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1.
Int J Biol Macromol ; 256(Pt 2): 128462, 2024 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-38042317

RESUMO

Chitin extraction from the shells of American lobsters (Homarus americanus) was optimized through the use of response surface methodology (RSM). The demineralization step was optimized to minimize the ash content of shell samples and the deproteination step was optimized to minimize the protein content of the chitin product. At a laboratory scale, one set of optimized conditions for the demineralization step was 7.35 % w/w acetic acid at a 40 mL/g of powdered lobster shell ratio for 15 min; this lowered the ash content from 39.62 % to 0.41 ± 0.08 %. A set of optimized conditions for the deproteination step at a similar scale was 4 % w/w sodium hydroxide at a 43 mL/g demineralized shell ratio heated to 95 °C for 83 min. These conditions were indicated to entirely remove protein from the resultant chitin. Average yields under optimized conditions were 23.43 ± 1.75 % for demineralization and 30.33 ± 0.02 % for deproteination, though a demineralization reaction with larger biomass input had a higher yield at 40.31 %.


Assuntos
Quitina , Decápodes , Animais , Quitina/química , Nephropidae , Decápodes/química , Exoesqueleto/química
2.
ACS Nano ; 13(4): 3796-3805, 2019 04 23.
Artigo em Inglês | MEDLINE | ID: mdl-30856331

RESUMO

Plastic packaging effectively protects foods from mechanical, microbial, and chemical damage, but oxygen can still permeate these plastics, degrading foods. Improving the gas barrier usually requires metallic or halogenated polymeric coatings; however, both cause environmental concerns and metallic coatings block visible light and electromagnetic signals. This paper reports a design of a highly flexible, visible light and radio frequency transparent coating on commercial poly(ethylene terephthalate) (PET) film. Nanoscale blending was achieved between negatively charged cellulose nanofibers and positively charged chitin nanowhiskers by employing spray-assisted layer-by-layer assembly. Synergetic interplay between these highly crystalline nanomaterials results in a flexible film with superior barrier characteristics. The oxygen transmission rate was below 0.5 mL m-2 day-1. Moreover, this coating maintains its performance even when exposed to common hazards such as bending stress and hydration. The coating also notably reduces the haziness of PET with a negligible loss of transparency and provides effective inhibition of antibacterial growth. This "crab-on-a-tree" nanocoating holds high potential for biorenewable and optical and radio frequency transparent packaging applications.


Assuntos
Celulose/química , Quitina/química , Embalagem de Alimentos , Nanoestruturas/química , Polietilenotereftalatos/química , Animais , Decápodes/química , Embalagem de Alimentos/métodos , Luz , Nanofibras/química , Oxigênio/química , Permeabilidade , Ondas de Rádio , Propriedades de Superfície , Madeira/química
3.
J AOAC Int ; 102(1): 278-285, 2019 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-30594270

RESUMO

Organotin compounds are anthropogenic metal species with multiple uses as pesticides, preservatives, antifouling agents, biocides, and catalysts. Butyltins are the main organotin compounds found in biota, and the highest levels are found in marine foodstuffs. In this paper, we present the figures of merit for an in-house validated method for routine analysis of butyltins in seafood using GC inductively coupled plasma isotope dilution MS. The working range of the method spanned several orders of magnitude from 3.3-1013, 2.4-785, and 0.3-900 ng Sn/g dry weight for monobutyltin (MBT), dibutyltin (DBT), and tributyltin (TBT), respectively. The trueness of the method was evaluated by analyzing Certified Reference Materials (CRMs) ERM CRM 477 (Mussel Tissue) and NIES CRM 15 (Scallop). Recoveries, with RSD % in parentheses, were 78 (±14), 80 (±6), and 88% (±8%) for MBT, DBT, and TBT in ERM CRM 477 and 96% (±5%) for TBT in NIES CRM 15. Good agreements were found between experimental uncertainties and uncertainties predicted for single-laboratory validated methods calculated from the maximum standard measurement uncertainty function. The method has proven to be robust, and the wide range of seafood validated ensures that the method is applicable for measuring butyltins in marine tissue.


Assuntos
Contaminação de Alimentos/análise , Cromatografia Gasosa-Espectrometria de Massas/métodos , Compostos Orgânicos de Estanho/análise , Alimentos Marinhos/análise , Compostos de Trialquitina/análise , Animais , Bivalves/química , Braquiúros/química , Decápodes/química , Gadiformes
4.
Curr Med Chem ; 25(19): 2245-2259, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-29110589

RESUMO

The alarming increase of antimicrobial resistance has led to a growing number of studies aiming to develop novel antimicrobial therapeutics. Natural antimicrobial peptides possess a potent and broad-spectrum antimicrobial activity combined with diverse and unique structural motifs, which confer their different mechanisms of action. These peptides are ubiquitous in organisms and are integral to the innate immune system. Recently, identification of antimicrobial peptides from marine crustaceans has become the centre of attention of many researchers. This increasing interest stems from the remarkable diversity in the structural and genetic composition of these peptides compared to terrestrial counterparts. Thus, peptides from marine crustaceans can serve as future templates for novel antimicrobial agents. Here, we provide an overview of various antimicrobial peptides from the marine crustaceans, their antimicrobial activity and structure- activity relationships. We also discuss the potential and challenges of their development as new antimicrobial agents.


