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1.
Ecotoxicol Environ Saf ; 274: 116201, 2024 Apr 01.
Artigo em Inglês | MEDLINE | ID: mdl-38489901

RESUMO

Seafood products are globally consumed, and there is an increasing demand for the quality and safety of these products among consumers. Some seafoods are easily contaminated by marine biotoxins in natural environments or cultured farming processes. When humans ingest different toxins accumulated in seafood, they may exhibit different poisoning symptoms. According to the investigations, marine toxins produced by harmful algal blooms and various other marine organisms mainly accumulate in the body organs such as liver and digestive tract of seafood animals. Several regions around the world have reported incidents of seafood poisoning by biotoxins, posing a threat to human health. Thus, most countries have legislated to specify the permissible levels of these biotoxins in seafood. Therefore, it is necessary for seafood producers and suppliers to conduct necessary testing of toxins in seafood before and after harvesting to prohibit excessive toxins containing seafood from entering the market, which therefore can reduce the occurrence of seafood poisoning incidents. In recent years, some technologies which can quickly, conveniently, and sensitively detect biological toxins in seafood, have been developed and validated, these technologies have the potential to help seafood producers, suppliers and regulatory authorities. This article reviews the seafood toxins sources and types, mechanism of action and bioaccumulation of marine toxins, as well as legislation and rapid detection technologies for biotoxins in seafood for official and fishermen supervision.


Assuntos
Doenças Transmitidas por Alimentos , Toxinas Marinhas , Animais , Humanos , Toxinas Marinhas/toxicidade , Alimentos Marinhos/análise , Bioacumulação , Doenças Transmitidas por Alimentos/epidemiologia , Proliferação Nociva de Algas
2.
Appl Environ Microbiol ; 89(3): e0218622, 2023 03 29.
Artigo em Inglês | MEDLINE | ID: mdl-36847541

RESUMO

Cytochrome P450 enzymes are promising biocatalysts for industrial use because they catalyze site-selective C-H oxidation and have diverse catalytic reactions and a broad substrate range. In this study, the 2α-hydroxylation activity of CYP154C2 from Streptomyces avermitilis MA-4680T toward androstenedione (ASD) was identified by an in vitro conversion assay. The testosterone (TES)-bound structure of CYP154C2 was solved at 1.42 Å, and this structure was used to design eight mutants, including single, double, and triple mutants, to improve the conversion efficiency. Mutants L88F/M191F and M191F/V285L were found to enhance the conversion rates significantly (i.e., 8.9-fold and 7.4-fold for TES, 46.5-fold and 19.5-fold for ASD, respectively) compared with the wild-type (WT) enzyme while retaining high 2α-position selectivity. The substrate binding affinity of the L88F/M191F mutant toward TES and ASD was enhanced compared with that of WT CYP154C2, supporting the measured increase in the conversion efficiencies. Moreover, the total turnover number and kcat/Km of the L88F/M191F and M191F/V285L mutants increased significantly. Interestingly, all mutants containing L88F generated 16α-hydroxylation products, suggesting that L88 in CYP154C2 plays a vital role in substrate selectivity and that the amino acid corresponding to L88 in the 154C subfamily affects the orientation of steroid binding and substrate selectivity. IMPORTANCE Hydroxylated derivatives of steroids play essential roles in medicine. Cytochrome P450 enzymes selectively hydroxylate methyne groups on steroids, which can dramatically change their polarity, biological activity and toxicity. There is a paucity of reports on the 2α-hydroxylation of steroids, and documented 2α-hydroxylate P450s show extremely low conversion efficiency and/or low regio- and stereoselectivity. This study conducted crystal structure analysis and structure-guided rational engineering of CYP154C2 and efficiently enhanced the conversion efficiency of TES and ASD with high regio- and stereoselectivity. Our results provide an effective strategy and theoretical basis for the 2α-hydroxylation of steroids, and the structure-guided rational design of P450s should facilitate P450 applications in the biosynthesis of steroid drugs.


