaAgCl-NPs) and their cytotoxic and antimicrobial activities. We photosynthesized SaAgCl-NPs nearly spherical with low heterogeneity in size and enveloped by an organic material layer responsible for colloidal stability. SaAgCl-NPs was non-cytotoxic against mammalian VERO cells; however, SaAgCl-NPs presented remarkable antifungal activity against the pathogenic yeast Cryptococcus neoformans (MIC80 of 0.32 µg/mL) the causative agent of human cryptococcosis. Notable antibacterial activity was observed against Gram-negative bacteria Pseudomonas aeruginosa (MIC80 of 2.56 µg/mL) e Serratia marcescens (MIC80 of 20.48 µg/mL) both microorganisms associated with a variety of human infections, in particular pneumonia. In contrast, Gram-positive bacteria Staphylococcus aureus and Staphylococcus epidermidis, microorganisms that cause pathologies as skin infections, were less susceptible to the SaAgCl-NPs both with MIC80 of 40.93 µg/mL. Thus, SaAgCl-NPs represents an organic–inorganic hybrid nanomaterial with very low cytotoxicity against mammalian cells and high antimicrobial efficiency against pathogenic microorganisms and may be explored as an alternative to antimicrobial drugs.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{FrancoCairo2022,

title = {The periplasmic expression and purification of AA15 lytic polysaccharide monooxygenases from insect species in Escherichia coli},

author = {João Paulo L. Franco Cairo and Dnane V. Almeida and André Damasio and Wanius Garcia and Fabio M. Squina},

url = {https://www.sciencedirect.com/science/article/pii/S1046592821001777},

doi = {10.1016/j.pep.2021.105994},

issn = {1046-5928},

year  = {2022},

date = {2022-02-00},

urldate = {2022-02-00},

journal = {Protein Expression and Purification},

volume = {190},

publisher = {Elsevier BV},

abstract = {Lytic polysaccharide monooxygenases (LPMOs) are metalloenzymes that cleave structural polysaccharides through an oxidative mechanism. The enzymatic activity of LPMOs relies on the presence of a Cu2+ histidine-brace motif in their flat catalytic surface. Upon reduction by an external electron donor and in the presence of its co-substrates, O2 or H2O2, LPMOs can generate reactive oxygen species to oxidize the substrates. Fungal and bacterial LPMOs are involved in the catabolism of polysaccharides, such as chitin, cellulose, and hemicelluloses, and virulence mechanisms. Based on the reports on the discovery of LPMOs from the family AA15 in termites, firebrats, and flies, the functional role of the LPMO in the biosphere could expand, as these enzymes may be correlated with chitin remodeling and molting in insects. However, there is limited knowledge of AA15 LPMOs due to difficulties in recombinant expression of soluble proteins and purification protocols. In this study, we describe a protocol for the cloning, expression, and purification of insect AA15 LPMOs from Arthropoda, mainly from termites, followed by the expression and purification of an AA15 LPMO from the silkworm Bombyx mori, which contains a relatively high number of disulfide bonds. We also report the recombinant expression and purification of a protein with homology to AA15 family from the western European honeybee Apis mellifera, an LPMO-like enzyme lacking the canonical histidine brace. Therefore, this work can support future studies concerning the role of LPMOs in the biology of insects and inspire molecular entomologists and insect biochemists in conducting activities in this field.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{FrancoCairo2022b,

title = {The periplasmic expression and purification of AA15 lytic polysaccharide monooxygenases from insect species in Escherichia coli},

author = {João Paulo L. Franco Cairo and Dnane V. Almeida and André Damasio and Wanius Garcia and Fabio M. Squina},