Assuntos
Antibacterianos/farmacologia , Antifúngicos/farmacologia , Peptídeos Catiônicos Antimicrobianos/farmacologia , Sequência de Aminoácidos , Animais , Antibacterianos/química , Antibacterianos/isolamento & purificação , Antifúngicos/química , Antifúngicos/isolamento & purificação , Peptídeos Catiônicos Antimicrobianos/química , Peptídeos Catiônicos Antimicrobianos/isolamento & purificação , Decápodes/química , Descoberta de Drogas , Caranguejos Ferradura/química , Alinhamento de Sequência , Relação Estrutura-Atividade
5.
Glycoconj J ; 34(1): 85-94, 2017 02.
Artigo em Inglês | MEDLINE | ID: mdl-27658397

RESUMO

An N-acetyl sugar-binding lectin (termed iNoL) displaying cytotoxic activity against human cancer cells was isolated from the slipper lobster Ibacus novemdentatus (family Scyllaridae). iNoL recognized monosaccharides containing N-acetyl group, and glycoproteins (e.g., BSM) containing oligosaccharides with N-acetyl sugar. iNoL was composed of five subunits (330, 260, 200, 140, and 30 kDa), which in turn consisted of 70-, 40-, and 30-kDa polypeptides held together by disulfide bonds. Electron microscopic observations and gel permeation chromatography indicated that iNoL was a huge (500-kDa) molecule and had a polygonal structure under physiological conditions. iNoL displayed cytotoxic (apoptotic) effects against human cancer cell lines MCF7 and T47D (breast), HeLa (ovarian), and Caco2 (colonic), through incorporation (internalization) into cells. The lectin was transported into lysosomes via endosomes. Its cytotoxic effect and incorporation into cells were inhibited by the co-presence of N-acetyl-D-mannosamine (ManNAc). Treatment of HeLa cells with iNoL resulted in DNA fragmentation and chromatin condensation, through activation of caspase-9 and -3. In summary, the novel crustacean lectin iNoL is incorporated into mammalian cancer cells through glycoconjugate interaction, and has cytotoxic (apoptotic) effects.


Assuntos
Antineoplásicos/farmacologia , Apoptose/efeitos dos fármacos , Decápodes/química , Endocitose , Lectinas/farmacologia , Animais , Antineoplásicos/química , Células CACO-2 , Caspase 3/metabolismo , Caspase 9/metabolismo , Endossomos/efeitos dos fármacos , Endossomos/metabolismo , Glicoproteínas/metabolismo , Células HeLa , Humanos , Lectinas/química , Lectinas/toxicidade , Lisossomos/efeitos dos fármacos , Lisossomos/metabolismo , Células MCF-7 , Ligação Proteica
6.
Int J Biol Macromol ; 87: 163-71, 2016 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-26920243

RESUMO

Chitin was recovered through enzymatic deproteinization of the Norway lobster (Nephrops norvegicus) processing by-products. The obtained chitin was characterized and converted into chitosan by N-deacetylation, the acid-soluble form of chitin. Chitosan samples were then characterized by Fourier transform infrared spectroscopy (FTIR) and 13 Cross polarization magic angle spinning nuclear magnetic resonance (CP/MAS)-NMR spectroscopy. The antimicrobial activity and anti-proliferative capacity of chitosan were evaluated. Antimicrobial activity assays indicated that prepared chitosan exhibited marked inhibitory activity against the bacterial and fungal strains tested. Further, cytotoxic effects of chitosan samples on human colon carcinoma cells HCT116 was evaluated using the MTT assay. Chitosan showed the antiproliferative capacity against the colon-cancer-cell HCT116 in a dose dependent manner with IC50 of 4.6mg/ml. Indeed, HCT116 cell proliferation was significantly inhibited (p<0.05) between 13.5 and 67.5% at 0.5-6mg/mL of chitosan after 24h of cell treatment. The chitosan showed high antitumor activity which seemed to be dependent on its characteristics such as acetylation degree.


