• Parasites, Hosts and Diseases
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• v.27 no.3
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• pp.161-170
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• 1989

The proteases of Toxoplasma gcndii were purified partially and characterisrd for some biochemical properties including various chromatographic patterns, major catalytic classes, and conditions to promote the activity of these enzymes. When Toxoplasma extract was incubated with 3H-casein at various pH, peak hydrolysis of casein was observed at pH 6.0 and at pH 8.5. Proteasfs working at pH 6.0 and at pH 8.5 were purified partially by conventional methods of chromatographies of DE52 anion rxchange, Sephadex G-200 gel permeation, and hydroxylapatite chromatography. Partially purified enzymes were tested by site-specific inhibitors and promotorf. The protease working at pH 6.0 was inactivated by iodoacetamide with LDso of 10-5 M and promoted by dithiothreitol, while the protease working at pH 8.5 was inhibited by phenylmethylsulfonyl fluoride with LD50 of 10-5 M and was Promoted by ATP (excess ATP beyond 2 mM inhibited the activity reversely). The protease of pH 8.5 had the activity of ATPase which might exert the energy to its action. Therefore the former was referred to as a cysteinyl acid protease and the latter, ATP-dependent neutral serine protease.

• Journal of Dairy Science and Biotechnology
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• v.26 no.1
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• pp.27-36
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• 2008

This review was written to introduce updated data on the structure and function of the major milk proteins identified as allergens, the characterization of their epitopes in each allergenic milk proteins, and the reduction of milk protein allergenicity. Most mammalian milk protein, even protein present at low concentration, are potential allergens. Epitopes identified in milk proteins are both conformational(structured epitope) and sequential epitopes(linear epitope), throughout the protein molecules. Epitopes on casein and whey proteins are reported to be sequential epitope and conformational epitopes, respectively. Conformational epitopes on whey protein are changed into sequential epitope by heat denaturation during heat treatment. Several methods have been proposed to reduce allergenicity of milk proteins. Most ideal and acceptable method to make hypoallergenic milk or formula, so far, is the hydrolysis of allergenic milk proteins by enzymes that has substrate specificity, such as pepsin, trypsin, or chymotrypsin. Commercial formulas based on milk protein hydrolysate are available for therapeutic purpose, hypoantigenic formula for infants from families with a history of milk allergy and hypoallergenic formula for infants with existing allergic symptoms.

• Korean Journal of Microbiology
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• v.24 no.2
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• pp.119-126
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• 1986

A periplasmic endoprotease, named protease Pi, was purified to homogeneity from Escherkchia coli by conventional procedure with insulin as substrate. This enzyme degrades insulin and glucagon to trichloroacetic acid-soluble meterials, but shows little or no hydrolysis of bovine serum albumin, casein or globin. Its molecular weight was 110, 000 when determined by gel filtration on Sephacryl S-300 and was 105, 000 when estimated by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate. Thus, it appears to be single polypeptide. This snzyme is metalloprotease, since it is completely inhibited by o-phenanthroline and can be activated by addition of divalent metal cations, such as $Mg^{2+}\;and\;Co^{2+}$. It is destinct from protease Ci, a cytoplasmic insulin degrading enzyme, since protease Pi is localized to the periplasm. Since protease Pi selectively degrades GTP cyclohydrolase I, it appears to play a role in the regulation of pteridine biosynthesis.

• Applied Biological Chemistry
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• v.13 no.3
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• pp.223-229
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• 1970

1. The preparation and some enzymatic properties of crude alkaline protease from a thermophilic mold, Myriococcum sp. was investigated. 2. Optimum pH for the hydrolysis of casein was 9.0 at $50^{\circ}C$ for 10 minutes. Optimum temperature was $55^{\circ}C$ at pH 9.0 for 10 minutes. The enzyme was highly stable at the range of pH 6.0 to 11.0 at $30^{\circ}C$ 3. The alkaline protease in the culture filterate was isolated two fractions by elution column chromatography on DEAE-Cellulose.

• Microbiology and Biotechnology Letters
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• v.24 no.2
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• pp.191-196
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• 1996

The bacteriolytic enzyme produced from Bacillus subtilis SH-1 was purified and characterized, and its molecular weight was determined. The bacteriolytic enzyme activity was increased about 66.5 times via purification with recovery yield of 18.5%. The optimum pH and temperature of this enzyme were 9.0 and 50$\circ$C. The enzyme was stable within a pH range of 6.0-10.0 and unstable above 60 . The molecular weight of the enzyme was estimated to be 23,000 dalton in a form of monomer with no other subunits. Effect of the enzyme on the lysis of bacteria engaged in food posion was tested. The lysis degree was below 31% against Gram negative bacteria and above 48% in Gram positive bacteria. The values higher than 73% were obtained against Vibrio sp. and Listeria sp. As the turbidity of dissolved peptidoglycan clecreases, the free amino group levels were increased. And, based on hydrolysis of casein, this enzyme was thought to be an endopeptidase.

• Journal of Nutrition and Health
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• v.26 no.5
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• pp.585-592
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• 1993

In order to investigate the hypocholesteremic effect of soybean perptides, soybean protein(ISP), casein(CNP), their peptic hydrolyzates fractionated by acid precipitation at different pH's(SHT, SH8, SH6, SH4, CHT, CH5, CH4) and amino acid mixtures of the same composition as the proteins(SAA, CAA) were fed to rats and the concentration of serum cholesterol was measured. Then in vitro digestibility and molecular weight distribution of the peptides by pepticpancreatic hydrolysis was measured by FPLC. The lower the in vitro digestibility of peptides is, the lower the concentration of serum cholesterol becomes(r=0.986) and the higher the ratio of macropeptides is, the lower the concentration of serum cholesterol becomes(r=-0.932) in rats. These results suggest that the in vitro digestibility of peptides has close relationship to the concentration of serum cholesterol in rats and non-digestible meacropeptides or polypeptides especially more than 1 kDa, formed through digestion in gut, may lower the serum cholesterol in rats.