Assuntos
Sistema Enzimático do Citocromo P-450 , Esteroides , Hidroxilação , Esteroides/metabolismo , Sistema Enzimático do Citocromo P-450/metabolismo , Oxirredução , Testosterona/metabolismo , Especificidade por Substrato
3.
Mar Drugs ; 21(3)2023 Feb 25.
Artigo em Inglês | MEDLINE | ID: mdl-36976202

RESUMO

In recent years, allergic diseases have occurred frequently, affecting more than 20% of the global population. The current first-line treatment of anti-allergic drugs mainly includes topical corticosteroids, as well as adjuvant treatment of antihistamine drugs, which have adverse side effects and drug resistance after long-term use. Therefore, it is essential to find alternative anti-allergic agents from natural products. High pressure, low temperature, and low/lack of light lead to highly functionalized and diverse functional natural products in the marine environment. This review summarizes the information on anti-allergic secondary metabolites with a variety of chemical structures such as polyphenols, alkaloids, terpenoids, steroids, and peptides, obtained mainly from fungi, bacteria, macroalgae, sponges, mollusks, and fish. Molecular docking simulation is applied by MOE to further reveal the potential mechanism for some representative marine anti-allergic natural products to target the H1 receptor. This review may not only provide insight into information about the structures and anti-allergic activities of natural products from marine organisms but also provides a valuable reference for marine natural products with immunomodulatory activities.


Assuntos
Antialérgicos , Produtos Biológicos , Animais , Organismos Aquáticos/química , Antialérgicos/farmacologia , Produtos Biológicos/química , Simulação de Acoplamento Molecular , Fungos/química
4.
Chem Biodivers ; 20(8): e202300831, 2023 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-37349894

RESUMO

Bacterial infected environments and resulting bacterial infections have been threatening the human health globally. Due to increased bacterial resistance caused by improper and excessive use of antibiotics, antibacterial biomaterials are being developed as alternatives to antibiotics in some cases. Herein, an advanced multifunctional hydrogel with excellent antibacterial properties, enhanced mechanical properties, biocompatibility and self-healing performance, was designed through freezing-thawing method. This hydrogel network is composed of polyvinyl alcohol (PVA), carboxymethyl chitosan (CMCS), protocatechualdehyde (PA), ferric iron (Fe) and an antimicrobial cyclic peptide actinomycin X2 (Ac.X2). The double dynamic bonds among protocatechualdehyde (PA), ferric iron (Fe) and carboxymethyl chitosan containing coordinate bond (catechol-Fe) as well as dynamic Schiff base bonds and hydrogen bonds endowed the hydrogel with enhanced mechanical properties. Successful formation of hydrogel was confirmed through ATR-IR and XRD, and structural evaluation through SEM analysis, whereas mechanical properties were tested with electromechanical universal testing machine. The resulting PVA/CMCS/Ac.X2/PA@Fe (PCXPA) hydrogel has favorable biocompatibility and excellent broad-spectrum antimicrobial activity against both S. aureus (95.3 %) and E. coli (90.2 %) compared with free-soluble Ac.X2, which exhibited subpar performance against E. coli reported in our previous studies. This work provides a new insight on preparing multifunctional hydrogels containing antimicrobial peptides as antibacterial material.


Assuntos
Anti-Infecciosos , Quitosana , Humanos , Quitosana/química , Peptídeos Cíclicos , Álcool de Polivinil/química , Escherichia coli , Staphylococcus aureus , Antibacterianos/farmacologia , Antibacterianos/química , Hidrogéis/farmacologia , Hidrogéis/química , Catecóis , Ferro
5.
Chem Biodivers ; 20(6): e202300445, 2023 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-37148532

RESUMO

Development of suitable antimicrobial biomaterials for hygienic wound dressing and healing is an important requirement for medical application. Durable mechanical properties increase the application range of biomaterial in different environmental and biological conditions. Due to the inherent brittleness of silk fibroin (SF), polyurethane fiber (PUF) was used to modify SF containing actinomycin X2 (Ac.X2) to prepare silk fibroin@actinomycin X2 /polyurethane fiber (ASF/PUF) blend membranes. The ASF/PUF blend membrane was developed by solution casting method. Incorporation of PUF improved the flexibility of material and introduction of Ac.X2 has increased antibacterial activity of materials. Excellent mechanical properties (tensile strength up to 25.7 MPa and elongation at break up to 946.5 %) of 50 % SF+50 % PUF blend membrane were proved by tensile testing machine. FT-IR spectra, TGA, contact angle and DMA were tested to prove the blend membrane's physico-chemical characteristics. ASF/PUF blend membrane displayed satisfactory antibacterial activity against S. aureus, and the cytotoxicity tests showed that the blend membrane has better biosafety compared to directly applied Ac.X2 in soluble form. These results suggest that the modification of SF through PUF for development of flexible antibacterial membranes has great potential application value in the field of silk-like material fabrication.