Assuntos
Antibacterianos/farmacologia , Antineoplásicos/farmacologia , Quitina/farmacologia , Quitosana/farmacologia , Decápodes/química , Resíduos Industriais , Acetilação , Animais , Antibacterianos/química , Antineoplásicos/química , Antineoplásicos/isolamento & purificação , Bactérias/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Quitina/química , Quitina/isolamento & purificação , Quitosana/química , Quitosana/isolamento & purificação , Células HCT116 , Humanos
7.
Appl Biochem Biotechnol ; 176(4): 1217-36, 2015 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-25935224

RESUMO

We studied the Cu(2+)-mediated inhibition and aggregation of Exopalaemon carinicauda arginine kinase (ECAK). We found that Cu(2+) significantly inactivated ECAK activity and double-reciprocal kinetics demonstrated that Cu(2+) induced noncompetitive inhibition of arginine and ATP (IC50 = 2.27 ± 0.16 µM; K i for arginine = 13.53 ± 3.76; K i for ATP = 4.02 ± 0.56). Spectrofluorometry results showed that Cu(2+) induced ECAK tertiary structural changes including the exposure of hydrophobic surfaces that directly induced ECAK aggregation. The addition of osmolytes such as glycine and proline successfully blocked ECAK aggregation induced by Cu(2+) and recovered ECAK activity. We built a 3D structure for ECAK using the ECAK ORF gene sequence. Molecular dynamics (MD) and docking simulations between ECAK and Cu(2+) were conducted to elucidate the binding mechanisms. The results showed that Cu(2+) blocked the entrance to the ATP active site; these results are consistent with the experimental result that Cu(2+) induced ECAK inactivation. Since arginine kinase (AK) plays an important role in cellular energy metabolism in invertebrates, our study can provide new information about the effect of Cu(2+) on ECAK enzymatic function and unfolding, including aggregation, and the protective effects of osmolytes on ECAK folding to better understand the role of the invertebrate ECAK metabolic enzyme in marine environments.


Assuntos
Arginina Quinase/antagonistas & inibidores , Proteínas de Artrópodes/antagonistas & inibidores , Cobre/química , Decápodes/química , Trifosfato de Adenosina/química , Sequência de Aminoácidos , Animais , Arginina/química , Arginina Quinase/química , Arginina Quinase/isolamento & purificação , Proteínas de Artrópodes/química , Proteínas de Artrópodes/isolamento & purificação , Domínio Catalítico , Cátions Bivalentes , Decápodes/enzimologia , Glicina/química , Interações Hidrofóbicas e Hidrofílicas , Cinética , Simulação de Acoplamento Molecular , Simulação de Dinâmica Molecular , Dados de Sequência Molecular , Prolina/química , Agregados Proteicos , Ligação Proteica , Dobramento de Proteína , Estrutura Secundária de Proteína
8.
Eur J Nutr ; 54(2): 301-7, 2015 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-24821271

RESUMO

AIM: This study investigated the hypoglycemic and antioxidant effects of shrimp astaxanthin on the kidney of alloxan-induced diabetic rats. METHODS: Animals were distributed into four groups of six rats each: a control group (C), a diabetic group (D), a diabetic group supplemented with Astaxanthin (D+As) dissolved in olive oil and a diabetic group supplemented with olive oil (D+OO). In vitro antidiabetic effect was tested in plasma and kidney tissue. RESULTS: The group D of rats showed significant (P < 0.05) increase of glycemia, creatinine, urea and uric acid levels compared to those of the control group (C). Moreover, plasma and kidney malondialdehyde (MDA) and protein carbonyl (PCO) levels for the rats of the group D were significantly increased compared to the control group. Contrariwise, antioxidant enzyme activities, such as catalase (EC 1.11.1.6), superoxide dismutase (EC 1.15.1.1) and non-enzymatic levels of reduced glutathione, were significantly (P < 0.05) decreased in the plasma and kidney of diabetic rats compared to the control ones. The astaxanthin supplementation in rats diet improved the antioxidant enzyme activities and significantly decreased the MDA and PCO levels compared to diabetic rats. Indeed, no significant (P ≥ 0.05) improvement was observed for the fourth group (D+OO) compared to the control group (C). Histological analysis of kidney showed glomerular hypertrophy and tubular dilatation for the diabetic rats. For D+As rats, these histopathological changes were less prominent. CONCLUSIONS: Our results suggest that shrimp astaxanthin may play an important role in reduction of oxidative damage and could prevent pathological changes in diabetic rats suggesting promising application of shrimp astaxanthin in diabet treatment.


