• Title/Summary/Keyword: Trypsin inhibitor

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PIGEON PEA AS A RUMINANT FEED

  • Cheva-Isarakul, B.
    • Asian-Australasian Journal of Animal Sciences
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    • v.5 no.3
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    • pp.549-558
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    • 1992
  • The study on the potential use of pigeon pea (PP) as a ruminant feed was carried out with sheep in 3 experiments. Digestibility of dry pigeon pea leaves (PPL) and pigeon pea seeds (PPS) determined by differential and regression methods respectively, with rice straw (RS) as a basal diet, revealed that PPS contained higher nutritive value and palatability than PPL. On dry matter (DM) basis, PPL and PPS contained 19.8 and 20.0% CP, 7.3 and 2.3% EE, 6.0 and 4.4% ash, 61.1 and 51.7% NDF, and 29.4 and 17.5% ADF, respectively. The trypsin inhibitor activity in the seed was 3 times of that in the leaves (19.5 vs 7.0 mg TIA/g DM). The digestibility of PPL and PPS were 50.2 and 72.2% in DM, 52.7 and 73.3% in OM, 51.0 and 65.1% in CP respectively. DM intake as well as the digestibility of most nutrients increased with the increasing level of PPS. Digestible energy (DE), Total digestible nutrient (TDN) and N-balance of sheep fed solely PPS, estimated by regression method, was 3.2 kcal/g, 71.1% and 6.3 g/d respectively. Pigeon pea seeds can be well used to substitute soybean meal in concentrate rations for ruminants or directly supplemented to low quality roughages.

Studies on Rabbit Serum Inhibitor of Avian Infectious Bronchitis Virus (전염성기관지염(傳染性氣管支炎) 바이러스에 대한 가토혈청억제물질(家兎血淸抑制物質)에 관한 연구(硏究))

  • Yoo, Tai Suck
    • Korean Journal of Veterinary Research
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    • v.5 no.1
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    • pp.43-57
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    • 1965
  • It has been-reported that rabbit serum exhibit an inhibitory action on avian infectious bronchitis virus in embryonating chicken embryo. In this thesis, the biological, serological, physical and chemical properties of normal rabbit serum on the effect of the virus propagation were studied. Throughout the studies, the following experimental results 'were obtained and summarized here. 1. An inhibitory action of rabbit serum on avian infectious bronchitis vrius is due to the normal serum constituents. 2. The nature of the neutralization between normal rabbit serum and the virus is similar to that of the specific antiserum and the virus. 3. Rabbit serum, heat inactivated at $56^{\circ}C$, for 30 minutes, showed its average $log_{10}El,D_{50}Nl$ of 3.7. 4. The inhibitory compound present in the normal rabbit serum is inactivated by means of 5 per cent trypsin, 0.01 M potassium periodate, and absorbed to zymosan. 5. The inhibitory compound was not affected by 0.05 M trichloroacetic acid and 0.005M $KH_2PO_4$. 6. The higher the temperature of heat inactivation of rabbit serum caused the lesser the neutralizing effect on the virus. Heating the serum at $66^{\circ}C$, for 30 minutes brought about a complete loss of the neutralizing index of the serum. 7. No ions, as a cofactor, was incorporated to the inhibitory action of rabbit serum on the virus. 8. The inhibitory compound amays be found in a fraction of serum globulin.

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Tissue- and Reproductive Organ-specific Expression of Protease Nexin-1 in Sprague-Dawley Rat (흰쥐에서 단백질 분해효소 저해제, Nexin-1의 조직 및 생식기관 특이적 유전자 발현)

  • 고정재;김남근;김진규;최명진;정형민;서승염;김윤희;이현환;차광열
    • Development and Reproduction
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    • v.2 no.2
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    • pp.135-140
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    • 1998
  • Protease Nexin-1 (PN-1) inhibits the activity of several serine proteases including thrombin, urokinase (uPA)-type plasminogen activator and trypsin. Tissue- and reproductive organ-specific mRNA levels of the PN-1 were investigated in Sprague-Dawley adult rat. PN-1 mRNA expression in rats was found in brain (forebrain, hindbrain), heart, liver, lung, ovary and oviduct. The level of PN-1 mRNA in male and female among the tissues was the highest in forebrain of the female. PN-1 expression in reproductive organs was found only in ovary and oviduct. These results suggest that PN-1 expression is dependent on the sex and may be related to folliculogenesis and early embryogenesis.