Assuntos
Fibroínas , Fibroínas/farmacologia , Fibroínas/química , Poliuretanos/farmacologia , Poliuretanos/química , Espectroscopia de Infravermelho com Transformada de Fourier , Staphylococcus aureus , Seda/química , Materiais Biocompatíveis/química , Antibacterianos/farmacologia
6.
Int J Mol Sci ; 24(7)2023 Mar 27.
Artigo em Inglês | MEDLINE | ID: mdl-37047243

RESUMO

Actinomycin is a family of chromogenic lactone peptides that differ in their peptide portions of the molecule. An antimicrobial peptide, actinomycin X2 (Ac.X2), was produced through the fermentation of a Streptomyces cyaneofuscatus strain. Immobilization of Ac.X2 onto a prepared silk fibroin (SF) film was done through a carbodiimide reaction. The physical properties of immobilized Ac.X2 (antimicrobial films, AMFs) were analyzed by ATR-FTIR, SEM, AFM, and WCA. The findings from an in vitro study showed that AMFs had a more broad-spectrum antibacterial activity against both S. aureus and E. coli compared with free Ac.X2, which showed no apparent strong effect against E. coli. These AMFs showed a suitable degradation rate, good hemocompatibility, and reduced cytotoxicity in the biocompatibility assay. The results of in vivo bacterially infected wound healing experiments indicated that wound inflammation was prevented by AMFs, which promoted wound repair and improved the wound microenvironment. This study revealed that Ac.X2 transformation is a potential candidate for skin wound healing.


Assuntos
Peptídeos Antimicrobianos , Dactinomicina , Fibroínas , Proteínas Imobilizadas , Cicatrização , Dactinomicina/química , Dactinomicina/farmacologia , Fibroínas/química , Fibroínas/farmacologia , Proteínas Imobilizadas/química , Proteínas Imobilizadas/farmacologia , Cicatrização/efeitos dos fármacos , Streptomyces/metabolismo , Escherichia coli/efeitos dos fármacos , Staphylococcus aureus/efeitos dos fármacos , Espectroscopia de Infravermelho com Transformada de Fourier , Microscopia de Força Atômica , Fermentação , Teste de Materiais , Materiais Biocompatíveis/química , Materiais Biocompatíveis/farmacologia , Animais , Ratos , Masculino , Ratos Sprague-Dawley
7.
Molecules ; 28(6)2023 Mar 20.
Artigo em Inglês | MEDLINE | ID: mdl-36985758

RESUMO

Two aplysiatoxin derivatives, neo-debromoaplysiatoxin I (1) and neo-debromoaplysiatoxin J (2), were isolated from marine cyanobacterium Lyngbya sp. collected from the South China Sea. Their structures including absolute configurations were assigned by spectroscopic analysis, in combination with GIAO NMR shift calculation and DP4+ analysis. Structures of neo-debromoaplysiatoxin I and neo-debromoaplysiatoxin J contained a decahydro-5H-pyrano [2,3,4-de] chromen-5-one 6/6/6 ring skeleton and an intriguing peroxide bridge group, respectively, which are unprecedented structure scaffold and motif in aplysiatoxins. Two compounds displayed comparable inhibitory activities against Kv1.5 K+ channel with IC50 values of 2.59 ± 0.37 µM (1) and 1.64 ± 0.15 µM (2); however, they presented differential cytotoxic effects. It is worth noting that neo-debromoaplysiatoxin J, containing a peroxide bridge, showed remarkable cytotoxicity against four cancer cell lines including SW480, SGC7901, LoVo and PC-9 compared to the human normal cell line.


Assuntos
Cianobactérias , Lyngbya , Humanos , Canal de Potássio Kv1.5 , Cianobactérias/química , Toxinas de Lyngbya/química , Linhagem Celular , Estrutura Molecular
8.
Int J Syst Evol Microbiol ; 72(12)2022 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-36748603

