• Title/Summary/Keyword: Protein digestion

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Impact of Sodium Copper Chlorophyllin on Mercury Absorption Using an in Vitro Digestion with Human Intestinal Cell Model

  • Hwang, Han-Joon;Shim, Soon-Mi
    • Food Science and Biotechnology
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    • v.17 no.3
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    • pp.564-568
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    • 2008
  • The effects of sodium copper chlorophyllin (SCC) on bioaccessibility and uptake of mercury from fish were investigated using an in vitro digestion coupled with a Caco-2 cell. Fish along with SCC was subjected to a simulated in vitro digestion, which simulates both the gastric and small intestinal phase in vivo. Mercury bioaccessibility, the amount of mercury released from fish to aqueous phase following a digestion, was measured. Various amounts of SCC (0.1-25 mg) significantly reduced mercury bioaccessibility in a dose dependent manner by 49-89% compared to the negative control (fish without SCC) (p<0.05). Mercury bioaccessibility in varying molar ratios of mercury to positive control, 2,3-dimercapto-1-propane sulfonate (DMPS) was between 24 and 52%. Mercury uptake by Caco-2 cells from test media containing aqueous phase following in vitro digestion was measured after 6 hr incubation at $37^{\circ}C$. Cellular mercury uptake with increasing amount of SCC ranged from 0.352 to $0.052\;{\mu}g$ mercury/mg protein, while those in DMPS treatment were between 0.14 and $0.27\;{\mu}g$ mercury/mg protein. Our study suggests that SCC can reduce mercury absorption following fish consumption and may be efficient as a synthetic chelating agent for long term chronic mercury exposure in fish eating populations.

An Automated High Throughput Proteolysis and Desalting Platform for Quantitative Proteomic Analysis

  • Arul, Albert-Baskar;Han, Na-Young;Lee, Hookeun
    • Mass Spectrometry Letters
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    • v.4 no.2
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    • pp.25-29
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    • 2013
  • Proteomics for biomarker validation needs high throughput instrumentation to analyze huge set of clinical samples for quantitative and reproducible analysis at a minimum time without manual experimental errors. Sample preparation, a vital step in proteomics plays a major role in identification and quantification of proteins from biological samples. Tryptic digestion a major check point in sample preparation for mass spectrometry based proteomics needs to be more accurate with rapid processing time. The present study focuses on establishing a high throughput automated online system for proteolytic digestion and desalting of proteins from biological samples quantitatively and qualitatively in a reproducible manner. The present study compares online protein digestion and desalting of BSA with conventional off-line (in-solution) method and validated for real time sample for reproducibility. Proteins were identified using SEQUEST data base search engine and the data were quantified using IDEALQ software. The present study shows that the online system capable of handling high throughput samples in 96 well formats carries out protein digestion and peptide desalting efficiently in a reproducible and quantitative manner. Label free quantification showed clear increase of peptide quantities with increase in concentration with much linearity compared to off line method. Hence we would like to suggest that inclusion of this online system in proteomic pipeline will be effective in quantification of proteins in comparative proteomics were the quantification is really very crucial.

Enzymatic N-glycan analysis of 31 kDa molecule in plerocercoid of Spirometra mansoni (sparganum) and its antigenicity after chemical oxidation

  • Chung, Young-Bae;Kong, Yoon;Yang, Hyun-Jong
    • Parasites, Hosts and Diseases
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    • v.42 no.2
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    • pp.57-60
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    • 2004
  • A highly specific antigenic protein of 31 kDa from plerocercoid of Spirometra mansoni (sparganum) was obtained by gelatin affinity and Mono Q anion-exchange column chromatography. The purified 31 kDa protein was subjected to N-glycan enzymatic digestion for structural analysis. The relative electrophoretic mobility was analyzed by SDS-PAGE, before and after digestion. On SDS-PAGE after enzymatic digestion, the 31 kDa protein showed a molecular shift of approximately 2 kDa, which indicated the possession of complex N-linked oligosaccharides (N-glycosidase F sensitive) but not of high-mannose oligosaccharides (endo-beta-N-acetylglucosaminidase H, non-sensitive). Chemically periodated 31 kDa protein showed statistically non-significant changes with human sparganosis sera by enzyme linked immunosorbent assay (ELISA). Therefore, the dominant epitopes of the 31 kDa molecule in human sparganosis were found to be mainly polypeptide, while N-glycans of the antigenic molecule in sparganum was minimal in anti-carbohydrate antibody production.