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High Temperature-Cooking Effects on Protein Quality of Fish Extracts

  • Ryu, Hong-Soo;Moon, Jeong-Hae;Hwang, Eun-Young;Yoon, Ho-Dong
    • Preventive Nutrition and Food Science
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    • v.3 no.3
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    • pp.241-247
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    • 1998
  • Fish extracts were processed at high temperature (136.7 ~14$0^{\circ}C$) for possible use as functional food ingredients. Raw fish meats and those hydrothermal extracts were compared with respect to in vitro and in vivo protein qualities. 95% of fat inraw meats was reduced in extracts but there were not remarkable changes in other macronutrients in freeze-dried extracts. Most of essential amino acids were decreased significantly but two times more proline and glycine were detected in extracts. High temperature cooking resulted 2.1 ~3.7 times of higher total free amino acid content infish extracts compared iwth raw meat, and taurine and glutamic acid were increased especially. Severe protein damages were occurred when invitro protein quality indices such as availblae lysine, hydrophilic browing, trypsin inhibitor formation and in vitro protein digestibility were measured on fish extracts. In vivo protein qualities were also strongly influenced by high temperature ; however rat-body-weight gain was nearly zero during PER assay, and rat PER or NPR of fish extracts were significantly lower (p<0.001) than those of cotnrol (ANRC casein) and original raw fish meats.

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Tryptic Digestion and Cytochalasin B Binding Assay of the Human HepG2-Type Glucose Transporter Expressed in Spodoptera frugiperda Clone 21-AE Cells

  • Lee Chong-Kee
    • Biomedical Science Letters
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    • v.11 no.1
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    • pp.57-61
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    • 2005
  • The number of sites at which a protein can be readily cleaved by a proteolytic enzyme is greatly influenced by its three-dimensional structure. For native, properly-folded proteins both the rate of cleavage and number of sites at which cleavage takes place are usually much less than for the denatured protein. In order to compare the tertiary structure of recombinant HepG2 type glucose transporter with that of its native counterpart in the erythrocyte, the pattern of tryptic cleavage of the protein expressed in insect cell membranes was therefore examined. After 30 minutes digestion, a fragment of approximate Mr 19,000-21,000 was generated. In addition to this, there were two less intensely stained fragments of apparent Mr 28,000 and 17,000. The pattern of labelling was similar up to 2 hours of digestion. However, the fragments of Mr 19,000-21,000 and Mr 17,000 were no longer detectable after 4 hours digestion. The observation of a very similar pattern of fragments yielded by tryptic digestion of the HepG2 type transporter expressed in insect cells suggests that the recombinant protein exhibits a tertiary structure similar if not identical to that of its human counterpart. Also, the endogenous sugar transporter(s) present in Sf21 cells did not bind cytochalasin B, the potent transporter inhibitor. Therefore, the baculovirus/Spodoptera frugiperda (Sf) cell expression system could be very useful for production of large amounts of human glucose transporters, heterologously.

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Purification and characterization of β-secretase inhibitory peptide from sea hare (Aplysia kurodai) by enzymatic hydrolysis

  • Lee, Jung Kwon;Kim, Sung Rae;Byun, Hee-Guk
    • Fisheries and Aquatic Sciences
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    • v.21 no.5
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    • pp.13.1-13.8
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    • 2018
  • Amyloid plaque, also called senile plaque, the product of aggregation of ${\beta}$-amyloid peptides ($A{\beta}$), is observed in brains of the patients with Alzheimer's disease (AD) and is one of the key factors in etiology of the disease. In this study, hydrolysates obtained from the sea hare (Aplysia kurodai) were investigated for ${\beta}$-secretase inhibitory peptide. The sea hare's muscle protein was hydrolyzed using six enzymes in a batch reactor. Trypsin hydrolysate had highest ${\beta}$-secretase inhibitory activity compared to the other hydrolysates. ${\beta}$-secretase inhibitory peptide was separated using Sephadex G-25 column chromatography and high-performance liquid chromatography on a C18 column. ${\beta}$-secretase inhibitory peptide was identified as eight amino acid residues of Val-Ala-Ala-Leu-Met-Leu-Phe-Asn by N-terminal amino acid sequence analysis. $IC_{50}$ value of purified ${\beta}$-secretase inhibitory peptide was $74.25{\mu}M$, and Lineweaver-Burk plots suggested that the peptide purified from sea hare muscle protein acts as a competitive inhibitor against ${\beta}$-secretase. Results of this study suggest that peptides derived from sea hare muscle may be beneficial as anti-dementia compounds in functional foods or as pharmaceuticals.