RESUMO

A Gram-stain-negative, aerobic, chemoheterotrophic and rod-shaped strain, designated as C5T, was isolated from intertidal surface seawater in Taizhou, Zhejiang Province, PR China and characterized using a polyphasic taxonomic approach. Strain C5T could produce carotenoids and bacteriochlorophyll a. Growth was observed at 20-45 °C, at pH 6.0-9.0 and with 0-8.0 % (w/v) NaCl. The 16S rRNA gene sequence identity analysis revealed that strain C5T was the most closely related to Qipengyuania nanhaisediminis CGMCC 1.7715T (98.8%) and Erythrobacter litoralis DSM 8509T (98.7%). The phylogenetic reconstruction based on core genes demonstrated that strain C5T was clustered into the members of the genus Erythrobacter. The average nucleotide identity and digital DNA-DNA hybridization values between strain C5T and Erythrobacter type strains were lower than 76 and 25 %, respectively. The predominant and minor respiratory quinones were identified as ubiquinone-10 and ubiquinone-9. The major fatty acids were summed feature 8 (C18 : 1 ω7c and/or C18 : 1 ω6c) and iso-C18 : 0. Polar lipids included phosphatidylethanolamine, phosphatidylglycerol, a glycosphingolipid and an unidentified aminolipid. Based on the genetic, chemotaxonomic and phenotypic data, strain C5T is concluded to represent a novel species in the genus Erythrobacter, for which the name Erythrobacter aurantius sp. nov. is proposed. The type strain is C5T (=MCCC 1K05108T=KCTC 92307T).


Assuntos
Ácidos Graxos , Sphingomonadaceae , Ácidos Graxos/química , Fosfolipídeos/química , Filogenia , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Composição de Bases , DNA Bacteriano/genética , Técnicas de Tipagem Bacteriana , Água do Mar
9.
Chem Biodivers ; 19(5): e202200177, 2022 May.
Artigo em Inglês | MEDLINE | ID: mdl-35426465

RESUMO

Cytochrome P450 enzymes (CYPs or P450s) are ubiquitous heme-dependent enzymes that catalyze the monooxygenation of non-activated C-H bonds to modify the structure of the substrate. In this study, we heterologously expressed CYP107X1 from Streptomyces avermitilis and conducted in vitro substrate screening using the alternative redox partners putidaredoxin and putidaredoxin reductase. CYP107X1 catalyzed the 16α-hydroxylation of progesterone with regio- and stereoselectivity. The spectroscopic analyses showed that CYP107X1 bound progesterone with a relatively high Kd value of 65.3±38.9 µM. The Km and kcat values for progesterone were estimated to be 47.7±12.0 µM and 0.30 min-1 , respectively. Furthermore, a crystal structure was obtained of CYP107X1 bound with glycerol from the buffer solution. Interestingly, a conserved threonine was replaced with asparagine in CYP107X1, indicating that it may adopt an unnatural proton transfer process and play a crucial role in its catalytic activity.


Assuntos
Progesterona , Streptomyces , Sistema Enzimático do Citocromo P-450/metabolismo , Hidroxilação , Progesterona/metabolismo , Streptomyces/metabolismo
10.
Arch Microbiol ; 204(1): 86, 2021 Dec 27.
Artigo em Inglês | MEDLINE | ID: mdl-34961889

RESUMO

Three yellow-pigmented, Gram-stain-negative, aerobic, motile by flagella and rod-shaped strains, designated as MCT, PC and RC, were isolated from stems of Populus euphratica. Growth of those three strains occurs at 4-40 °C, pH 6.0-10.0 and with 0.5-18.0% (w/v) NaCl. Respiratory quinones contained ubiquinone-9 and ubiquione-8 as major and minor components, respectively. Major fatty acids (> 10%) were summed feature 8 (C18:1ω6c and/or C18:1ω7c), summed feature 3 (C16:1ω6c and/or C16:1ω7c) and C16:0. Polar lipids included diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, two unidentified phospholipids, one unidentified aminolipid, one unidentified glycolipid and four unidentified lipids. Strains MCT, PC and RC shared pairwise 16S rRNA gene sequence similarities of 99.9-100.0%, and showed higher similarities of 98.4-98.5% with Halomonas songnenensis NEAU-ST10-39T and 98.3-98.4% with Halomonas nanhaiensis YIM M 13059T than to other Halomonas type strains. Genomic comparisons revealed that those three strains had the pan-genome consisting of 4446 orthologous clusters, among which 676 orthologous clusters were absent in other Halomonas type strains. Phylogenomic tree indicated that strains MCT, PC and RC formed an independently stable clade with Halomonas nanhaiensis YIM M 13059T and Halomonas songnenensis NEAU-ST10-39T. The average nucleotide identity and digital DNA-DNA hybridization values between those three strains and other Halomonas type strains were < 89.9% and < 39.3%, respectively. Based upon phenotypic, chemotaxonomic, phylogenetic and genomic results, strains MCT, PC and RC represent a novel species in the genus Halomonas, for which the name Halomonas populi sp. nov. is proposed. The type strain is MCT (= JCM 33545T = MCCC 1K03942T).