A REVIEW OF THE MICROBIAL DIGESTION OF FEED PARTICLES IN THE RUMEN

  • McAllister, T.A.;Bae, H.D.;Yanke, L.J.;Cheng, K.J.;Ha, J.K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.7 no.3
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    • pp.303-316
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    • 1994
  • Microbial digestion of feed in the rumen involves a sequential attack culminating in the formation of fermentation products and microbial cells that can be utilized by the host animal. Most feeds are protected by a cuticular layer which is in effect a microbial barrier that must be penetrated or circumvented for digestion to proceed. Microorganisms gain access to digestible inner plant tissues through damage to the cuticle, or via natural cell openings (e.g., stomata) and commence digestion from within the feed particles. Primary colonizing bacteria adhere to specific substrates, divide to form sister cells and the resultant microcolonies release soluble substrates which attract additional microorganisms to the digestion site. These newly attracted microorganisms associate with primary colonizers to form complex multi-species consortia. Within the consortia, microorganisms combine their metabolic activities to produce the diversity of enzymes required to digest complex substrates (e.g., cellulose, starch, protein) which comprise plant tissues. Feed characteristics that inhibit the microbial processes of penetration, colonization and consortia formation can have a profound effect on the rate and extent of feed digestion in the rumen. Strategies such as feed processing or plant breeding which are aimed at manipulating feed digestion must be based on an understanding of these basic microbial processes and their concerted roles in feed digestion in the rumen.

Nutrition Value of Chlorella (II) (Chlorella Protein의 영양가(營養價)에 관한 연구(硏究) (II))

  • Park, Chung-Ung;Hwang, Ho-Kwan
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.3 no.1
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    • pp.43-46
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    • 1974
  • It was reported that the digestion ratio of chlorella was low because it had a low metabolic rate in body. Generally, the thickness of a cell membrane of it is $200-250\;{\AA}$, the weight of it is approximatly 13% of the total weight of a dry cell. And it is composed of protein, lipid, hemicellulose and ash etc. So, in order to elevate the digestion ratio of chlorella in body, we experimented the crude treatment methods of chlorella. The results obtained in this experiment are summarized as follows : 1. The digestion ratios calculated from ordinary N- balance method were 83.05% for 10% chl. (b) plus diastase group ; 81.25% for 10% chl. (b) plus amylase group, and 79.23% for 10% chl. (b), 58.55% for 10% chl. (a). 2. Biological values from this method were 80.25% for 10% chl. (b) plus diastase group, and 60% for 10% for chlorella(a).

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To develop the classification method of Agricultural by-productions for biogas production

  • Kim, Minjee;Kim, Sanghun
    • International Journal of Internet, Broadcasting and Communication
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    • v.7 no.2
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    • pp.155-160
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    • 2015
  • The objective of this study was to develop the classification method of various organic wastes. Specifically, the effects of proximate composition on the biogas production and degradation rates of agricultural by-production was investigated and a new standards for mixture of various organic wastes based on proximate composition combination was developed. Agricultural by-products (ABPs) with medium total carbohydrate, medium crude protein and low fat contents demonstrated the single step digestion process. ABPs with low total carbohydrate, high crude protein and high fat contents demonstrated the two step digestion process of Diauxic growth. The single ABP (Class No. 15) and the mixed ABPs (Class No. 12+18, 6+12+22, 9+12+18) after 10days showed the similar biogas yield pattern. We can use the classification method for the more ABPs and organic wastes from factory and municipal waste treatment plant for the high efficient biogas production.

Angiotensin I-converting Enzyme Inhibitory Activities of Porcine Skeletal Muscle Proteins Following Enzyme Digestion

  • Katayama, K.;Fuchu, H.;Sakata, A.;Kawahara, S.;Yamauchi, K.;Kawamura, Y.;Muguruma, M.
    • Asian-Australasian Journal of Animal Sciences
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    • v.16 no.3
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    • pp.417-424
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    • 2003
  • Inhibitory activities against angiotensin I-converting enzyme (ACE) of enzymatic hydrolysates of porcine skeletal muscle proteins were investigated. Myosin B, myosin, actin, tropomyosin, troponin and water-soluble proteins extracted from pork loin were digested by eight kinds of proteases, including pepsin, $\alpha$-chymotrypsin, and trypsin. After digestion, hydrolysates produced from all proteins showed ACE inhibitory activities, and the peptic hydrolysate showed the strongest activity. In the case of myosin B, the molar concentration of peptic hydrolysate required to inhibit 50% of the activity increased gradually as digestion proceeded. The hydrolysates produced by sequential digestion with pepsin and $\alpha$-chymotrypsin, pepsin and trypsin or pepsin and pancreatin showed weaker activities than those by pepsin alone, suggesting that ACE inhibitory peptides from peptic digestion might lose their active sequences after digestion by the second protease. However, the hydrolysates produced by sequential digestion showed stronger activities than those by $\alpha$-chymotrypsin, trypsin or pancreatin alone. These results suggested that the hydrolysates of porcine meat were able to show ACE inhibitory activity, even if they were digested in vivo, and that pork might be a useful source of physiologically functional factors.

