• The Korean Journal of Food And Nutrition
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• v.8 no.4
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• pp.335-343
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• 1995

In order to enhance the utility of alaska-pollack, the optimum conditions for the preparation of pronase hydrolysate. The optimum temperature and pH for the hydrolysis of alaska-pollack by pronase were 4$0^{\circ}C$ and pH 7.0. The reaction time and enzyme concentration were 4 hr and 1,000 units per g of substrate. Under the above optimum conditions alaska-pollack was hydrolysed by pronase yielding a hydrolytic degree of about 89eye. The bitterness and hyrophobicity of pronase hydrolysate were decreased with increasing reaction time. Hydrophobic amino acids(Tyr, Met, Ala, flu, Leu, and Phe) were increased for 2 hr, but fur thor hydrolysis was showed decrease of hydrophobic amino acids content. Palatable amino acids (Asp, Glu, Pro, Ser, Thr and Gly) were increased with hydrolysis time.

• Journal of the Korean Society of Food Science and Nutrition
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• v.21 no.5
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• pp.513-518
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• 1992

Optimum conditions for enzymatic hydrolysis of purified rapeseed(Brassica napus var. Youngsan) protein were investigated. Pronase showed higher activity in the hydrolysis of rapeseed protein than that of alcalase and neutrase. Preheated treatment of the rapeseed protein decreased the activity of pronase. The degree of hydrolysis of rapeseed protein was greater in distilled water than in phosphate buffer solution. Degree of hydrolysis was reached in steady state after 1 hr. Optimum conditions of the hydrolysis of the rapeseed protein were $40^{\circ}C$ in reaction temperaturem pH 8.0 in substrate solution, 1/100 (w/w) in the ratio of enzyme to substrate and 1% (w/v) in substrate concentration for pronase, respectively. At the optimum hydrolysis conditions, Km value was 3.48% (w/v).

• Applied Biological Chemistry
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• v.35 no.5
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• pp.339-345
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• 1992

In order to enhance the processing quality and utility of alaska-pollack meat, the optimum conditions for the preparation of pronase hydrolysate and the synthesis of plastein were investigated. The optimum temperature and pH for the hydrolysis of alaska-pollack by pronase were $40^{\circ}C$ and pH 7.0. The reaction time and enzyme concentration were 4 hr and 1,000 units per g of substrate. Under the above optimum conditions alaska-pollack was hydrolyzed by pronase yielding a hydrolytic degree of about 89%. Pronase hydrolysate was employed as substrate for plastein synthesis. The 30% pronase hydrolysates were adjusted to pH 7 for fruit-bromelain and pH 5 for stem-bromelain, and then plastein were synthesized by 1% bromelain at $40^{\circ}C$ for 24 hr. The plasteins synthesized by fruit- and stem-bromelain were consisted of peptides having average peptide length of 22.6 and 20.8 under the optimum synthetic conditions. The plastein synthesis reaction reduced considerably the bitterness of pronase hydrolysate.

• Korean Journal of Food Science and Technology
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• v.8 no.4
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• pp.253-259
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• 1976

Two kinds of sepharose-bound pronases were successfully prepared. The immobilized pronase, directly coupled to cyanogen-bromide activated sepharose, retains 22.6% of original specific activity against casein. However, ${\omega}-aminoalkyl$ sepharose immobilized pronases, in which extension arms of ${\omega}-aminoalkyl$ group $(number\;of\;-CH_2-\;is\;8,\;10,\;and\;12)$ are used, retain almost 100% of original specific activity. Studies of enzyme stability, pH dependence, temperature dependence, and Km values are presented.

• Clinical and Experimental Reproductive Medicine
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• v.27 no.4
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• pp.345-351
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• 2000