Assuntos
Halomonas , Populus , Técnicas de Tipagem Bacteriana , Composição de Bases , DNA Bacteriano/genética , Ácidos Graxos/análise , Halomonas/genética , Fosfolipídeos/análise , Filogenia , RNA Ribossômico 16S/genética , Análise de Sequência de DNA
11.
Arch Microbiol ; 203(8): 5133-5139, 2021 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-34319420

RESUMO

A Marinomonas-like, Gram-stain-negative, strictly aerobic and rod to ovoid-shaped bacterium, designated as strain A79T, was isolated from the seawater mixtures of oyster shells and brown algae in a coastal intertidal zone of Zhoushan, China. The strain was positive for oxidase and catalase. Colonies grown on marine agar for 48 h were round, milky white, smooth and moist with the diameter of 2-3 mm. Growth was observed at 15-30 °C (optimum, 25℃), pH 5.5-9.5 (optimum, pH 8.5) and with 0.5-8% (w/v) NaCl (optimum, 2-2.5%). The G + C content based on the genome sequence was 46.0%. The only respiratory quinone was Q-8. The main polar lipids contained phosphatidylglycerol, phosphatidylethanolamine, unidentified glycolipids, unidentified phospholipid and three unidentified lipids. The major fatty acids (> 10%) were C16:0, Summed feature 3 (comprising C16:1 ω6c and/or C16:1 ω7c) and summed feature 8 (comprising C18:1 ω6c and/or C18:1 ω7c). The 16S rRNA gene sequence similarity between strain A79T and Marinomonas pollencensis IVIA-Po-185T was 97.4%, the similarities with other type strains of the genus Marinomonas were 93.8-96.7%. Based on the results, Marinomonas vulgaris sp. nov. was proposed as a novel species. The type strain is A79T (= MCCC 1K05799T = KCTC 82519T = JCM 34473T).


Assuntos
Marinomonas , Técnicas de Tipagem Bacteriana , Composição de Bases , DNA Bacteriano/genética , Ácidos Graxos , Marinomonas/genética , Hibridização de Ácido Nucleico , Fosfolipídeos , Filogenia , RNA Ribossômico 16S/genética , Água do Mar , Análise de Sequência de DNA
12.
Mar Drugs ; 19(11)2021 Nov 10.
Artigo em Inglês | MEDLINE | ID: mdl-34822500

RESUMO

The actinomycetes have proven to be a rich source of bioactive secondary metabolites and play a critical role in the development of pharmaceutical researches. With interactions of host organisms and having special ecological status, the actinomycetes associated with marine animals, marine plants, macroalgae, cyanobacteria, and lichens have more potential to produce active metabolites acting as chemical defenses to protect the host from predators as well as microbial infection. This review focuses on 536 secondary metabolites (SMs) from actinomycetes associated with these marine organisms covering the literature to mid-2021, which will highlight the taxonomic diversity of actinomycetes and the structural classes, biological activities of SMs. Among all the actinomycetes listed, members of Streptomyces (68%), Micromonospora (6%), and Nocardiopsis (3%) are dominant producers of secondary metabolites. Additionally, alkaloids (37%), polyketides (33%), and peptides (15%) comprise the largest proportion of natural products with mostly antimicrobial activity and cytotoxicity. Furthermore, the data analysis and clinical information of SMs have been summarized in this article, suggesting that some of these actinomycetes with multiple host organisms deserve more attention to their special ecological status and genetic factors.


Assuntos
Actinobacteria , Antibacterianos/farmacologia , Animais , Antibacterianos/química , Organismos Aquáticos , Produtos Biológicos , Humanos
13.
Mar Drugs ; 19(11)2021 Nov 11.
Artigo em Inglês | MEDLINE | ID: mdl-34822501

RESUMO

Potassium channel Kv1.5 has been considered a key target for new treatments of atrial tachyarrhythmias, with few side effects. Four new debromoaplysiatoxin analogues with a 6/6/12 fused ring system were isolated from marine cyanobacterium Lyngbya sp. Their planar structures were elucidated by HRESIMS, 1D and 2D NMR. The absolute configuration of oscillatoxin J (1) was determined by single-crystal X-ray diffraction, and the absolute configurations of oscillatoxin K (2), oscillatoxin L (3) and oscillatoxin M (4) were confirmed on the basis of GIAO NMR shift calculation followed by DP4 analysis. The current study confirmed the absolute configuration of the pivotal chiral positions (7S, 9S, 10S, 11R, 12S, 15S, 29R and 30R) at traditional ATXs with 6/12/6 tricyclic ring system. Compound 1, 2 and 4 exhibited blocking activities against Kv1.5 with IC50 values of 2.61 ± 0.91 µM, 3.86 ± 1.03 µM and 3.79 ± 1.01 µM, respectively. However, compound 3 exhibited a minimum effect on Kv1.5 at 10 µM. Furthermore, all of these new debromoaplysiatoxin analogs displayed no apparent activity in a brine shrimp toxicity assay.


