• Title/Summary/Keyword: Protein Digestion

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The Role of Milk Products in Metabolic Health and Weight Management

  • Zemel, Michael B.
    • Journal of Dairy Science and Biotechnology
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    • v.28 no.1
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    • pp.17-28
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    • 2010
  • A substantial body of evidence has emerged over the last decade in support of the novel concept that dietary calcium and dairy foods play an important role in regulating energy metabolism and thereby promote healthy weight management and reduce obesity risk. This concept has been demonstrated in experimental animals studies, cross-sectional and prospective population studies and a number of randomized clinical trials. Notably, the effects of dairy foods in weight management are more consistent than the effects of supplemental calcium across clinical trials, and calcium per se is responsible for approximately 40-50% of the effects of dairy. The calcium component is only effective in individuals with chronically low calcium intake, as it serves to prevent the endocrine response to low calcium diets which otherwise favors adipocyte energy storage; calcium also serves to promote energy loss via formation of calcium soaps in the gastrointestinal tract and thereby reduce fat absorption. The calcium-independent anti-obesity bioactivity of dairy resides primarily in whey. The key components identified to date are leucine and bioactive peptides resulting from whey protein digestion. The high concentration of leucine in whey stimulates a repartitioning of dietary energy from adipose tissue to skeletal muscle where it provides the energy required for leucine-stimulated protein synthesis, resulting in increased loss of adipose tissue and preservation of skeletal muscle mass during weight loss. Finally, dairy rich diets suppress the oxidative and inflammatory responses to obesity and thereby attenuate the diabetes and cardiovascular disease risk associated with obesity.

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Design and Expression of Recombinant Antihypertensive Peptide Multimer Gene in Escherichia coli BL21

  • Rao, Shengqi;Su, Yujie;Li, Junhua;Xu, Zhenzhen;Yang, Yanjun
    • Journal of Microbiology and Biotechnology
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    • v.19 no.12
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    • pp.1620-1627
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    • 2009
  • The design and expression of an antihypertensive peptide multimer (AHPM), a common precursor of 11 kinds of antihypertensive peptides (AHPs) tandemly linked up according to the restriction sites of gastrointestinal proteases, were explored. The DNA fragment encoding the AHPM was chemically synthesized and cloned into expression vector pGEX-3X. After an optimum induction with IPTG, the recombinant AHPM fused with glutathione S-transferase (GST-AHPM) was expressed mostly as inclusion body in Escherichia coli BL21 and reached the maximal production, 35% of total intracellular protein. The inclusion body was washed, dissolved, and purified by cation-exchange chromatography under denaturing conditions, followed by refolding together with size-exclusion chromatography and gradual dialysis. The resulting yield of the soluble GSTAHPM (34 kDa) with a purity of 95% reached 399 mg/l culture. The release of high active fragments from the AHPM was confirmed by the simulated gastrointestinal digestion. The results suggest that the design strategy and production method of the AHPM will be useful to obtain a large quantity of recombinant AHPs at a low cost.

Characterization of a Collagenase-1 Inhibitory Peptide Purified from Skate Dipturus chilensis Skin (홍어류(Dipturus chilensis) 껍질로부터 분리 정제된 collagenase-1 저해 펩타이드의 특성)

  • Park, Sung-Ha;Lee, Jung-Kwon;Jeon, Joong-Kyun;Byun, Hee-Guk
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.44 no.5
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    • pp.456-463
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    • 2011
  • We attempted to isolate a collagenase-1 inhibitory peptide from skate Dipturus chilensis skin protein. The protein from skate skin was digested by various enzymes (alcalase, ${\alpha}$-chymotrypsin, neutrase, papain, pepsin, and trypsin) to produce a collagenase-1 inhibitory peptide. The collagenase-1 inhibitory activity of the peptides obtained was measured by gelatin digestion assay. Among the six hydrolysates, pepsin hydrolysate exhibited the highest collagenase-1 inhibitory activity. The peptide showing strong collagenase-1 inhibitory activity was purified by Sephadex G-25 gel chromatography and HPLC using an octadecylsilyls (ODS) column. The amino acid sequence of purified collagenase-1 inhibitory peptide was identified to be Asn-Leu-Asp-Val -Leu-Glu-Val-Phe (961 Da) by quadrupole time of flight (Q-TOF) and electrospray ionization mass spectrometry (ESI-MS) mass spectroscopy. The $IC_{50}$ value of purified peptide was 87.0 ${\mu}M$. Moreover, the peptide did not exhibit cytotoxic effects on human dermal fibroblast cell lines.