doi = {10.1016/j.pep.2021.105994},

issn = {1046-5928},

year  = {2022},

date = {2022-02-00},

urldate = {2022-02-00},

journal = {Protein Expression and Purification},

volume = {190},

publisher = {Elsevier BV},

abstract = {Lytic polysaccharide monooxygenases (LPMOs) are metalloenzymes that cleave structural polysaccharides through an oxidative mechanism. The enzymatic activity of LPMOs relies on the presence of a Cu2+ histidine-brace motif in their flat catalytic surface. Upon reduction by an external electron donor and in the presence of its co-substrates, O2 or H2O2, LPMOs can generate reactive oxygen species to oxidize the substrates. Fungal and bacterial LPMOs are involved in the catabolism of polysaccharides, such as chitin, cellulose, and hemicelluloses, and virulence mechanisms. Based on the reports on the discovery of LPMOs from the family AA15 in termites, firebrats, and flies, the functional role of the LPMO in the biosphere could expand, as these enzymes may be correlated with chitin remodeling and molting in insects. However, there is limited knowledge of AA15 LPMOs due to difficulties in recombinant expression of soluble proteins and purification protocols. In this study, we describe a protocol for the cloning, expression, and purification of insect AA15 LPMOs from Arthropoda, mainly from termites, followed by the expression and purification of an AA15 LPMO from the silkworm Bombyx mori, which contains a relatively high number of disulfide bonds. We also report the recombinant expression and purification of a protein with homology to AA15 family from the western European honeybee Apis mellifera, an LPMO-like enzyme lacking the canonical histidine brace. Therefore, this work can support future studies concerning the role of LPMOs in the biology of insects and inspire molecular entomologists and insect biochemists in conducting activities in this field.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{https://doi.org/10.1002/bab.2416,

title = {Photosynthesis of hybrid silver-based nanoparticles mediated lectins and evaluation of their hemagglutinating properties},

author = {Diógenes G. S Fernandes and Graziella S. Capo and Aryane Tofanello and Carlos E. Castro and Bruno B. Foltran and Fabio M. Squina and Maria H. C Santos and Romério R. S. Silva and Claudener S. Teixeira and Wanius Garcia},

url = {https://iubmb.onlinelibrary.wiley.com/doi/abs/10.1002/bab.2416},

doi = {https://doi.org/10.1002/bab.2416},

year  = {2022},

date = {2022-01-01},

urldate = {2023-01-01},

journal = {Biotechnology and Applied Biochemistry},

volume = {70},

number = {3},

pages = {1015-1023},

abstract = {Abstract Lectins are carbohydrate-binding proteins belonging to the Leguminosae family. In this family stand out proteins extracted from species belonging to Diocleinae subtribe, which includes, for example, the seed lectin from Dioclea violacea (DVL) and the jack bean lectin Concanavalin A (ConA). Here, we report the photosynthesis of silver/silver chloride nanoparticles (NPs) assisted by ConA and DVL. The syntheses were simple processes using a green-chemistry approach. Under electron microscopy, NPs heterogeneous in size, nearly spherical and covered by a thin lectin corona, were observed. Both NPs assisted by lectins were capable to cause strong rabbit erythrocytes agglutination with the same titers of hemagglutinating activities. These results indicate that both lectins maintained their biological activities even after association with the NPs and therefore are able to interact with biological membrane carbohydrates. However, for rabbit erythrocytes treated with proteolytic enzymes were observed different titers of hemagglutinating activities, suggesting differences in the spatial arrangement of the lectins on the surface of the NPs. This study provides evidences that these hybrid lectin-coated silver/silver chloride NPs can be used for selective recognition and interaction with membrane carbohydrates and others biotechnological applications.},

keywords = {carbohydrates recognition, halide nanoparticles, hemagglutinating activity, photosynthesis, plant lectins},

pubstate = {published},

tppubtype = {article}

}

@article{deLima2021c,

title = {Crystal Structure of a Sucrose-6-phosphate Hydrolase from Lactobacillus gasseri with Potential Applications in Fructan Production and the Food Industry},

