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1.
Appl Biochem Biotechnol ; 180(5): 1016-1027, 2016 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-27287996

RESUMO

Sugar beet pulp (SBP) and molasses, as an agro industrial waste material, are produced in large amounts annually. Thus, a major challenge nowadays is to develop procedures that could increase the value of the generated waste. In this study, SBP as a support for cell immobilization and molasses as a source of nutrients were used for a dextransucrase (DS) production by Leuconostoc mesenteroides T3. The influence of SBP in native form (SBP-N) and after treatment with NaOH (SBP-NaOH) on DS production was investigated. The optimal medium composition for the maximum DS production was determined by varying the concentration of molasses, SBP, and sucrose. The maximum DS yield of 2.02 U/ml was obtained in the medium with 2.5 % of molasses, 2.5 % SBP-NaOH, and 4 % of sucrose concentration. Scanning electron microscopy (SEM) showed immobilization of Lc. mesenteroides T3 cells onto SBP-NaOH. According to the obtained results, the production of DS on molasses could be improved by using NaOH-treated SBP as a carrier for whole-cell immobilization. Our study reveals the basis for the development of process for DS production with additional reduction of expenses by using waste materials for obtaining the valuable biotechnological product.


Assuntos
Beta vulgaris/metabolismo , Glucosiltransferases/biossíntese , Leuconostoc mesenteroides/metabolismo , Melaço/análise , Técnicas de Cultura Celular por Lotes , Beta vulgaris/efeitos dos fármacos , Beta vulgaris/ultraestrutura , Células Imobilizadas/metabolismo , Fermentação/efeitos dos fármacos , Leuconostoc mesenteroides/efeitos dos fármacos , Sacarose/farmacologia
2.
Biotechnol Prog ; 32(4): 1046-55, 2016 07 08.
Artigo em Inglês | MEDLINE | ID: mdl-27254304

RESUMO

The present study investigated preparation of bovine and porcine erythrocyte membranes from slaughterhouse blood as bio-derived materials for delivery of dexamethasone-sodium phosphate (DexP). The obtained biomembranes, i.e., ghosts were characterized in vitro in terms of morphological properties, loading parameters, and release behavior. For the last two, an UHPLC/-HESI-MS/MS based analytical procedure for absolute drug identification and quantification was developed. The results revealed that loading of DexP into both type of ghosts was directly proportional to the increase of drug concentration in the incubation medium, while incubation at 37°C had statistically significant effect on loaded amount of DexP (P < 0.05). The encapsulation efficiency was about fivefold higher in porcine compared to bovine ghosts. Insight into ghosts' surface morphology by field emission-scanning electron microscopy and atomic force microscopy confirmed that besides inevitable effects of osmosis, DexP inclusion itself had no observable additional effect on the morphology of the ghosts carriers. DexP release profiles were dependent on erythrocyte ghost type and amount of residual hemoglobin. However, sustained DexP release was achieved and shown over 3 days from porcine ghosts and 5 days from bovine erythrocyte ghosts. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:1046-1055, 2016.


Assuntos
Dexametasona/análogos & derivados , Membrana Eritrocítica/metabolismo , Eritrócitos/metabolismo , Animais , Bovinos , Cromatografia Líquida de Alta Pressão , Dexametasona/química , Dexametasona/metabolismo , Membrana Eritrocítica/química , Eritrócitos/química , Suínos , Espectrometria de Massas em Tandem , Fatores de Tempo
3.
Colloids Surf B Biointerfaces ; 122: 250-259, 2014 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-25051307

RESUMO

The present study was aimed at investigating the effect of isolation process-gradual hypotonic hemolysis on chosen parameters of the erythrocyte membranes (ghosts) originating from bovine and porcine slaughterhouse blood. The estimation of the gradual hypotonic hemolysis as a drug loading procedure for the erythrocyte ghosts was performed as well. Based on the results derived from analysis of the osmotic properties of the erythrocytes, the gradual hemolysis was performed with high volume of erythrocytes and 35mM hypotonic sodium-phosphate/NaCl, enabling >90% of hemolysis for both types of erythrocytes. Detailed insight into ghosts' morphology by field emission-scanning electron microscopy revealed a distortion from erythrocyte shape and an altered surface texture with increased bilayer curvature for both samples. Compared to erythrocytes, an average diameter of ghosts from both type of erythrocytes decreased for only about 10%. The reported unidispersity of the isolated ghosts is of great importance for their potential application as vehicles of active compounds. Gradual hemolysis did not lead to substantial loss of cholesterol and membrane/cytoskeleton proteins. This result indicated the ghosts' possibility to mimic the chemical and structural anisotropic environment of in vivo cell membranes, which is of significance for drug diffusion and partition coefficients. Induced shift of phosphatidylserine to external surface of the ghosts demonstrated their potential application as vehicles for targeted drug delivery to cells of reticuloendothelial system. Ultra high-performance liquid chromatography and Fourier transform infrared spectroscopy revealed the presence of a drug model - dexamethasone-sodium phosphate, and its interaction with structural components in both types of erythrocyte ghosts.


Assuntos
Matadouros , Sangue , Membrana Eritrocítica , Hemólise/efeitos dos fármacos , Animais , Bovinos , Eletroforese em Gel de Poliacrilamida , Microscopia Eletrônica de Varredura , Espectroscopia de Infravermelho com Transformada de Fourier , Suínos
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