Rapid Isolation of Genomic DNA from Normal and Apoptotic Cells Using Magnetic Silica Resins

  • Park, Jee-Sun;Park, Jung-Hyun;Na, Shin-Young;Choe, Soo-Young;Choi, Sang-Nam;You, Kwan-Hee
    • Journal of Microbiology and Biotechnology
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    • v.11 no.5
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    • pp.890-894
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    • 2001
  • The isolation of genomic DNA from mammalian cells is usually performed by cell lysis followed by protein digestion, extraction, and finally, ethanol precipitation of the chromosomal DNA. However, in the case of large sample numbers or when only small amounts of starting materials are available, such conventional methods are not efficient and are cumbersome to be applied. Some alternative methods have been described as well as having commercial DNA isolation kits to be available, nevertheless, there is room left for much improvement. In the present study, a novel method is introduced, where it simplifies conventional protocols by omitting some time-consuming steps such as protease incubation or DNA precipitation and its resuspension. Using paramagnetic silica resins, the genomic DNA was purified over a magnetic field, and the bound DNA was eluted with a low-salt buffer. The fidelity and effectiveness of this novel method was determined by using normal and apoptotic cells as a starting material and then compared to other protocols. The high speed and convenience along with its high efficiency in detecting apoptotic chromosomal DNA will prove this method to be an improved alternative in the isolation of genomic DNA from mammalian cells.

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Relative Palatability to Sheep of Some Browse Species, their In sacco Degradability and In vitro Gas Production Characteristics

  • Abdulrazak, S.A.;Nyangaga, J.;Fujihara, T.
    • Asian-Australasian Journal of Animal Sciences
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    • v.14 no.11
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    • pp.1580-1584
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    • 2001
  • A study was conducted to estimate the nutritive value of some selected acacia forages using palatability index, in sacco degradability and in vitro gas production characteristics. Ten wethers (mean wt. $18{\pm}3.5kg$) were offered Acacia tortilis, Acacia nilotica, Acacia mellifera, Acacia brevispica, Acacia Senegal and Leucaena leucocephala (control) using a cafeteria system to determine the species preference by the animals. The acacia species were rich in nitrogen and showed variable palatability pattern. Significant (p<0.05) differences in relative palatability index (RPI) were detected among the species with the following ranking: brevispica > leucaena > mellifera > tortilis > Senegal > nilotica. Acacia nilotica appeared to be of low relative palatability with RPI of 24% and this was attributed to relatively high phenolic concentrations. The DM potential degradability (B) and rate of degradation (c) of the species were significantly (p<0.05) different, ranging from 40.1 to 59.1% and 0.0285 to 0.0794/h respectively. Acacia species had moderate levels of rumen undegradable protein, much higher than that in leucaena. In vitro gas production results indicated the effect of polyphenolic compounds on the fermentation rate, with lower gas production recorded from A. nilotica and tortilis. Based on RPI, A. brevispica and mellifera were superior to the rest and comparable to L. leucocephala. Long-term feeding trials are required with the superior species when used as protein supplements to poor quality diets.

