• 대한화장품학회지
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• 제26권1호
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• pp.81-92
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• 2000

Development of stabilized enzyme was attempted for cosmetic applications. Papain, a proteolytic enzyme, was stabilized through conjugation with a soluble carbohydrate biopolymer, SC-glucan$^{TM}$ . With a novel structure of the conjugation site, stability of the enzyme was significantly enhanced such that more than 90% of the initial activity retained after a month storage at 45$^{\circ}C$, while no activity were detected in native enzyme or enzyme simply mixed with SC-glucan$^{TM}$ after the storage. Conjugation with SC-glucan$^{TM}$ not only extended the half-life of the enzyme on storage at higher temperature, but was also found to protect enzymes against some components contained in cosmetic products for skin care. Cosmetic lotion containing 1 % papain conjugate was more effective and less irritative in exfoliating stratum corneum of human skin than the lotion containing 5% lactic acid, one of the current popular exfoliating agents.gents.

• 한국축산식품학회지
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• 제20권1호
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• pp.8-14
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• 2000

In order to examine physicochemical gelation behavior of ultra high temperature(UHT) pasteurized milk during storage at 4$^{\circ}C$ and 25$^{\circ}C$, pH, electrophoresis, alcohol test, sialic acid contents and free amino groups contents were biweekly determined. The pH of UHT pasteurized milk decreased with increasing storage time. Gelation of the UHT milk occured faster at 25$^{\circ}C$ than at 4$^{\circ}C$ with larger decreasing rate of pH. The alcohol test showed positive results at lower pH than 6.5, which could indicate the casein instability and beginning of gelation. The electrophoretic patterns showed a decrease in the concentrations of all caseins. Degradation of k-casein was faster in all cases, while $\alpha$-casein and $\beta$-casein were also extensively degraded later. The sialic acid contents of the samples increased gradually during storage, and the increasing rate was higher before gel formation. The free amino groups of the samples increased gradually during storage. The increasing rate of free amino groups was faster at 25$^{\circ}C$ than at 4$^{\circ}C$. The samples stored at 25$^{\circ}C$ gelled earlier than those stored at 25$^{\circ}C$, with corresponding increase of free amino groups. The residual proteolytic enzymes, which survived during the UHT heat treatments and were reactivated during storage, could be responsible for UHT pasteurized milk gelation during storage. It is assumed that proteolytic degradation of caseins followed by aggregation would be attributable to complicated reaction mechanism.

• 한국가축번식학회지
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• 제17권2호
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• pp.93-101
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• 1993

These experiments were carried out to investigate whether zona hardening affect the efficiency of in vitro fertilization in mouse oocytes. The soluble properties for zona pellucida of oocytes matured in vivo, aged oocytes, and ovarian oocytes matured in vitro have been analyzed with proteolytic enzyme, 3mg/ml of $\alpha$-chymotrypsin. The mean solubility(t50) for the zona of unfertilized oocytes, oocytes not fertilized at the first inseminati and in vitro produced zygotes were 10.1, 20.3 and 32.3min., respectively. The t50 for zona lysis of fertilized oocytes was significantly difference than those observed for unfertilized oocytes and oocytes not fertilized at the first insemination(P<0.01). In addition, the t50 of zona in ovulated oocytes with and without cumulus cells incubated for 0, 3, 6, 9 and 12 hr in vitro, t50 were 13.9, 11.1, 20.7 and 28.0min., and 22.3, 21.0, 30.0 and 33.5min., respectively. In these experiments, the zona pellucida showed a gradual increase in resistance to dissolution by $\alpha$-chyjotrypsin with in vitro aging for more than 6 hrs. This effect was greater in cumulus-free as compared to cumulus-intact oocytes. Finally, in cumulus-intact and cumulus-free ovarian oocytes matured for 0, 5, 10 and 15 hr in vitro the t50 of zona pellucida were 3.0, 10.6, 18.4 and 24.5 min., and 3.0, 14.0, 26.2 and 32.0 min., respectively. Clear differences in solubility between the zona pellucida of oocytes matured in vivo and in vitro. This data were found suggest that under in vitro conditions there is a gradual change in the soluble properties of the zona pellucida, particularly in the absence of the cumulus cells.

• 한국미생물·생명공학회지
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• 제30권2호
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• pp.184-188
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• 2002