Objective: Hatching of the blastocyst from the zona pellucida (ZP) is a key event in mammalian implantation. In vivo, two factors have been identified as possible mediators of hatching: lysis of the ZP by substances elaborated either from the embryo or female reproductive tract and pressure exerted on the zona by expansion of the blastocyst. Two methods of zona manipulation were already in use to enhance the ability of embryos to hatch: mechanical PZD and chemical ZD by acidic Tyrode's solution. But several controversies of each method have been reported. The purpose of this study was to investigate the effect of pronase for mouse embryo hatching. Methods: Mouse embryos were obtained following ovulation induction of $F_1$ animals. Fresh and cryo-thawed morula embryos were exposed to 0.5, 1.0, 2.0, 5.0 ${\mu}g/ml$ pronase in Ham's F10 for 72 hrs. Main outcome measures were the rates of partial hatching and completely hatched blastocysts, and cell number of it. Results: In fresh and cryo-thawed group, the rates of completely hatched blastocyst were significantly higher in 5 ${\mu}g/ml$ pronase treatment group than control group. There was no difference in completely hatched blastocyst total cell number between pronase treatment group and control group. This suggest that pronase treatment did not harmful in mouse embryo development. In pronase treatment group, zona pellucida were thinner than control group. Conclusion: The addition of pronase to culture media may accelerate the hatching of embryo. So, enzymatic treatment of the zona may provide a valuable and effective assisted hatching technique for human in-vitro fertilization-embryo transfer.

• KSBB Journal
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• v.6 no.4
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• pp.327-336
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• 1991

A study on the optimum hydrolysis conditions of fish skin through the aid of enzymes and the development of a natural seasoning using the hydrolysate has been carried out for the effective utilization of fish skin. Using the "pH-drop" techniques the collagenase and pronase were identified as most suitable for this purpose. The $K_m$ and $V_{max}$ values of pronase were 1.82 mgN/ml and 0.06 mgN/mL/min, respectively. The hydrolysis conditions of the cod skin for the pronase were as follows: reaction temperature, $50^{\circ}C$; reaction time, 3hrs; pH 6; enzyme concentration, 0.03%. The degree of hydrolysis at these conditions was 76.8%. But after hydrolyzing cod skin with collagenase for 1hr, when the pronase was treated, the degree of hydrolysis was 83.13%. The molecular weight of the hydrolysate was 8,000 daltons. Among the amino acids in the hydrolysate, glycine(27.95%), glutamic acid(10.94%), proline(7.39%), aspartic acid(9.47%) and serine(7.39%) were responsible for 64.23% of the total amino acids. But valine, methionine, isoleucine, leucine, phenylalanine and histidine having a bitter taste were only 13.05%. From the results of the sensory evaluation, the imitation sauce which was made of 20% fermented soy sauce prepared from the hydrolysate was at least similar to the traditional soybean sauce in product quality. The complex seasoning containing 31.7% of the hydrolysate was nearly equal to complex seasonings on the market, too.

• Clinical and Experimental Reproductive Medicine
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• v.28 no.3
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• pp.235-245
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• 2001

Objectives: The purpose of this present study was to compare mouse embryo development in 3 commercial media and hatching competence of mouse embryo with or without enzymatic treatment. Methods: Collected 375 mouse embryos were divided into three groups, and then cultured in IVF-20 (G2), Medicult IVF (M3), P-1 (blastocyst M), respectively. Three day mouse morulae were cultured in G2 media treated with pronase. The results were analyzed using Chi-square test, and considered statistically significant when p<0.01. Results: The developmental rate of 2 cell mouse embryo after 72 hours was highest in IVF-20 (G2) among conventional 3 media. The hatching rate of mouse morulae was low when clultured in G2 media without pronase during 48 hours. However, it was higher when cultured in media treated with $1{\mu}g/ml$, $2.5{\mu}g/ml$, $5{\mu}g/ml$ pronase, respectively. Conclusions: Using good media and digestion of zona pellucida with enzymatic treatment improve development and hatching rate of embryo. Therefore, implantation and pregnancy rate could be improved.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.49 no.5
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• pp.594-599
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• 2016

Infection with HIV (Human immunodeficiency virus), over time, develops into acquired immunodeficiency syndrome (AIDS). The development of non-toxic and effective anti-HIV drugs is one of the most promising strategies for the treatment of AIDS. In this study, we investigated the anti-HIV-1 activity of gelatin hydrolysates from Alaska pollack skin. Gelatin hydrolysates were prepared using four enzymes (alcalase, flavourzyme, neutrase, and pronase E). Among these, the pronase E gelatin hydrolysate was found to inhibit HIV-1 infection in the human T cell-line MT4. It exhibited inhibitory activity on HIV-1_IIIB-induced cell lysis, reverse transcriptase activity, and viral p24 production at noncytotoxic concentrations. Moreover, it decreased the activation of matrix metalloproteinase-2 (MMP-2) in vitro. Because HIV infection-induced activation of MMP-2 can accelerate collagen resolution and collapse of the immune system, pronase E gelatin hydrolysate might prevent the activation of MMP-2 in cells, resulting in collagen stabilization and immune cell homeostasis consistent with anti-HIV activation. These results suggest that pronase E gelatin hydrolysate could potentially be incorporated into a novel therapeutic agent for HIV/AIDS patients.