Assuntos
Canal de Potássio Kv1.5/efeitos dos fármacos , Toxinas de Lyngbya/farmacologia , Lyngbya , Animais , Organismos Aquáticos , Artemia , Humanos , Concentração Inibidora 50 , Canal de Potássio Kv1.5/antagonistas & inibidores , Toxinas de Lyngbya/química , Camundongos , Relação Estrutura-Atividade
14.
Mar Drugs ; 20(1)2021 Dec 23.
Artigo em Inglês | MEDLINE | ID: mdl-35049871

RESUMO

Actinomycins as clinical medicine have been extensively studied, while few investigations were conducted to discover the feasibility of actinomycins as antimicrobial natural dye contributing to the medical value of the functional fabrics. This study was focused on the application of actinomycin X2 (Ac.X2), a peptide pigment cultured from marine-derived Streptomyces cyaneofuscatus, in the dyeing and finishing of silk fabric. The dyeing potential of Ac.X2 with silk vs. cotton fabrics was assessed. As a result, the silk fabric exhibited greater uptake and color fastness with Ac.X2. Through Fourier transform infrared spectroscopy (FT-IR), scanning electron microscopy (SEM), and X-ray diffraction (XRD) analyses, some changes of chemical property for the dyed fabric and Ac.X2 were studied. The silk fabric dyed with Ac.X2 exhibited good UV protection ability. The antibacterial properties of dyed and finished silk were also evaluated, which exhibited over 90% antibacterial activity even after 20 washing cycles. In addition, the brine shrimp assay was conducted to evaluate the general toxicity of the tested fabric, and the results indicated that the dyed silk fabrics had a good biological safety property.


Assuntos
Antibacterianos/farmacologia , Corantes/farmacologia , Dactinomicina/análogos & derivados , Streptomyces , Animais , Antibacterianos/química , Organismos Aquáticos , Artemia/efeitos dos fármacos , Corantes/química , Dactinomicina/química , Dactinomicina/farmacologia , Testes de Sensibilidade Microbiana , Microscopia Eletrônica de Varredura , Seda , Espectroscopia de Infravermelho com Transformada de Fourier , Staphylococcus aureus/efeitos dos fármacos
15.
Chem Biodivers ; 18(4): e2000910, 2021 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-33656282

RESUMO

Cytochrome P450 enzymes (P450s) are versatile biocatalysts, which insert a molecular oxygen into inactivated C-H bonds under mild conditions. CYP105D7 from Streptomyces avermitilis has been reported as a bacterial substrate-promiscuous P450 which catalyzes the hydroxylation of 1-deoxypentalenic acid, diclofenac, naringenin, compactin and steroids. In this study, CYP105D7 catalyzes hydroxylation, epoxidation and dehydrogenation of capsaicin, a pharmaceutical agent, revealing its functional diversity. The kinetic parameters of the CYP105D7 oxidation of capsaicin were determined as Km =311.60±87.30 µM and kcat =2.01±0.33 min-1 . In addition, we conducted molecular docking, mutagenesis and substrate binding analysis, indicating that Arg81 plays crucial role in the capsaicin binding and catalysis. To our best knowledge, this study presents the first report to illustrate that capsaicin can be catalyzed by prokaryotic P450s.


Assuntos
Capsaicina/metabolismo , Sistema Enzimático do Citocromo P-450/metabolismo , Streptomyces/enzimologia , Biocatálise , Capsaicina/química , Hidrogenação , Hidroxilação , Simulação de Acoplamento Molecular , Estrutura Molecular
16.
J Nat Prod ; 83(9): 2647-2654, 2020 09 25.
Artigo em Inglês | MEDLINE | ID: mdl-32816473

RESUMO

Eight new dioxopiperazine alkaloids, penispirozines A-H (1-8), were discovered from the mangrove-derived fungus Penicillium janthinellum HDN13-309. Their structures were elucidated by spectroscopic analysis, TDDFT-ECD calculations, and X-ray diffraction. Compound 1 had an unusual pyrazino[1,2]oxazadecaline coupled with a thiophane ring system, and compound 2 possessed a 6/5/6/5/6 pentacyclic ring system with two rare spirocyclic centers. Interestingly, compounds 3-8 were distinguished by not only the existence of a spiro-thiophane or spiro-furan ring system but also the chirality of the pentacyclic moiety. Compounds 3 and 4 increased the expression of the two relevant phase II detoxifying enzymes SOD2 and HO-1 at 10 µM.