Lactobacillus sake는 냉장온도(2-4 ℃)이하에서도 성장이 가능한 통성혐기성균으로서, sausage와 진공포장육의 제조에 관여하며, 동양에서는 청주의 발효와 김치의 숙성에도 큰 비중을 차지하는 것으로 알려지고 있다. 그러나 L. sake는 이들 식품의 제조시 과도한 발효의 진행으로 식품을 변질시켜 가치를 저하시키기도 한다. 본 연구에서는 L. sake의 생육을 효과적으로 저지하는 박테리오신을 생산하는 미생물을 김치로부터 분리하여 Lactobacillus sp. Oh-B3로 명명하였다. 최대의 박테리오신을 생산하기 위한 분리균의 배양조건은 초기 pH를 8.0으로 조절한 MRS배지에 접종하여 25℃에서 24시간동안 배양했을 때 최대의 박테리오신을 생산하였다. 분리균의 배양액을 원심분리하여 균체를 제거한 상등액에 ammonium sulfate를 첨가한 뒤, 얻어진 침전물을 투석하고 여과한 뒤, -20℃에 보관하며 조박테리오신 용액으로 사용하였다. 분리균이 생산하는 박테리오신은 pH 2.0-9.0의 넓은 범위에서 비교적 안정한 활성을 나타내었으며, 그람음성균에 대해서는 항균활성을 보이지 않았고 일부 그람양성균에 대해서만 활성을 나타내었다. 조박테리오신 용액에 각각의 단백질 분해효소를 처리한 후 박테리오신의 잔존활성을 측정한 결과 분리균이 생산하는 박테리오신은 대부분의 단백질분해효소에 의해 활성이 제거되었다. 그리고 조박테리오신 용액을 열처리를 한 뒤 잔존활성을 측정한 결과 분리균이 생산하는 박테리오신은 100℃에서도 매우 안정하였으며, autoclave를 하여도 50％ 이상의 활성이 유지되었다. 일반적인 박테리오신과 마찬가지로 분리균이 생산하는 박테리오신은 bacteriocidal action을 하는 것으로 관찰되었다.

• Asian-Australasian Journal of Animal Sciences
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• 제18권5호
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• pp.741-746
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• 2005

To investigate the basic information and the possibility of ACE-inhibitory peptides for antihypertension materials, goat's caisin (CN) was hydrolyzed by various proteolytic enzymes and ACE-inhibitory peptides were separated and purified. ACE-inhibition ratios of enzymatic hydrolysates of goat's CN and various characteristics of ACE-inhibitory peptides were determined. ACE-inhibition ratios of goat's CN hydrolysates were shown the highest with 87.84% by pepsin for 48 h. By Sephadex G-25 gel chromatograms, Fraction 3 from goat's CN hydrolysates by pepsin for 48 h was confirmed the highest ACE-inhibition activity. Fraction 3 g and Fraction 3 gh from peptic hydrolysates by RP-HPLC to first and second purification were the highest in ACE-inhibition activity, respectively. The most abundant amino acid was leucine (18.83%) in Fraction 3 gh of ACE-inhibitory peptides after second purification. Amino acid sequence analysis of Fraction 3 gh of ACE-inhibitory peptides was shown that the Ala-Tyr-Phe-Tyr, Pro-Tyr-Tyr and Tyr-Leu. IC$_{50}$ calibrated in peptic hydrolysates at 48 h, Fraction 3, Fraction 3 g and Fraction 3 gh from goat's CN hydrolysates by pepsin for 48 h were 29.89, 3.07, 1.85 and 0.87 g/ml, respectively. Based on the results of this experiment, goat's CN hydrolysates by pepsin were shown to have ACE-inhibitory activity.

• Archives of Plastic Surgery
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• 제34권3호
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• pp.305-313
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• 2007

Purpose: Wound healing is a result of complex processes whose components, such as cells, extracellular matrix, proteolytic enzymes, and their inhibitors receive effects from immune compartments, cytokines, chemokines, and growth factors. Impairment of normal physiologic response to wounding makes nonhealing chronic wounds. Wound infection and exacerbated proteolytic process may induce uncontrolled tissue degradation or exudates formation, which may result in the development of a nonhealing chronic wound. Thus proper management of wound infection and exudates is critical to prevent and treat nonhealing wound. The aim of this study is to evaluate effects of Aquacel AG, silver-containing carboxymethylcellulose dressing on treatment for exudative infected wound. Methods: The study included 31 patients with nonhealing wound. Wound was dressed with Aquacel AG. The effect of dressing was investigated by serial bacterial culture and wound exudates assessment. Each infection and exudates control time was determined and statistically analyzed. Results: Wound infection and exudates were effectively managed using Aquacel AG dressing. Mean infection and exudates control time were $3.4{\pm}1.2$ and $5.7{\pm}1.4$ weeks, respectively. Statistical analysis of the data indicated that infection control time correlated positively to age and exudates control time (p<0.05). Conclusion: There is as yet no ideal dressing for the topical treatment of chronic nonhealing wound. But silver-containing carboxymethylcellulose dressing can be used effectively for exudative, infected nonhealing wound.

• Journal of Animal Science and Technology
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• 제47권1호
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• pp.83-90
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• 2005

ACE-inhibitory peptides derived from goat's whey hydrolyzed by various proteolytic enzymes were separated and purified for antihypertension materials. The highest ACE-inhibitory activity of goat's whey hydrolysates was 85.5 % by pepsin for 72 hrs. Also the highest ACE-inhibitory activity of goat's whey hydrolysates was F-4 by pepsin for 72 hrs by Sephadex G-25 gel chromatograms. F-4e and F-4ed from F-4 by RP-HPLC to first and second purification were the highest in ACE-inhibitory activity, respectively. The most abundant amino acid was leucine(I 8.54 %) in F-4ed of ACE-inhibitory peptides after second purification. Amino acid sequence of F-4ed of ACE-inhibitory peptides showed Leu-Lys-Asp-Tyr-Gly-GlyVal- Ser-Leu and Leu-Gly-Asp-Gly-Ala-Gly- Asp-Val-Ala-Phe. $IC_{50}$ calibrated in peptic hydrolysates(72 hrs), F-4, F-4e and F-4ed from goat's whey hydrolysates by pepsin for 72 hrs were 33.93, 28.75, 11.74 and 1.09 mg/ml, respectively. From the results of this experiment, goat's whey hydrolysate by pepsin was shown to have ACE-inhibitory activity.