Assuntos
Antioxidantes/uso terapêutico , Diabetes Mellitus Experimental/dietoterapia , Nefropatias Diabéticas/prevenção & controle , Suplementos Nutricionais , Rim/efeitos dos fármacos , Insuficiência Renal/prevenção & controle , Exoesqueleto/química , Animais , Antioxidantes/efeitos adversos , Antioxidantes/economia , Antioxidantes/metabolismo , Decápodes/química , Diabetes Mellitus Experimental/complicações , Diabetes Mellitus Experimental/metabolismo , Diabetes Mellitus Experimental/patologia , Nefropatias Diabéticas/fisiopatologia , Suplementos Nutricionais/efeitos adversos , Suplementos Nutricionais/economia , Indústria de Processamento de Alimentos/educação , Glutationa/antagonistas & inibidores , Glutationa/sangue , Glutationa/metabolismo , Hiperglicemia/prevenção & controle , Hipoglicemiantes/efeitos adversos , Hipoglicemiantes/economia , Hipoglicemiantes/metabolismo , Hipoglicemiantes/uso terapêutico , Rim/metabolismo , Rim/patologia , Rim/fisiopatologia , Peroxidação de Lipídeos , Masculino , Estresse Oxidativo , Oxirredutases/antagonistas & inibidores , Oxirredutases/sangue , Oxirredutases/metabolismo , Ratos Wistar , Insuficiência Renal/complicações , Insuficiência Renal/fisiopatologia , Resíduos/análise , Resíduos/economia , Xantofilas/efeitos adversos , Xantofilas/economia , Xantofilas/metabolismo , Xantofilas/uso terapêutico
9.
Mar Pollut Bull ; 86(1-2): 274-282, 2014 Sep 15.
Artigo em Inglês | MEDLINE | ID: mdl-25096583

RESUMO

Methods that quantify dissolved hydrocarbons are needed to link oil exposures to toxicity. Solid phase microextraction (SPME) fibers can serve this purpose. If fibers are equilibrated with oiled water, dissolved hydrocarbons partition to and are concentrated on the fiber. The absorbed concentration (Cpolymer) can be quantified by thermal desorption using GC/FID. Further, given that the site of toxic action is hypothesized as biota lipid and partitioning of hydrocarbons to lipid and fibers is well correlated, Cpolymer is hypothesized to be a surrogate for toxicity prediction. To test this method, toxicity data for physically and chemically dispersed oils were generated for shrimp, Americamysis bahia, and compared to test exposures characterized by Cpolymer. Results indicated that Cpolymer reliably predicted toxicity across oils and dispersions. To illustrate field application, SPME results are reported for oil spills at the Ohmsett facility. SPME fibers provide a practical tool to improve characterization of oil exposures and predict effects in future lab and field studies.


Assuntos
Exposição Ambiental , Hidrocarbonetos/análise , Hidrocarbonetos/toxicidade , Polímeros/química , Poluentes Químicos da Água/análise , Poluentes Químicos da Água/toxicidade , Animais , Cromatografia Gasosa , Decápodes/química , Decápodes/efeitos dos fármacos , Poluição por Petróleo , Microextração em Fase Sólida/instrumentação
10.
Anal Chim Acta ; 827: 40-6, 2014 May 27.
Artigo em Inglês | MEDLINE | ID: mdl-24832993

RESUMO

A new method of sample preparation was developed and is reported for the first time. The approach combines in-vial filtration with dispersive solid-phase extraction (d-SPE) in a fast and convenient cleanup of QuEChERS (quick, easy, cheap, effective, rugged, and safe) extracts. The method was applied to simultaneous analysis of 42 diverse pesticides and 17 environmental contaminants, including polycyclic aromatic hydrocarbons, polychlorinated biphenyls (PCBs), and flame retardants, in shrimp as the sample matrix. Final extracts were analyzed by both low-pressure gas chromatography - triple quadrupole tandem mass spectrometry (LPGC-MS/MS), and high-performance liquid chromatography - triple quadrupole tandem mass spectrometry (HPLC-MS/MS) to provide a wide scope of analysis for targeted analytes. During method development, several different commercial sorbents for d-SPE were investigated and compared with respect to analyte recoveries. The method was validated at 10, 50, and 100 ng g(-1) spiking levels (10-fold lower for PCBs), and the results for nearly all analytes were between 70 and 115% recoveries with ≤17% relative standard deviations. The method was shown to be simple, fast, and effective for multi-application analysis of chemical residues in the representative food and environmental marker matrix.