The Physicochemical Properties of $\alpha$-Amylase Inhibitors from Black Bean and Naked Barey in Korea (한국산 검정콩 및 쌀보리 $\alpha$-Amylase 저해물질의 이화학적 특성)

  • 심기환;문주석;배영일
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.27 no.3
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    • pp.367-375
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    • 1998
  • The physicochemical properties of the $\alpha$-amylase inhibitors from black bean and naked barley is Korea were investigated. Preincubation time for maximum inhibition was 30min and no activity change was seen after that time. Optimum pH of the $\alpha$-amylase inhibitors from the black bean and naked barley was pH 7.0 and the inhibitory activities were stable in the range of pH 6.0~8.0 in both phosphate and Tris-HCI buffer solutions. Both inhibitors maintained more than 50% of activity after incubation for 17 min at 7$0^{\circ}C$. The inhibitors from the black bean and naked barley maintained more than 50% of activities after treatment for 40 min and 30 min with pepsin, and 30 min and 50 min with trypsin, respectively. Both inhibitors functioned via a noncompetitive mechanism and were active against porcine pancreatic and human salivary $\alpha$-amylases. The activities of both inhibitors were linear for the ionic stength ranging from 0 to 0.9. The addition of 70 mM maltose to the reaction mixture caused a maximum increase in the relative activities of both inhibitors, but it did not affect the dissociation of the EI complex. The activities of both inhibitors were significantly enhanced by adding 1mM of K+ or Mg2+.

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Anti-rheumatoidal Effect of Sulfuretin Isolated from the Heartwood of Rhus veniciflua in Rats and Mice

  • Choi, Jong-Won;Yoon, Byung-Jae;Keun-Huh;Park, Kun-Young;Lee, Kyung-Tae;Park, Hee-Juhn
    • Preventive Nutrition and Food Science
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    • v.7 no.4
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    • pp.347-352
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    • 2002
  • The present study was undertaken to evaluate the anti-rheumatoidal arthritis effect of the R. verniciflua heartwood extract, its EtOAc fraction, and its primary flavonoids, sulfuretin and fustin. All test samples showed variably significant inhibitory effects on hind paw edema and trypsin inhibitor activity induced by Freund's complete adjuvant reagent (FCA reagent), and on vascular permeability caused by acetic acid. Treatment with 10 mg/kg (i.p.) sulfuretin for seven days inhibited edema formation by 54.2$\pm$3.0%. Test samples, especially sulfuretin, shifted the values of biochemical parameters such as serum-cholesterol, serum-triglyceride and serum-total protein toward the normal and restored the numbers of leucocytes and platelets. These results suggest that the heartwood of R. verniciflua reduces immunological injuries caused by FCA reagent provides evidence that suluretin is an active anti-rheumatoid arthritis agent.

Purification and Characterization of a Collagenase from the Mackerel, Scomber japonicus

  • Park, Pyo-Jam;Lee, Sang-Hoon;Byun, Hee-Guk;Kim, Soo-Hyun;Kim, Se-Kwon
    • BMB Reports
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    • v.35 no.6
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    • pp.576-582
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    • 2002
  • Collagenase from the internal organs of a mackerel was purified using acetone precipitation, ion-exchange chromatography on a DEAE-Sephadex A-50, gel filtration chromatography on a Sephadex G-100, ion-exchange chromatography on DEAE-Sephacel, and gel filtration chromatography on a Sephadex G-75 column. The molecular mass of the purified enzyme was estimated to be 14.8 kDa by gel filtration and SDS-PAGE. The purification and yield were 39.5-fold and 0.1% when compared to those in the starting-crude extract. The optimum pH and temperature for the enzyme activity were around pH 7.5 and $55^{\circ}C$, respectively. The $K_m$ and $V_{max}$ of the enzyme for collagen Type I were approximately 1.1 mM and 2,343 U, respectively. The purified enzyme was strongly inhibited by $Hg^{2+}$, $Zn^{2+}$, PMSF, TLCK, and the soybean-trypsin inhibitor.

Effects of Sperm Extracts on Sperm - Egg Binding in Mouse (생쥐의 정자 추출물이 정자-난자의 결합에 미치는 영향)

  • Kim, Moon-Kyoo;Gye, Myung-Chan;Choi, Kyoo-Wan;Yoon, Hyun-Soo;Kim, Jong-Heup
    • Clinical and Experimental Reproductive Medicine
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    • v.18 no.1
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    • pp.23-34
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    • 1991
  • In order to study the sperm-egg interaction during fertilization process in mouse, the effects of sperm concentration, the duration of capacitation and insemination, the stages of maturation and development of eggs, and sperm extracts and BSA on sperm binding to egg were examined. Sperm-egg binding was increased depending on sperm concentration within the range of $10^3-10^6$ sperm/ml. It showed the most numbers of sperm-egg binding at 60min from the beginning of preincubation(capacitation) and insemination, respectively. During sperm capacitation, sperm-egg binding inhibitor was released from sperm into the incubation medium. Sperm extracts containing trypsin-like enzyme which is secreted through the acrosome reaction increased the binding. BSA in the culture medium showed a positive effect on the binding. It is suggested that physicochemical alterations of zona pellucida in the process of maturation and fertilization of eggs leaded to inhibition of sperm-egg binding.

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