Assuntos
Alcaloides/química , Avicennia/microbiologia , Penicillium/química , Cristalografia por Raios X , Indução Enzimática/efeitos dos fármacos , Fermentação , Heme Oxigenase-1/biossíntese , Espectroscopia de Ressonância Magnética , Estrutura Molecular , Ressonância Magnética Nuclear Biomolecular , Superóxido Dismutase/biossíntese , Difração de Raios X
17.
Fish Shellfish Immunol ; 84: 704-710, 2019 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-30359751

RESUMO

Japanese eel (Anguilla japonica) has become a commercially important fish species all over the world. High-density aquaculture has led to congestion and contributed to bacterial infection outbreaks that have caused high mortality. Therefore a 56-days feeding trial was conducted to determine the effects of dietary Bacillus amyloliquefaciens (GB-9) and Yarrowia lipolytica lipase2 (YLL2) on growth performance, digestive enzymes activity, innate immunity and resistance to pathogens of A. japonica. Fish growth performance was significantly affected by dietary YLL2 supplementation but not by GB-9. Fish fed diets with YLL2 at 2.0 g/kg diet in combination of high and low levels of GB-9 (5.0 g/kg and 2.0 g/kg) produced the highest growth. For digestive enzyme, lipase and trypsin activities was promoted by dietary containing YLL2, while amylase activities was increased by dietary containing YLL2, GB-9 single or combination. For innate immunity, the mucus lysozyme activity, leukocytes phagocytosis activity and reactive oxygen species level of skin, peroxidase and lysozyme activity of serum were enhanced in fish fed with GB-9 compared to those in control group (p < 0.05). The highest resistance to Vibrio anguillarum and Aeromonas hydrophila was determined in fish fed with 5.0 g kg-1 GB-9 + 2.0 g/kg YLL2. This study demonstrated that GB-9 and YLL2 enhanced non-specific immune defense system of A. japonica, providing them with higher resistance to pathogens. The present results suggested that the combination of these supplements could be considered as potential biological additives for aquaculture farmed fish.


Assuntos
Anguilla/imunologia , Bacillus amyloliquefaciens/química , Hidrolases de Éster Carboxílico/administração & dosagem , Doenças dos Peixes/imunologia , Proteínas Fúngicas/administração & dosagem , Imunidade Inata/efeitos dos fármacos , Imunidade nas Mucosas/efeitos dos fármacos , Probióticos/farmacologia , Aeromonas hydrophila/fisiologia , Anguilla/crescimento & desenvolvimento , Anguilla/metabolismo , Animais , Dieta/veterinária , Suplementos Nutricionais/análise , Relação Dose-Resposta a Droga , Trato Gastrointestinal/enzimologia , Infecções por Bactérias Gram-Negativas/imunologia , Infecções por Bactérias Gram-Negativas/veterinária , Distribuição Aleatória , Vibrio/fisiologia , Vibrioses/imunologia , Vibrioses/veterinária
18.
Mar Drugs ; 17(12)2019 Nov 21.
Artigo em Inglês | MEDLINE | ID: mdl-31766406

RESUMO

A pair of stereoisomers possessing novel structures with 6/6/5 fused-ring systems, neo-debromoaplysiatoxin E (1) and neo-debromoaplysiatoxin F (2), were isolated from the marine cyanobacterium Lyngbya sp. Their structures were elucidated using various spectroscopic techniques including high resolution electrospray ionization mass spectroscopy (HRESIMS) and nuclear magnetic resonance (NMR). The absolute stereochemistry was determined by calculated electronic circular dichroism (ECD) and gauge-independent atomic orbital (GIAO) NMR shift calculation followed by DP4+ analysis. Significantly, this is the first report on aplysiatoxin derivatives with different absolute configurations at C9-C12 (1: 9S, 10R, 11S, 12S; 2: 9R, 10S, 11R, 12R). Compounds 1 and 2 exhibited potent blocking activities against Kv1.5 with IC50 values of 1.22 ± 0.22 µM and 2.85 ± 0.29 µM, respectively.