• Bulletin of the Korean Chemical Society
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• v.20 no.11
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• pp.1299-1302
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• 1999

Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) was used to demonstrate cross-linking of peptides induced by transglutaminase. The presence of ε-( Υ-glutamyl)lysine isopeptide cross-link in the acid hydrolysate of the cross-linking reaction mixture was also demonstrated by MALDI-TOF-MS without prior separation. MALDI-TOF-MS quickly provided peptide mass maps after pronase digestion of the cross-linked peptide adduct, which enabled us to monitor the hydrolytic sequence. Pronase appears to preferentially hydrolyze peptide bonds distant from the cross-link before hydrolyzing peptide bonds around the cross-link. The results suggest that pronase digestion followed by MALDI-TOF-MS could be used for determination of amino acid sequence around a modification site.

• Korean Journal of Veterinary Research
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• v.29 no.4
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• pp.445-455
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• 1989

The proteins of the ruminant erythrocyte membranes were analysed by polyacrylamide gel electrophoresis in sodium dodecyl sulfate, and their relations to the slow erythrocyte sedimentation rate(ESR) of the ruminants were investigated by treating the erythrocytes with proteinases such as trypsin, chymotrypsin and pronase, and glycosidases such as neuraminidase and galactosidase. Protein content in the erythrocyte membrane was $2.85{\pm}0.28$ in human, $3.60{\pm}0.41$ in Korean cattle, $3.71{\pm}0.36$ in Holstein, $4.13{\pm}0.83$ in Korean native goat and $3.94{\pm}0.56mg/ml$ in sheep, showing higher in ruminant animals than in human(p<0.01). Although the general protein profiles of the ruminant erythrocyte membranes were almost similar to that of human, all the ruminant erythrocyte membranes showed one additional protein band, called band-Q in the previous report on proteins of bovine erythrocyte membrane, which migrated electrophoretically to the mid position between band-2 and band-3 in human erythrocyte membranes. The glycoprotein profiles of ruminant erythrocyte membranes revealed by periodic acid Schiff(PAS) stain showed a marked difference from that of human. The PAS-1(glycophorin) and PAS-2(sialoglycogrotein) present in human erythrocyte membranes were almost absent from the ruminant animals. Instead, a strong PAS-positive band near the origin of the electrophorograms, which was named as PAS-B in the previous report on proteins of bovine erythrocyte membranes, was shown in the ruminant animals except sheep. In addition, the erythrocyte membranes of Korean native goat and sheep showed a moderate PAS-negative band near the tracking dye of the electrophorograms, which was named as PAS-G in this study. In the erythrocyte treated with the enzymes, the migration of each protein fracture of erythrocyte membranes in response to each enzyme was diverse according to different species or breed of ruminant animals. Among others, band-Q present in ruminants was slightly or moderately decreased by trypsin-, chymotrypsin-, and pronase- treatments of the erythrocytes, but not only in sheep. It was particularly noticeable that PAS-B, a fraction of glycoprotein, present in ruminants except sheep, was better digested by proteinases than by glycosidases, showing remarkable increase(p<0.01) of the ESR in accord with complete digestion(disappearance) of the PAS-B band by pronase, trypsin or chymotrypsin treatment of erythrocytes. In sheep, there was almost no any response to the various enzymes in general protein and glycoprotein profiles of the erythrocyte membranes except PAS-G, which was markedly decreased by pronase treatment of the erythrocytes. Nevertheless, the ESRs were accelerated in erythrocytes treated with pronase, trypsin, chymotrypsin and neuraminidase. Erythrocyte osmotic fragility was increased in erythrocytes treated with only pronase among five enzymes in all the human and ruminant animals used in this study.