Assuntos
Decápodes/química , Filtração/métodos , Contaminação de Alimentos/análise , Resíduos de Praguicidas/análise , Resíduos de Praguicidas/isolamento & purificação , Frutos do Mar/análise , Extração em Fase Sólida/métodos , Métodos Analíticos de Preparação de Amostras , Animais , Poluentes Ambientais/análise , Poluentes Ambientais/isolamento & purificação
11.
Rev. biol. trop ; 61(1): 429-437, Mar. 2013. tab
Artigo em Espanhol | LILACS | ID: lil-674092

RESUMO

Nutritional value of seafood for human consumption is worldwide recognized. Some information have been generated in other countries, nevertheless, there is limited information describing the chemical composition of some fishery important species caught in the Gulf of Nicoya. For this reason, we studied the levels of proximal components of the edible parts (fresh) of three commercially important species. The meat samples of snook Centropomus unionesis, the shrimp Trachypenaeus byrdi and the bivalve Polymesoda radiata, were collected from the Puntarenas local fish market during the fishing season of February 2009 to January 2010. Proximate composition analysis was determined according to AOAC methodology, and evaluated the moisture content, and protein and lipid composition of shellfish meats. The results indicated that the moisture content ranged from 74.6-80.6g/100g for snook 76.9-80.0g/100g for shrimp and 77.9-89.5g/100g for green mussel. After the moisture, the protein was the most abundant chemical fraction (6.8 to 21g/100g) showing the highest values in February for the shrimp and green mussel, and December for snook. The largest fluctuations in the lipid content were found in the snook, ranging from 0.7g/100g to 5.6g/100g; the highest values in this fraction were found in shrimp, green mussel and snook, for July, February and April samples respectively. Considering these results, we concluded that fish and shrimp species studied are a good alternative for human consumption as a source of protein and low lipid content.


La información disponible sobre la composición química de algunos recursos pesqueros que se capturan en el Golfo de Nicoya es muy limitada. Por tal razón se determinaron los niveles de los componentes proximales de las partes comestibles (fresco) de tres especies de importancia comercial. Las muestras fueron adquiridas en el Mercado de Puntarenas entre febrero 2009 y enero 2001 durante la temporada de pesca. Los análisis de la composición proximal se determinaron según la metodología de AOAC. Los resultados indicaron que el contenido de humedad varió entre 74.6-80.6g/100g en robalo (Centropomus unionesis), 76.9-80.0g/100g en camarón (Trachypenaeus byrdi) y 77.9-89.5g/100g en almeja verde (Poymesoda radiata). Después de la humedad la fracción química más abundante fue la proteína presentando los valores más altos en diciembre para el robalo y febrero para el camarón y la almeja verde. Las mayores fluctuaciones en el contenido de lípidos se presentaron en el robalo, variando desde 0.7g/100g hasta 5.6g/100g. Los valores más altos en esta fracción fueron encontrados en julio, febrero y abril en camarones, almeja y robalo respectivamente. Se concluye que la especie de pescado y camarón estudiados son una buena alternativa para el consumo humano por ser una fuente importante de proteínas y por su bajo contenido en lípidos.


Assuntos
Animais , Humanos , Bivalves/química , Decápodes/química , Peixes , Valor Nutritivo , Frutos do Mar/análise , Costa Rica , Pesqueiros , Estações do Ano
12.
Environ Sci Technol ; 46(10): 5310-8, 2012 May 15.
Artigo em Inglês | MEDLINE | ID: mdl-22524970

RESUMO

Seafood samples from the fishing ground closure areas of Mississippi Gulf Coast that were affected by the Deepwater Horizon Oil Spill Disaster were collected and analyzed for twenty-five 2- to 6-ring PAHs, about one month after the first day of incident. A total of 278 seafood samples consisting of 86 fishes, 65 shrimps, 59 crabs, and 68 oysters were collected and analyzed weekly from May 27, 2010 until October 2010 and monthly thereafter until August 2011. Statistically higher levels of total PAHs were detected in all four types of seafood samples during early part of the sampling period compared to the later months. There was no significant concentration difference between PAHs detected in the oyster samples for the current study and the 10-year historical data from the NOAA Mussel Watch program. The PAH levels in the tested seafood samples were similar to those detected in commonly consumed processed foods purchased from local grocery stores and restaurants. Overall, the levels of PAHs in all the tested seafood samples collected within one-year period after the Oil Spill incident were far below the public health Levels of Concern (LOC) established jointly by the NOAA/FDA/Gulf Coast states under the protocol to reopen state and federal waters.


Assuntos
Monitoramento Ambiental , Poluição por Petróleo/análise , Hidrocarbonetos Policíclicos Aromáticos/análise , Alimentos Marinhos/análise , Animais , Bivalves/química , Braquiúros/química , Decápodes/química , Peixes , Manipulação de Alimentos , Geografia , Carne/análise , Mississippi , Ostreidae/química , Padrões de Referência , Restaurantes , Fatores de Tempo
13.
Adv Food Nutr Res ; 65: 73-105, 2012.
Artigo em Inglês | MEDLINE | ID: mdl-22361181

RESUMO

Marine organisms represent a valuable source of nutraceuticals and functional compounds. The biodiversity of the marine environment and the associated chemical diversity constitute a practically unlimited resource of novel active substances for the development of bioactive products. Recently, a great deal of interest has been expressed in marine-derived bioactive peptides because of their numerous beneficial health effects. Moreover, several studies have reported that marine bioactive peptides can be used as antihypertensive, antioxidative, anticoagulant, and antimicrobial components in functional foods or nutraceuticals and pharmaceuticals due to their therapeutic potential in the treatment or prevention of disease. In this chapter, we provide an overview of bioactive peptides derived from marine organisms as well as information about their biological properties and mechanisms of action with potential applications in different areas.


