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2.
Front Bioeng Biotechnol ; 9: 725578, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-34527660

RESUMEN

Production of caproic and caprylic acid through anaerobic fermentation of crops or residual and waste biomass has been regarded as an alternative to the conventional ways, where plant oils and animal fats are mostly used. The downstream processing of the fermentation broth is a particular challenge since the broth has a highly complex composition and low concentrations of the target products. In this study, the proof-of-principle for a separation cascade for caproic (C6) and caprylic acid (C8) produced in a maize silage-based fermentation process was demonstrated. For clarification of the fermentation broth, a filter press and a ceramic ultrafiltration membrane was used to remove coarse solids and to separate suspended particles and macromolecules from the fermentation broth, respectively. With both techniques, the dry matter content was reduced from 6.8 to 2.3% and a particle-free product solution was obtained. Subsequently, the carboxylic acids were extracted with oleyl alcohol by liquid-liquid extraction with an extraction efficiency of 85% for C6 and 97% for C8. Over the whole cascade, 58% of caproic acid and 66% of caprylic acid were recovered from the fermentation broth into the extract. Among all separation steps, solid-liquid separation with the filter press caused the major part of the product loss of 21% of each carboxylic acid. By using separation equipment with a better solid separation efficiency such as decanter centrifuges or belt filter presses this loss could be minimized.

3.
Naunyn Schmiedebergs Arch Pharmacol ; 394(5): 941-954, 2021 05.
Artículo en Inglés | MEDLINE | ID: mdl-33284399

RESUMEN

Binary enterotoxins Clostridioides difficile CDT toxin, Clostridium botulinum C2 toxin, and Clostridium perfringens iota toxin consist of two separate protein components. The B-components facilitate receptor-mediated uptake into mammalian cells and form pores into endosomal membranes through which the enzymatic active A-components translocate into the cytosol. Here, the A-components ADP-ribosylate G-actin which leads to F-actin depolymerization followed by rounding of cells which causes clinical symptoms. The protein folding helper enzymes Hsp90, Hsp70, and peptidyl-prolyl cis/trans isomerases of the cyclophilin (Cyp) and FK506 binding protein (FKBP) families are required for translocation of A-components of CDT, C2, and iota toxins from endosomes to the cytosol. Here, we demonstrated that simultaneous inhibition of these folding helpers by specific pharmacological inhibitors protects mammalian, including human, cells from intoxication with CDT, C2, and iota toxins, and that the inhibitor combination displayed an enhanced effect compared to application of the individual inhibitors. Moreover, combination of inhibitors allowed a concentration reduction of the individual compounds as well as decreasing of the incubation time with inhibitors to achieve a protective effect. These results potentially have implications for possible future therapeutic applications to relieve clinical symptoms caused by bacterial toxins that depend on Hsp90, Hsp70, Cyps, and FKBPs for their membrane translocation into the cytosol of target cells.


Asunto(s)
ADP Ribosa Transferasas/toxicidad , Toxinas Bacterianas/toxicidad , Toxinas Botulínicas/toxicidad , Enterotoxinas/toxicidad , Animales , Células CACO-2 , Chlorocebus aethiops , Ciclofilinas/antagonistas & inhibidores , Ciclofilinas/metabolismo , Proteínas HSP70 de Choque Térmico/antagonistas & inhibidores , Proteínas HSP70 de Choque Térmico/metabolismo , Proteínas HSP90 de Choque Térmico/antagonistas & inhibidores , Proteínas HSP90 de Choque Térmico/metabolismo , Humanos , Proteínas de Unión a Tacrolimus/antagonistas & inhibidores , Proteínas de Unión a Tacrolimus/metabolismo , Células Vero
4.
J Anim Sci Biotechnol ; 11: 20, 2020.
Artículo en Inglés | MEDLINE | ID: mdl-32158542

RESUMEN

BACKGROUND: The hypothesis was tested that insect meal (IM) as protein source influences intermediary metabolism of growing pigs. To test this, 30 male, 5-week-old crossbred pigs were randomly assigned to 3 groups of 10 pigs each with similar body weights (BW) and fed isonitrogenous diets either without (CON) or with 5% IM (IM5) or 10% IM (IM10) from Tenebrio molitor L. for 4 weeks and key metabolic tissues (liver, muscle, plasma) were analyzed using omics-techniques. RESULTS:  Most performance parameters did not differ across the groups, whereas ileal digestibilities of most amino acids were 6.7 to 15.6%-units lower in IM10 than in CON (P < 0.05). Transcriptomics of liver and skeletal muscle revealed a total of 166 and 198, respectively, transcripts differentially expressed between IM10 and CON (P < 0.05). Plasma metabolomics revealed higher concentrations of alanine, citrulline, glutamate, proline, serine, tyrosine and valine and a lower concentration of asparagine in IM10 than in CON (P < 0.05). Only one out of fourteen quantifiable amino acid metabolites, namely methionine sulfoxide (MetS), in plasma was elevated by 45% and 71% in IM5 and IM10, respectively, compared to CON (P < 0.05). Plasma concentrations of both, major carnitine/acylcarnitine species and bile acids were not different across groups. Lipidomics of liver and plasma demonstrated no differences in the concentrations of triacylglycerols, cholesterol and the main phospholipids, lysophospholipids and sphingolipids between groups. The percentages of all individual phosphatidylcholine (PC) and phosphatidylethanolamine (PE) species in the liver showed no differences between groups, except those with 6 double bonds (PC 38:6, PC 40:6, PE 38:6, PE 40:6), which were markedly lower in IM10 than in CON (P < 0.05). In line with this, the percentage of C22:6n-3 in hepatic total lipids was lower in IM10 than in the other groups (P < 0.05). CONCLUSIONS: Comprehensive analyzes of the transcriptome, lipidome and metabolome of key metabolic tissues indicate that partial or complete replacement of a conventional protein source by IM in the diet has only a weak impact on the intermediary metabolism of growing pigs. Thus, it is concluded that IM from Tenebrio molitor L. can be used as a dietary source of protein in pigs without causing adverse effects on metabolism.

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