author = {Mariana Z. T. de Lima and Leonardo R. de Almeida and Alain M. Mera and Amanda Bernardes and Wanius Garcia and João R. C. Muniz},

url = {https://pubs.acs.org/doi/full/10.1021/acs.jafc.1c03901},

doi = {10.1021/acs.jafc.1c03901},

issn = {1520-5118},

year  = {2021},

date = {2021-09-08},

urldate = {2021-09-08},

journal = {J. Agric. Food Chem.},

volume = {69},

number = {35},

pages = {10223--10234},

publisher = {American Chemical Society (ACS)},

abstract = {Fructooligosaccharides (FOSs) are polymers of fructose with a prebiotic activity because of their production and fermentation by bacteria that inhabit the gastrointestinal tract and are widely used in the industry and new functional foods. Lactobacillus gasseri stands out as an important homofermentative microorganism related to FOS production, and its potential applications in the industry are undeniable. In this study, we report the production and characterization of a sucrose-6-phosphate hydrolase from L. gasseri belonging to the GH32 family. Apo-LgAs32 and LgAs32 complexed with β-d-fructose structures were determined at a resolution of 1.94 and 1.84 Å, respectively. The production of FOS, fructans, 1-kestose, and nystose by the recombinant LgAs32, using sucrose as a substrate, shown in this study is very promising. When compared to its homologous enzyme from Lactobacillus reuteri, the production of 1-kestose by LgAs32 is increased; thus, LgAs32 can be considered as an alternative in fructan production and other industrial applications.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{Santos2021d,

title = {Enhanced antibacterial activity of the gentamicin against multidrug-resistant strains when complexed with Canavalia ensiformis lectin},

author = {Valdenice F. Santos and Ana C.J. Araújo and Priscilla R. Freitas and Ana L.P. Silva and Ana L.E. Santos and Bruno Anderson Matias da Rocha and Romério R.S. Silva and Dnane V. Almeida and Wanius Garcia and Henrique D.M. Coutinho and Claudener S. Teixeira},

url = {https://www.sciencedirect.com/science/article/pii/S0882401020310056},

doi = {10.1016/j.micpath.2020.104639},

issn = {0882-4010},

year  = {2021},

date = {2021-03-00},

urldate = {2021-03-00},

journal = {Microbial Pathogenesis},

volume = {152},

publisher = {Elsevier BV},

abstract = {The lectins are carbohydrate-binding proteins that are highly specific to sugar groups associated to other molecules. In addition to interacting with carbohydrates, a number of studies have reported the ability of these proteins to modulate the activity of several antibiotics against multidrug-resistant (MDR) strains. In this study, we report the enhanced antibacterial activity of the gentamicin against MDR strains when complexed with a lectin from Canavalia ensiformis seeds (ConA). Hemagglutination activity test and intrinsic fluorescence spectroscopy revealed that the gentamicin can interact with ConA most likely via the carbohydrate recognition domain (CRD) with binding constant (Kb) value estimated of (0.44 ± 0.04) x 104 M−1. Furthermore, the minimum inhibitory concentrations (MIC) obtained for ConA against all strains studied were not clinically relevant (MIC ≥ 1024 μg/mL). However, when ConA was combined with gentamicin, a significant increase in antibiotic activity was observed against Staphylococcus aureus and Escherichia coli. The present study showed that ConA has an affinity for gentamicin and modulates its activity against MDR strains. These results indicate that ConA improves gentamicin performance and is a promising candidate for structure/function analyses.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{FrancoCairo2021,

title = {On the roles of AA15 lytic polysaccharide monooxygenases derived from the termite Coptotermes gestroi},

author = {João Paulo L. Franco Cairo and David Cannella and Leandro C. Oliveira and Thiago A. Gonçalves and Marcelo V. Rubio and Cesar R.F. Terrasan and Robson Tramontina and Luciana S. Mofatto and Marcelo F. Carazzolle and Wanius Garcia and Claus Felby and André Damasio and Paul H. Walton and Fabio Squina},