• Asian-Australasian Journal of Animal Sciences
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• v.25 no.9
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• pp.1269-1275
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• 2012

An experiment was conducted to study the effect of soluble protein supplements on concentration of soluble non-ammonia nitrogen (SNAN) in the liquid phase of ruminal (RD) and omasal digesta (OD) of Korean native steers, and to investigate diurnal pattern in SNAN concentration in RD and OD. Three ruminally cannulated Korean native steers in a $3{\times}3$ Latin square design consumed a basal diet of rice straw and corn-based concentrate (control), and that supplemented (kg/d DM basis) with intact casein (0.24; IC) or acid hydrolyzed casein (0.46; AHC). Ruminal digesta was sampled using a vacuum pump, whereas OD was collected using an omasal sampling system at 2.0 h intervals after a morning feeding. The SNAN fractions (free amino acid (AA), peptide and soluble protein) in RD and OD were assessed using the ninhydrin assay. Concentrations of free AA and total SNAN in RD were significantly (p<0.05) lower than those in OD. Although free AA concentration was relatively high, mean peptide was quantitatively the most important fraction of total SNAN in both RD and OD, indicating that degradation of peptide to AA rather than hydrolysis of soluble protein to peptide or deamination may be the most limiting step in rumen proteolysis of Korean native steers. Diurnal variation in peptide concentration in OD for the soluble protein supplemented diets during the feeding cycle peaked 2 h post-feeding and decreased thereafter whereas that for the control was relatively constant during the entire feeding cycle. Diurnal variation in peptide concentration was rather similar between RD and OD.

• Microbiology and Biotechnology Letters
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• v.18 no.5
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• pp.496-500
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• 1990

When pepsin was used at a concentration of 8 mglml for hydrolysis of 0.02% casein, inhibitory activity of this inhibitor was proportional to a inhibitor concentration of 20 ${\mu}g$/ml, and fifty percent inhibition ($IC_{50}$) was observed to be 15 ${\mu}g$/ml. The inhibitor was pH-stable at pH range of 5-9 at $100^{\circ}C$ for 10 minutes and thermo-stable at pH 7.0 at $100^{\circ}C$ to give 100% activity for 20 minutes. The formation of pepsin-inhibitor complex was confirmed by sephadex 6-25 gel filtration and type of inhibition was determined as non-competitive inhibition by Lineweaver-Burk plot. The inhibitor strongly inhibited acid proteases such as pepsin and renin, and it was soluble in methanol very well. On TLC analysis of silicagel 60 using various sohent systems, the inhibitor gave a single spot at Rf range 0.4-0.6. From the result of IR spectrum and color reaction (Rydon-Smith, Biuret), this inhibitor was considered as peptide substance. Melting point and elemental contents were 220-$230^{\circ}C$, and C 50.61%-H 8.02%-N 9.34% (found), respectively.

• Food Science of Animal Resources
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• v.35 no.3
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• pp.360-369
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• 2015

Milk proteins have many potential sequences within their primary structure, each with a specific biological activity. In this study, we compared and investigated the bioactivities of hydrolysates of the domestic (A, B) and imported (C, D) skim milk powders generated using papain digestion. MALDI-TOF analysis revealed that all milk powder proteins were intact, indicating no autolysis. Electrophoretic analysis of hydrolysates showed papain treatment caused degradation of milk proteins into peptides of various size. The antioxidant activity of the hydrolysates, determined using 2,2-azino-bis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) and total phenolic contents (TPC) assays, increased with incubation times. In all skim milk powders, the antioxidant activities of hydrolysates were highest following 24 h papain treatment (TPC: A, 196.48 μM GE/L; B, 194.52 μM GE/L; C, 194.76 μM GE/L; D, 163.75 μM GE/L; ABTS: A, 75%; B, 72%; C, 72%; D, 57%). The number of peptide derived from skim milk powders, as determined by LC-MS/MS, was 308 for A, 283 for B, 208 for C, and 135 for D. Hydrolysate A had the highest antioxidant activity and the most potential antioxidant peptides amongst the four skim milk powder hydrolysates. A total of 4 β-lactoglobulin, 4 α_s1-casein, and 56 β-casein peptide fragments were identified as potential antioxidant peptides in hydrolysate A by LC-MS/MS. These results suggest that domestic skim milk could have applications in various industries, i.e., in the development of functional foods.

• Korean Journal of Food Science and Technology
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• v.21 no.4
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• pp.492-497
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• 1989

The antioxidant effects against linoleic acid of various protein hydrolysates from fish protein, defatted soybean cake, egg albumin and casein were investigated. Each protein hydrolysate by enzyme hydrolysis exhibited the antioxidative effects by addition of 5mg and 10mg per 1g linoleic arid. Especially, egg albumin and fish protein hydrolysates had a great antioxidative effects. The protein hydrolysates indicated the synergitic effects with ${\alpha}-tocopherol$, and indicated scavenging effects toward metal ion $(Fe^{3+},\;Cu^{2+})$ as prooxidants.