Assuntos
Organismos Aquáticos/química , Suplementos Nutricionais , Promoção da Saúde , Peptídeos/metabolismo , Animais , Decápodes/química , Suplementos Nutricionais/análise , Descoberta de Drogas , Equinodermos/química , Fermentação , Peixes/metabolismo , Alimentos em Conserva/análise , Alimentos em Conserva/microbiologia , Alimentos Especializados/análise , Humanos , Moluscos/química , Peptídeos/química , Peptídeos/farmacologia , Peptídeos/uso terapêutico , Conformação Proteica , Alimentos Marinhos/análise , Alimentos Marinhos/microbiologia , Frutos do Mar/análise , Frutos do Mar/microbiologia
14.
J Proteome Res ; 8(11): 5356-62, 2009 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-19891510

RESUMO

The identification of commercial shrimp species is a relevant issue to ensure correct labeling, maintain consumer confidence and enhance the knowledge of the captured species, benefiting both, fisheries and manufacturers. A proteomic approach, based on 2DE, tryptic in-gel digestion, MALDI-TOF MS, and ESI-MS/MS analyses, is proposed for the identification of shrimp species with commercial interest. MALDI-TOF peptide mass fingerprint from arginine kinase tryptic digests were used for the identification of seven commercial, closely related species of Decapoda shrimps. Further identification and characterization of these peptides was performed by CID on an ESI-IT instrument, database search and de novo sequence interpretation, paying special attention to differential, species-specific peptides. Fisheries and manufacturers may take advantage of this methodology as a tool for a rapid and effective seafood product identification and authentication, providing and guaranteeing the quality and safety of the foodstuffs to consumers.


Assuntos
Arginina Quinase/química , Decápodes/enzimologia , Peptídeos/análise , Espectrometria de Massas por Ionização por Electrospray/métodos , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/métodos , Animais , Arginina Quinase/genética , Arginina Quinase/metabolismo , Bases de Dados de Proteínas , Decápodes/química , Espectrometria de Massas , Dados de Sequência Molecular , Mapeamento de Peptídeos/instrumentação , Mapeamento de Peptídeos/métodos , Alinhamento de Sequência , Análise de Sequência de Proteína/métodos , Especificidade da Espécie , Espectrometria de Massas por Ionização por Electrospray/instrumentação , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/instrumentação
15.
Sci Total Environ ; 407(5): 1784-97, 2009 Feb 15.
Artigo em Inglês | MEDLINE | ID: mdl-19110296

RESUMO

Aquatic ecosystems of Doñana National Park (DNP) were monitored using Procambarus clarkii as bioindicator in four campaigns carried out between 2003 and 2004 to assess environmental quality possibly threatened by agrochemicals used in nearby areas. An integrated approach was carried out, by combining the responses of well-established biomarkers and the massive analysis of biological effects at the proteomic level. In sites potentially polluted, lower catalase, glucose-6-P dehydrogenase, and esterase activities, and higher malondialdehyde, metallothionein and glutathione levels were found. Two-dimensional gel electrophoresis resolved >2500 gill spots, and image analysis detected that 35 showed significant intensity differences between the reference site and the other seven sites studied. The superiority of proteomic approaches was clearly recognized in our study since four different protein expression patterns were established based in the fold-number of up-/down-regulation of the 35 differentially expressed proteins. Sites located within Doñana Biological Reserve were essentially free of contaminants and those near the DNP limits were only slightly polluted. The higher proteomic responses found at the upper "Rocina" and "Partido" courses indicate that non-persistent agrochemicals are mainly used in Doñana surroundings. The highest responses corresponded to rice growing areas placed between the Guadiamar stream and the Guadalquivir River, according to the extended and intensive use of agrochemicals in such areas.