url = {https://www.sciencedirect.com/science/article/pii/S0162013420303445},

doi = {10.1016/j.jinorgbio.2020.111316},

issn = {0162-0134},

year  = {2021},

date = {2021-03-00},

urldate = {2021-03-00},

journal = {Journal of Inorganic Biochemistry},

volume = {216},

publisher = {Elsevier BV},

abstract = {Lytic polysaccharide monooxygenases (LPMOs) are copper-dependent enzymes which catalyze the oxidative cleavage of polysaccharides. LPMOs belonging to family 15 in the Auxiliary Activity (AA) class from the Carbohydrate-Active Enzyme database are found widespread across the Tree of Life, including viruses, algae, oomycetes and animals. Recently, two AA15s from the firebrat Thermobia domestica were reported to have oxidative activity, one towards cellulose or chitin and the other towards chitin, signalling that AA15 LPMOs from insects potentially have different biochemical functions. Herein, we report the identification and characterization of two family AA15 members from the lower termite Coptotermes gestroi. Addition of Cu(II) to CgAA15a or CgAA15b had a thermostabilizing effect on both. Using ascorbate and O2 as co-substrates, CgAA15a and CgAA15b were able to oxidize chitin, but showed no activity on celluloses, xylan, xyloglucan and starch. Structural models indicate that the LPMOs from C. gestroi (CgAA15a/CgAA15b) have a similar fold but exhibit key differences in the catalytic site residues when compared to the cellulose/chitin-active LPMO from T. domestica (TdAA15a), especially the presence of a non-coordinating phenylalanine nearby the Cu ion in CgAA15a/b, which appears as a tyrosine in the active site of TdAA15a. Despite the overall similarity in protein folds, however, mutation of the active site phenylalanine in CgAA15a to a tyrosine did not expanded the enzymatic specificity from chitin to cellulose. Our data show that CgAA15a/b enzymes are likely not involved in lignocellulose digestion but might play a role in termite developmental processes as well as on chitin and nitrogen metabolisms.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{BomedianoCamillo2021,

title = {Structural modelling and thermostability of a serine protease inhibitor belonging to the Kunitz-BPTI family from the Rhipicephalus microplus tick},

author = {Lívia de Moraes Bomediano Camillo and Graziele Cristina Ferreira and Adriana Feliciano Alves Duran and Flavia Ribeiro Santos da Silva and Wanius Garcia and Ana Lígia Scott and Sergio Daishi Sasaki},

url = {https://www.sciencedirect.com/science/article/pii/S0300908420303369},

doi = {10.1016/j.biochi.2020.12.014},

issn = {0300-9084},

year  = {2021},

date = {2021-02-00},

urldate = {2021-02-00},

journal = {Biochimie},

volume = {181},

pages = {226--233},

publisher = {Elsevier BV},

abstract = {rBmTI-A is a recombinant serine protease inhibitor that belongs to the Kunitz-BPTI family and that was cloned from Rhipicephalus microplus tick. rBmTI-A has inhibitory activities on bovine trypsin, human plasma kallikrein, human neutrophil elastase and plasmin with dissociation constants in nM range. It is characterized by two inhibitory domains and each domain presents six cysteines that form three disulfide bonds, which contribute to the high stability of its structure. Previous studies suggest that serine protease inhibitor rBmTI-A has a protective potential against pulmonary emphysema in mice and anti-inflammatory potential. Besides that, rBmTI-A presented a potent inhibitory activity against in vitro vessel formation. In this study, the tertiary structure of rBmTI-A was modeled. The structure stabilization was evaluated by molecular dynamics analysis. Circular dichroism spectroscopy data corroborated the secondary structure found by the homology modelling. Also, in circular dichroism data it was shown a thermostability of rBmTI-A until approximately 70 °C, corroborated by inhibitory assays toward trypsin.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}