Quality Control of Photosystem II during Photoinhibition

  • Yamamoto, Yasusi
    • Journal of Photoscience
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    • v.9 no.2
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    • pp.55-58
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    • 2002
  • The reaction center Dl protein of photosystem II is the target of photodamage by excess illumination. The Dl protein is damaged by reactive oxygen species generated by photochemical reactions and then degraded by specific proteolytic enzymes. We found that the Dl protein also cross-links with the surrounding polypeptides, such as D2 and CP43 in isolated thylakoids or photosystem II-enriched membranes from spinach under the illumination with strong visible light. The cross-linking was observed in spinach leaf discs as well when they were illuminated at higher temperature (40°C). It was also shown that the cross-linked products are digested efficiently by a protease(s) in the stroma. Thus the cross-linking/digestion processes of the Dl protein seem to comprise a new pathway in the turnover of the photodamaged Dl protein. It should be noted, however, that the cross-linked products of the Dl protein and CP43 induced by endogenous cationic radicals in the donor-side photoinhibition are resistant to proteolytic digestion. Accumulation of these cross-linked products in the thylakoids may lead to the decay of the function of chloroplasts and finally to the death of plant cells. Thus, we suggest that the quality control of photosystem II, especially removal of the cross-linked products of the Dl protein, is crucial for the survival of chloroplasts under the light stress.

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Effects of Micronization on the In situ and In vitro Digestion of Cereal Grains

  • McAllister, T.A.;Sultana, H.
    • Asian-Australasian Journal of Animal Sciences
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    • v.24 no.7
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    • pp.929-939
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    • 2011
  • The effects of micronization on in situ and in vitro nutrient disappearances of wheat, barley and corn were investigated in a series of experiments. In Experiment 1, chemical composition and in situ dry matter disappearance (DMD) of six varieties of wheat were determined. In addition, an in vitro study was completed using ground micronized and unmicronized wheat (var. Kansas). In Experiment 2, three varieties of wheat (Kansas, Sceptre and Laura) and in Experiment 3, three cereal grains (wheat, barley and corn) were either micronized for 1 min to attain internal kernel temperatures of 90-100$^{\circ}C$ or not (controls), and DM, protein and starch disappearances were estimated. In Experiment 2, an in vitro study was also completed using ground micronized and unmicronized wheat (var. Kansas). Wheat samples varied with respect to crude protein (10.0-21.2%), starch (61.6-73.9%), NDF (8.5-11.8%), volume weight (753-842 g/L) and kernel hardness (0.0-32.0). Rate (p = 0.003) and extent (p = 0.001) of in situ DMD differed among wheat varieties. Correlations between in situ kinetics, and chemical and physical properties of wheat varieties showed that protein content was negatively correlated with the rate of disappearance ($r^2$ = -0.77). Micronization of all grains markedly reduced (p = 0.001) the rate and extent of DM, and protein disappearances as compared to control samples. Micronization increased (p<0.05) the digestion of starch in wheat. However, release of ammonia into the incubation medium was markedly reduced (p<0.05), suggesting that micronization increased the resistance of protein to microbial digestion. Disappearances of DM, protein and starch differed (p = 0.001) among cereal grains with wheat>barley>corn. Micronization reduced the rate of DM disappearance (p = 0.011) and slowly degradable protein fractions (p = 0.03), however, increased (p = 0.004) slowly degradable starch fractions of all three cereals. Examination of in situ samples by scanning electron microscopy confirmed that microbial colonization focused on starch granules in micronized grains, and that the protein matrix exhibited resistance to microbial colonization. These results suggest that micronization may be used to increase the ruminal escape value of protein in cereal grains, but may lead to increased starch digestion if grains are finely ground.

Ex vivo Digestion of Milk from Red Chittagong Cattle Focusing Proteolysis and Lipolysis

  • Islam, Mohammad Ashiqul;Ekeberg, Dag;Rukke, Elling-Olav;Vegarud, Gerd Elisabeth
    • Asian-Australasian Journal of Animal Sciences
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    • v.28 no.4
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    • pp.559-567
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    • 2015
  • Ex vivo digestion of proteins and fat in Red Chittagong Cattle milk from Bangladesh was carried out using human gastrointestinal enzymes. This was done to investigate the protein digestion in this bovine breed's milk with an especial focus on the degradation of the allergenic milk proteins; ${\alpha}_{s1}$-casein and ${\beta}$-lactoglobulin and also to record the generation of peptides. Lipolysis of the milk fat and release of fatty acids were also under consideration. After 40 min of gastric digestion, all the ${\alpha}_s$-caseins were digested completely while ${\beta}$-lactoglobulin remained intact. During 120 min of duodenal digestion ${\beta}$-lactoglobulin was reduced, however, still some intact ${\beta}$-lactoglobulin was observed. The highest number of peptides was identified from ${\beta}$-casein and almost all the peptides from ${\kappa}$-casein and ${\beta}$-lactoglobulin were identified from the gastric and duodenal samples, respectively. No lipolysis was observed in the gastric phase of digestion. After 120 min of duodenal digestion, milk fat showed 48% lipolysis. Medium (C10:0 to C16:0) and long (${\geq}C17:0$) chain fatty acids showed 6% to 19% less lipolysis than the short (C6:0 to C8:0) chain fatty acids. Among the unsaturated fatty acids $C18:1{\sum}others$ showed highest lipolysis (81%) which was more than three times of $C18:2{\sum}all$ and all other unsaturated fatty acids showed lipolysis ranging from 32% to 38%. The overall digestion of Bangladeshi Red Cattle milk was more or less similar to the digestion of Nordic bovine milk (Norwegian Red Cattle).