Preparation of Egg White Liquid Hydrolysate (ELH) and Its Radical-Scavenging Activity

  • Noh, Dong Ouk;Suh, Hyung Joo
    • Preventive Nutrition and Food Science
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    • v.20 no.3
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    • pp.183-189
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    • 2015
  • In the present study, an optimum protease was selected to hydrolyze the egg white liquid protein for the antioxidant peptides. Alcalase treatment yielded the highest amount of ${\alpha}$-amino groups (15.27 mg/mL), while the control (no enzymatic hydrolysis) showed the lowest amount of ${\alpha}$-amino groups (1.53 mg/mL). Alcalase also gave the highest degree of hydrolysis (DH) value (43.2%) and was more efficient for egg white liquid hydrolysis than the other enzymes. The Alcalase hydrolysate had the highest radical-scavenging activity (82.5%) at a concentration of 5.0 mg/mL. The conditions for enzymatic hydrolysis of egg white liquid with Alcalase were selected as substrate : water ratio of 2:1. Five percent Alacalse treatment did not show significant (P>0.05) increases of DH and ${\alpha}$-amino nitrogen content after 24 hhydrolysis. Thirty two hour-hydrolysis with 5% Alcalase is sufficient to make antioxidative egg white liquid hydrolysate from egg white liquid. DPPH and ABTS radical-scavenging activities were significantly (P<0.05) higher after enzymatic digestion. These results suggest that active peptides released from egg-white protein are effective radical-scavengers. Thus, this approach may be useful for the preparation of potent antioxidant products.

Identification of Receptor-like Protein for Fructose-1,6-bisphosphatase on Yeast Vacuolar Membrane

  • Ko, Je-Sang
    • BMB Reports
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    • v.33 no.6
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    • pp.448-453
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    • 2000
  • In yeast the key gluconeogenic enzyme, fructose-1,6-bisphosphatase (FBPase), is selectively targeted from the cytosol to the lysosome (vacuole) for degradation when glucose starved cells are replenished with glucose. The pathway for glucose induced FBPase degradation is unknown. To identify the receptor-mediated degradation pathway of FBPase, we investigated the presence of the FBPase receptor on the vacuolar membrane by cell fractionation experiments and binding assay using vid mutant (vacuolar import and degradation), which is defective in the glucose-induced degradation of FBPase. FBPase sedimented in the pellets from vid24-1 mutant after centrifugation at $15,000{\times}g$ for 15 min, suggesting that FBPase is associated with subcellular structures. Cell fractionation experiments revealed that FBPase is preferentially associated with the vacuole, but not with other organelles in vid24-1. FBPase enriched fractions that cofractionated with the vacuole were sensitive to proteinase K digestion, indicating that FBPase is peripherally associated with the vacuole. We developed an assay for the binding of FBPase to the vacuole. The assay revealed that FBPase bound to the vacuole with a Kd of $2.3{\times}10^6M$. The binding was saturable and specific. These results suggest that a receptor for FBPase degradation exists on the vacuolar membrane. It implies the existence of the receptor-mediated degradation pathway of FBPase by the lysosome.

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THE EFFECT OF GINSENOSIDE-TRIOL ON THE POSTOPERATIVE RECOVERY IN GYNECOLOGICAL PATIENTS

  • Chang Yoon Seok;Lee Jin Yong;Kim Chong Woo
    • Proceedings of the Ginseng society Conference
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    • 1978.09a
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    • pp.79-84
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    • 1978
  • Ginseng has been widely used in the Oriental world for more than 2,000 years. Its chemical and pharmacological studies have been published by many investigators of many countries. But its clinical studies have not been performed in satsifactory amount. This study was carried out to evaluate the effect of ginsenoside-triol on the postoperative recovery in 120 cases of gynecological laparotomies. Daily dose of 0.23 gram of ginsenoside-triol was administered orally for three weeks after surgery to 60 cases, and placebo to 60 cases as control. Hemoglobin, hematocrit, leukocyte count, serum total protein, albumin, cholesterol and glucose were studied at pre- and postadministration. At the same time, body weight, blood pressure and subjective symptoms such as appetite, bowel movement and digestion were checked. The results obtained were summarized as follows: 1) The side effect was nil. 2) Hemoglobin and hematocrit Were more increased in treated group than in control group, but the changes were not significant. 3) Serum total protein was more significantly increased in treated group than in control group. 4) Serum cholesterol was significantly less increased in treated group than in control group. 5) Serum glucose level was significantly decreased in both groups, more significantly in control group. 6) Body weight was significantly increased in treated group.