Assuntos
Organismos Aquáticos/química , Cianobactérias/química , Canal de Potássio Kv1.5/antagonistas & inibidores , Toxinas de Lyngbya/farmacologia , Animais , Células CHO , Dicroísmo Circular , Cricetulus , Canal de Potássio Kv1.5/metabolismo , Toxinas de Lyngbya/química , Toxinas de Lyngbya/isolamento & purificação , Espectroscopia de Ressonância Magnética , Estrutura Molecular , Estereoisomerismo
19.
Mar Drugs ; 17(9)2019 Aug 26.
Artigo em Inglês | MEDLINE | ID: mdl-31454987

RESUMO

Bacterial secondary metabolites have huge application potential in multiple industries. Biosynthesis of bacterial secondary metabolites are commonly encoded in a set of genes that are organized in the secondary metabolism biosynthetic gene clusters (SMBGCs). The development of genome sequencing technology facilitates mining bacterial SMBGCs. Marine Streptomyces is a valuable resource of bacterial secondary metabolites. In this study, 87 marine Streptomyces genomes were obtained and carried out into comparative genomic analysis, which revealed their high genetic diversity due to pan-genomes owning 123,302 orthologous clusters. Phylogenomic analysis indicated that the majority of Marine Streptomyces were classified into three clades named Clade I, II, and III, containing 23, 38, and 22 strains, respectively. Genomic annotations revealed that SMBGCs in the genomes of marine Streptomyces ranged from 16 to 84. Statistical analysis pointed out that phylotypes and ecotypes were both associated with SMBGCs distribution patterns. The Clade I and marine sediment-derived Streptomyces harbored more specific SMBGCs, which consisted of several common ones; whereas the Clade II and marine invertebrate-derived Streptomyces have more SMBGCs, acting as more plentiful resources for mining secondary metabolites. This study is beneficial for broadening our knowledge about SMBGC distribution patterns in marine Streptomyces and developing their secondary metabolites in the future.


Assuntos
Organismos Aquáticos/genética , Genes Bacterianos , Família Multigênica , Metabolismo Secundário/genética , Streptomyces/genética , Organismos Aquáticos/metabolismo , Produtos Biológicos/metabolismo , Vias Biossintéticas/genética , Genômica , Filogenia , Streptomyces/metabolismo
20.
Fish Shellfish Immunol ; 82: 250-257, 2018 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-30125703

RESUMO

A 12-weeks feeding trial was performed to investigate the possible effects of supplementation of Hybrid sturgeon diet with Bacillus amyloliquefaciens (GB-9) and Yarrowia lipolytica lipase2 (YLL2) single or combined on immune response and growth performance of Hybrid sturgeon (Acipenser schrenkii ♂and Acipenser baeri ♀). For this aim, Hybrid sturgeons were fed with four experimental diets namely: Diet 1 (0-control), Diet 2 (5.0 g/kg GB-9), Diet 3 (4.0 g/kg YLL2), and Diet 4 (5.0 g/kg GB-9 + 4.0 g/kg YLL2), respectively. After fed with varied diets, growth performance, mucosal immune response, leukocytes immune response and serum immunological response were measured. The results indicated that supplementations of GB-9 + YLL2 resulted in a significant increase in final weight, Docosahexaenoic acid (DHA) and Eicosapentenoic acid (EPA) concentration, compared with that of control (p < 0.05). For innate immunity, the results showed that skin mucus lysozyme activity, leukocytes phagocytosis activity and reactive oxygen species level, and serum alternative complement pathway activity, peroxidase and lysozyme activity were significantly higher in supplemented groups compared to the control (p < 0.05). The highest values were recorded in fish fed both YLL2 and GB-9 with respect to the individual application. The present results suggested that the combination of these supplementation could be considered as potential feed-additives for aquaculture farmed fish.


Assuntos
Bacillus amyloliquefaciens/química , Hidrolases de Éster Carboxílico/administração & dosagem , Peixes/crescimento & desenvolvimento , Peixes/imunologia , Proteínas Fúngicas/administração & dosagem , Probióticos/farmacologia , Ração Animal/análise , Animais , Cruzamento , Dieta/veterinária , Suplementos Nutricionais/análise , Imunidade Inata/efeitos dos fármacos , Imunidade nas Mucosas/efeitos dos fármacos , Distribuição Aleatória
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