Assuntos
Decápodes/química , Herbicidas/análise , Poluentes Químicos da Água/análise , Animais , Catalase/análise , Decápodes/enzimologia , Decápodes/metabolismo , Eletroforese em Gel Bidimensional , Esterases/análise , Brânquias/química , Brânquias/metabolismo , Glucosefosfato Desidrogenase/análise , Glutationa/análise , Herbicidas/metabolismo , Malondialdeído/análise , Metalotioneína/análise , Portugal , Proteômica/métodos , Poluentes Químicos da Água/metabolismo
16.
Appl Microbiol Biotechnol ; 79(5): 785-91, 2008 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-18521593

RESUMO

Acetes chinensis is an underutilized shrimp species thriving in Bo Hai Gulf of China. Its hydrolysate digested with protease SM98011 has been previously shown to have high angiotensin I-converting enzyme (ACE) inhibitory activity (He et al., J Pept Sci 12:726-733, 2006). In this article, A. chinensis were fermented by Lactobacillus fermentum SM 605 and the fermented sauce presented high ACE inhibitory activity. The minimum IC(50) value (3.37 +/- 0.04 mg/mL) was achieved by response surface methodology with optimized process parameters such as fermentation time of 24.19 h, incubation temperature at 38.10 degrees C, and pH 6.12. Three ACE inhibitory peptides are purified by ultrafiltration, gel filtration, and reverse-phase high performance liquid chromatography. Identified by mass spectrometry, their amino acid sequences are Asp-Pro, Gly-Thr-Gly, and Ser-Thr, with IC(50) values of 2.15 +/- 0.02, 5.54 +/- 0.09, and 4.03 +/- 0.10 microM, respectively. Also, they are all novel ACE inhibitory peptides. Compared with protease digestion, fermentation is a simpler and cheaper method to produce ACE inhibitory peptides from shrimp A. chinensis.


Assuntos
Inibidores da Enzima Conversora de Angiotensina/isolamento & purificação , Decápodes/metabolismo , Fermentação , Limosilactobacillus fermentum/metabolismo , Peptídeos/isolamento & purificação , Inibidores da Enzima Conversora de Angiotensina/química , Inibidores da Enzima Conversora de Angiotensina/metabolismo , Inibidores da Enzima Conversora de Angiotensina/farmacologia , Animais , Decápodes/química , Peptídeos/química , Peptídeos/metabolismo , Peptídeos/farmacologia , Peptidil Dipeptidase A/metabolismo
17.
J Proteome Res ; 7(1): 424-31, 2008 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-18072731

RESUMO

Rab GTPases are involved in phagosome formation and maturation. However, the role of Rab GTPases in phagocytosis against virus infection remains unknown. In this study, it was found that a Rab gene ( PjRab) from marine shrimp was upregulated in virus-resistant shrimp, suggesting that Rab GTPase was involved in the innate response to virus. The RNAi and mRNA assays revealed that the PjRab protein could regulate shrimp hemocytic phagocytosis through a protein complex consisting of the PjRab, beta-actin, tropomyosin, and envelope protein VP466 of shrimp white spot syndrome virus (WSSV). It was further demonstrated that the PjRab gene silencing by RNAi caused the increase in the number of WSSV copies, indicating that the PjRab might be an intracellular virus recognition protein employed by a host to increase the phagocytic activity. Therefore, our study presents a novel Rab-dependent signaling complex, in which the Rab GTPase might detect virus infection as an intracellular virus recognition protein and trigger downstream phagocytic defense against virus in crustacean for the first time. This discovery would improve our understanding of the still poorly understood molecular events involved in innate immune response against virus infection of invertebrates.


Assuntos
Decápodes/imunologia , Complexos Multiproteicos/imunologia , Fagocitose/imunologia , Proteínas rab de Ligação ao GTP/fisiologia , Actinas , Animais , Decápodes/química , Tropomiosina , Regulação para Cima , Proteínas do Envelope Viral , Viroses/imunologia , Vírus da Síndrome da Mancha Branca 1/imunologia
18.
Artigo em Chinês | MEDLINE | ID: mdl-17357463

RESUMO

OBJECTIVE: To prepare carboxymethylchitin and study its properties. METHODS: Chitin was prepared from fresh shrimp shells and then carboxymethylchitin was prepared by the methods of alkalization and etherification as well as by the purification technique. The deacetylation degree of carboxymethylchitin was determined by the double-jump potentiometric titration method; the substitution degree was determined by the element analysis method; the carboxymethyl substitution position was analyzed by the Fourier transform infrared spectroscopy apparatus and the nuclear magnetic resonance spectroscopy apparatus; the relative molecular weight and its polydispersity were determined by the gel permeation chromatography with the multiple angle laser light scattering detection; the biological properties were tested according to the GB/T 16886 biological evaluation on medical devices. RESULTS: Carboxymethylchitin could be prepared by alkalization and etherification from chitin which was prepared from fresh shrimp shells by decalcification and deproteinization. The deacetylation degree of carboxymethylchitin was 13.76% according to the double-jump potentiometric titration; the degrees of deacetylation and substitution were 14.53% and 1.2390 respectively according to the element analysis. The IR spectrum showed that the substitutive position was N, O-substitution, and the 13C-NMR spectrum showed that substitutive position of carboxymethylchitin was mostly primary substitution of 6-OH, and according to the substitutive proportion, the substitutive turns were in the following decreasing order: 6-OH, NH2, and 3-OH. The weight-averaged and the number-averaged molecular weights and polydispersity were 6. 25 x 10(5), 5.60 x 10(5) and 1.22, respectively. The results from the biological property test showed that carboxymethylchitin was a biomaterial that was sterile, pyrogen-free, acute toxicity-free, cytotoxicity-free, intracutaneous irritation-free, skin sensitization-free and biomaterial genotoxicity-free, with no side or adverse effects on the related tissues after implantation into the human body. CONCLUSION: Carboxymethylchitin prepared from chitin by alkalization and etherification is a macromolecule biomaterial that has a low degree of deacetylation, a high degree of substitution, and a good biocompatibility.


