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  1. Article: Evaluation of protein hydrolysis in raw sources by digestive proteases of Senegalese sole (Solea senegalensis, Kaup 1858) using a combination of an in vitro assay and sodium dodecyl sulphate polyacrylamide gel electrophoresis analysis of products

    Sáenz de Rodrigáñez, Miguel Ángel / Medina, Esther / Moyano, Francisco Javier / Alarcón, Francisco Javier

    Aquaculture research. 2011 Oct., v. 42, no. 11

    2011  

    Abstract: The hydrolysis of protein in different animal and plant sources by the intestinal proteases of Senegalese sole (Solea senegalensis) was studied using a combination of an in vitro digestibility assay and the evaluation of the protein fractionation by ... ...

    Abstract The hydrolysis of protein in different animal and plant sources by the intestinal proteases of Senegalese sole (Solea senegalensis) was studied using a combination of an in vitro digestibility assay and the evaluation of the protein fractionation by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS‐PAGE). In vitro hydrolysis was performed for 90 min in a closed reactor maintained at constant pH and temperature. Samples of the reaction mixture at different time intervals were developed in SDS‐PAGE gels to evaluate the progressive hydrolysis of the different protein fractions in each protein source. A numerical value [coefficient of protein degradation (CPD)], which integrates the information obtained after image analysis of the gels, is proposed for comparison among proteins, according to the intensity of the hydrolysis produced by sole proteases. Additionally, the total free amino acid released from proteins was measured during the in vitro assay. Casein, squid meal and soybean concentrate (SBC) proteins showed very similar degradation patterns, with a quick and almost complete proteolysis within the first minutes of the enzymatic reaction. Fish and krill meals were hydrolysed more progressively. On the contrary, pea meal (PM) and corn gluten meal (CGM) showed scarce changes in their protein profile after 90 min of reaction. For animal protein sources, the final CPD values ranged from 77.6% to 87.0%, showing not significant differences. By contrast, PM (30.5%) and CGM (32.3%) presented significantly lower CPD values (P<0.05) as compared with SBC (90.6%). In general, a linear fitting was found between CPD and the release of free amino acids during in vitro protein hydrolysis. The present study provides detailed information, which, combined with the conventional in vitro digestibility studies, may help in the evaluation of different raw sources according to their protein degradation patterns. This information can be applied directly to estimate the protein nutritional quality of ingredients for Senegalese sole feeds.
    Keywords Solea senegalensis ; animal source protein ; casein ; corn gluten meal ; enzymatic reactions ; feeds ; fish meal ; fractionation ; free amino acids ; gels ; hydrolysis ; image analysis ; in vitro digestibility ; in vitro studies ; ingredients ; nutritive value ; pH ; peas ; plants ; polyacrylamide gel electrophoresis ; protein degradation ; protein sources ; proteinases ; proteolysis ; sodium sulfate ; soybean meal ; squid ; temperature
    Language English
    Dates of publication 2011-10
    Size p. 1639-1652.
    Publishing place Blackwell Publishing Ltd
    Document type Article
    ZDB-ID 1227359-4
    ISSN 1355-557X
    ISSN 1355-557X
    DOI 10.1111/j.1365-2109.2010.02758.x
    Database NAL-Catalogue (AGRICOLA)

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  2. Article: Effect of diets containing a purified soybean trypsin inhibitor on growth performance, digestive proteases and intestinal histology in juvenile sea bream (Sparus aurata L.)

    Santigosa, Ester / Sáenz de Rodrigáñez, Miguel Ángel / Rodiles, Ana / Barroso, Fernando García / Alarcón, Francisco Javier

    Aquaculture research. 2010 Aug., v. 41, no. 9

    2010  

    Abstract: Juvenile sea bream were fed on diets containing 0.0, 2.0 or 4.0 g kg⁻¹ of a soybean trypsin inhibitor (SBTI) for 30 days. The growth performance, total protease activity and intestinal histology were studied after 0, 15 and 30 days of dietary treatment. ... ...

    Abstract Juvenile sea bream were fed on diets containing 0.0, 2.0 or 4.0 g kg⁻¹ of a soybean trypsin inhibitor (SBTI) for 30 days. The growth performance, total protease activity and intestinal histology were studied after 0, 15 and 30 days of dietary treatment. No significant differences were found in the weight gain, specific growth rate (SGR) and feed conversion rate in fish fed on inhibitor-supplemented diets when compared with those fed on an inhibitor-free diet. Only the SGR at day 15 decreased significantly with protease inhibitor inclusion, although this effect was not observed at day 30. In relation to proteolytic activity at day 15, the total protease activity in the distal intestine decreased in fish fed on inhibitor-supplemented diets. Zymograms of these extracts showed that the SBTI reduced the intensity of some proteolytic fractions in the distal intestine. A noticeable reduction in the protease activity of the intestinal content in fish fed on the highest level of soybean inhibitor (4.0 g kg⁻¹) was also observed. However, at day 30, the inhibition effect on these active bands was not detected, and the total protease activity was similar to that in fish fed on an inhibitor-free diet. Histological examination revealed no perceptible differences in the intestinal structure between any of the diet groups. In addition, all fish were maintained under experimentation for 10 more days and fed on an inhibitor-free diet to determine whether the possible effects caused by the protease inhibitor could be reverted. The administration of SBTI-supplemented diets did not affect sea bream growth performance or intestine histology after 30 days, and only a decrease in the total alkaline protease activity was found at day 15.
    Keywords Sparus aurata ; bream ; diet ; feed conversion ; feeding level ; fish feeding ; histology ; proteinases ; soybeans ; trypsin inhibitors ; weight gain
    Language English
    Dates of publication 2010-08
    Size p. e187-e198.
    Publisher Blackwell Publishing Ltd
    Publishing place Oxford, UK
    Document type Article
    ZDB-ID 1227359-4
    ISSN 1355-557X
    ISSN 1355-557X
    DOI 10.1111/j.1365-2109.2010.02500.x
    Database NAL-Catalogue (AGRICOLA)

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