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Proteolysis Analysis and Sensory Evaluation of Fermented Sausages using Strains Isolated from Korean Fermented Foods

  • Chang-Hwan Jeong;Sol-Hee Lee;Hack-Youn Kim
    • Food Science of Animal Resources
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    • v.43 no.5
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    • pp.877-888
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    • 2023
  • We studied the proteolysis and conducted a sensory evaluation of fermented sausages using strains derived from Kimchi [Pediococcus pentosaceus-SMFM2021-GK1 (GK1); P. pentosaceus-SMFM2021-NK3 (NK3)], Doenjang [Debaryomyces hansenii-SMFM2021-D1 (D1)], and spontaneous fermented sausage [Penicillium nalgiovense-SMFM2021-S6 (S6)]. Fermented sausages were classified as commercial starter culture (CST), mixed with GK1, D1, and S6 (GKDS), and mixed with NK3, D1, and S6 (NKDS). The protein content and pH of GKDS and NKDS were significantly higher than those of CST on days 3 and 31, respectively (p<0.05). Sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed that the NKDS had higher molecular weight proteins than the GKDS and CST. The myofibrillar protein solubility of the GKDS and NKDS was significantly higher than that of the CST on day 31 (p<0.05). The GKDS displayed significantly higher pepsin and trypsin digestion than the NKDS on day 31 (p<0.05). The hardness, chewiness, gumminess, and cohesiveness of the GKDS were not significantly different from those of the CST. The GKDS exhibited the highest values for flavor, tenderness, texture, and overall acceptability. According to this study, sausages fermented using lactic acid bacteria (GK1), yeast (D1), and mold (S6) derived from Korean fermented foods displayed high proteolysis and excellent sensory evaluation results.

GROWTH PERFORMANCE AND AMINO ACID DIGESTIBILITIES AFFECTED BY VARIOUS PLANT PROTEIN SOURCES IN GROWING-FINISHING PIGS

  • Moon, H.K.;Kim, J.W.;Heo, K.N.;Kim, Y.H.;Kim, S.W.;Kwon, C.H.;Shin, I.S.;Han, In K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.7 no.4
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    • pp.537-546
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    • 1994
  • This experiment was carried out to compare the effects of six different plant protein sources such as soybean meal, extruded full-fat soybean, canola meal, rapeseed meal, cottonseed meal and perilla meal as a sole protein source of diets on growth performance and amino acid bioavailabilities in growing-finishing pigs. A total of 54 pigs with average 25 kg of body weight were used as experimental subjects for a 65-d feeding trial. Digestion trial was carried out with seven ileal-cannulated pigs. The most rapid rate of weight gain was observed in pigs fed soybean meal and full-fat soybean, the moderate one in pigs fed canola meal and cottonseed meal and the least one in pigs fed rapeseed meal and perilla meal (p<0.005). Feed efficiency was better for groups fed soybean meal and full-fat soybean than other protein meals (p<0.05). The apparent ileal digestibilities of essential amino acids of soybean meal and full-fat soybean (82.5% and 81.6%) were significantly (p<0.05) higher than those of other protein sources (61.2 to 69.4%). Regardless of protein sources, the apparent ileal digestibility of arginine was highest, whereas that of histidine was lowest among essential amino acids. Proline had the lowest digestibility among non-essential amino acids. True amino acid digestibilities tended to be higher than apparent amino acid digestibilities. The differences between true and apparent ileal digestibilities were greater in canola meal, rapeseed meal or cottonseed meal than other protein sources. The differences was greatest in praline except for cottonseed meal. The fecal digestibility appeared to be higher than the ileal digestibility. The differences between fecal and ileal digestibilities were greater in canola meal, rapeseed meal, cottonseed meal and perilla meal than in soybean meal and full-fat soybean. In general, praline was the most disappeared amino acid in the hind gut, while the net synthesis of lysine in the large intestine was observed in all protein sources except perilla meal. It is appropriate that swine feeds should be formulated based on true ileal amino acid digestibility of protein sources for pig's normal growth.