Assuntos
Materiais Biocompatíveis , Quitina/análogos & derivados , Decápodes/química , Animais , Quitina/química , Quitina/toxicidade , Teste de Materiais , Camundongos , Estrutura Molecular , Peso Molecular , Espectroscopia de Infravermelho com Transformada de Fourier , Testes de Toxicidade
19.
Bioresour Technol ; 98(18): 3499-505, 2007 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-17317156

RESUMO

Twelve kinds of marine protein materials, including fish, shrimp, seashell, algae and seafood wastes were selected for the hydrolysis using four different proteases. The IC(50) values for angiotensin-converting enzyme (ACE) inhibitory activity of 48 hydrolysates were rapidly determined by capillary electrophoresis (CE). The values ranged from 0.17 to 501.7mg/ml, and were affected by both the marine protein resources and the selected proteases. Hydrolysates of the lowest IC(50) values were from shrimp (Acetes chinensis), shark meat, mackerel bone, Polysiphonia urceolata and Spirulina platensis, indicating these five kinds of marine food proteins contained beneficial materials for the production of ACE inhibitory peptides by proteolysis. The hydrolysates obtained using proteases Protamex and SM98011 had lower IC(50) values, showing these two proteases were superior to others. The CE method achieved the same sensitivity as the high performance liquid chromatography (HPLC) method. However, the CE method was faster and, as a result, more economical. Therefore, CE had potential for rapid screening of marine protein hydrolysates enriched in ACE inhibitory peptides.


Assuntos
Inibidores da Enzima Conversora de Angiotensina/química , Inibidores da Enzima Conversora de Angiotensina/farmacologia , Peptídeos/química , Peptídeos/farmacologia , Hidrolisados de Proteína/química , Hidrolisados de Proteína/farmacologia , Animais , Osso e Ossos/química , Cianobactérias/química , Decápodes/química , Eucariotos/química , Peixes , Carne , Pele/química
20.
Chemosphere ; 67(9): S36-47, 2007 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-17223177

RESUMO

The contamination and distribution of organochlorinated compounds were considered in three crustacean species (edible crab, Cancer pagurus; spider crab, Maja brachydactyla; velvet swimming crab, Necora puber) from five sites along the coasts of Brittany and Normandy (Western and North-Western France). PCBs (16 single congeners), pp'-DDE and HCB were measured in hepatopancreas, gonads and muscle: in all, 175 samples were analysed. The spider crab was the only species found in the five sampling sites, thus enabling comparison between areas. Specimens from Antifer were much more contaminated (summation operator 16 PCBs in hepatopancreas=2000-4000 ng g(-1) dry weight) than those from other sites (50-1000 ng g(-1) d.w.). Among all the three species, the spider crab appeared more contaminated by PCBs than the edible crab, by a factor 2-3, probably in relation with specific differences in their life cycle. There was no difference due to the gender of the species. Within the different analysed tissues, contamination levels increased from muscle to gonads and hepatopancreas in relation with the fat content. A very similar PCB composition was observed in all samples, PCB fingerprints being characterised by the relative importance of the more persistent PCB congeners: CB153, 138, 180, 187, and 118. Finally, these results were compared to recent food regulations first of maximum marker PCB intake and secondly of maximum dioxin-like PCB intake. By considering the muscle, all samples were far below the regulatory limits; for hepatopancreas and gonads, however, some samples were unfit for human consumption.


Assuntos
Decápodes/química , Contaminação de Alimentos/análise , Hidrocarbonetos Clorados/análise , Poluentes Químicos da Água/análise , Animais , França , Geografia , Gônadas/química , Hepatopâncreas/química , Humanos , Hidrocarbonetos Clorados/metabolismo , Carne/normas , Músculos/química , Bifenilos Policlorados/análise , Bifenilos Policlorados/metabolismo , Fatores de Risco , Poluentes Químicos da Água/metabolismo
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