• 미생물학회지
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• 제18권4호
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• pp.161-171
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• 1980

A mutant strain having increased productivity of both enzymes, protease and amylase, was obtained from A. flavus KU 153, isolatd from South Korea for its high protease production by successive ultra-violet light irradiation, Two glucoamylases from the mutant strain selected were purified from wheat branculture by successive salting out, followed by dialysis and column chromatography, and their characteristics were compared with those of the wild strain. Glucoamylase production of the mutant selected was increased about 3.3 times compared with the wild strain, and 2.1 times compared with the parental strain, ${\alpha}-amylase$ activity of the mutant selected was about 2 times hugher than that of the wild strain or the parental strain. Protease and cellulase productivities of the muant selected were all alike compared with those of the highly proteolytic mutant, the parental strain. Therefore, it was considered that the back mutation on the protease production did not occurred in the formation process of the glucoamylase producing mutant. Total activities of glucoamylase I and II from the mutant selected were 2.86 and 3.65 times higher compared with those from the wild strain, respectively. Considering the optimal pH-thermal stability and Km-Vmax value of glucoamylase I and II from both strains, wild and mutant, it was deduced that the characteristics of glucoamylase I and II from the wild strain did not altered during the mutation process. Therefore, it was concluded that the selected mutant did not induce the formation of another glucoamylase isozyme, or the changes in the characteristics of the glucoamylase, but induce the productivity of the same glucoamylase I and II by the action of regulatory gene.

• 한국미생물·생명공학회지
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• 제49권1호
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• pp.65-74
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• 2021

Serine proteases are the most versatile proteolytic enzymes with tremendous applications in various industrial processes. This study was designed to investigate the biochemical properties, critical residues, and the catalytic potential of alkaline serine protease using in-silico approaches. The primary sequence was analyzed using ProtParam, SignalP, and Phyre2 tools to investigate biochemical properties, signal peptide, and secondary structure, respectively. The three-dimensional structure of the enzyme was modeled using the MODELLER program present in Discovery Studio followed by Molecular Dynamics simulation using GROMACS 5.0.7 package with CHARMM36m force field. The proteolytic potential was measured by performing docking with casein- and keratin-enriched residues, while the effect of the inhibitor was studied using phenylmethylsulfonyl fluoride, (PMSF) applying GOLDv5.2.2. Molecular weight, instability index, aliphatic index, and isoelectric point for serine protease were 39.53 kDa, 27.79, 82.20 and 8.91, respectively. The best model was selected based on the lowest MOLPDF score (1382.82) and DOPE score (-29984.07). The analysis using ProSA-web revealed a Z-score of -9.7, whereas 88.86% of the residues occupied the most favored region in the Ramachandran plot. Ser327, Asp138, Asn261, and Thr326 were found as critical residues involved in ligand binding and execution of biocatalysis. Our findings suggest that bioengineering of these critical residues may enhance the catalytic potential of this enzyme.

• 미생물학회지
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• 제37권1호
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• pp.1-8
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• 2001

거미는 육식동물로 구강의 소화라는 독특한 방법을 통하여 곤충을 비롯한 작은 동물을 먹이자원으로 이용한다. 거미의 독샘에 함유되어 있는 단백질 분해 효소 환만 아니라 소화관에 존재하는 미생물도 거미의 소화에 중요한 역할을 할 것으로 추정된다. 본 연구에서는 한국산 무당거미(Nephila clavata)의 소화관내 미생물 군집의 분포와 단백질 및 지질 분해능을 확인하고, 거미의 장내 미생물을 분리 .동정하고자 하였다. 한국산 무당거미의 소화관에 존재하는 총 개체수는 거미 18개체를 통합하여 처리하였을 때와 개체별로 처리하였을 때 모두 거미 한 마리당 $10^3-10^5$CFUs 로 매우 유사하였다. 계수된 미생물 중에서 90% 이상이 단백질 또는 지질 분해능을 나타내었다. 그리고 계수된 미생물 중에서 군종별로 1균주씩 순수 분리하였고, 분리된 미생물 중 63.3%가 각각 단백질 또는 지질 분해능을 나타내었고, 이중 50%의 균주는 단백질과 지질 분해능을 동시에 함유하는 것으로 나타났다. 형태적, 생리 .생화학적 방법을 통하여 동정한 결과, 11종류의 그람음성균(Acinetobacter calcoaceticus, A. haemolyticus, Aicaligenes faecalis, Cedecea davisae, C. neteri, Klebsiella pneumoniae, Proteus vulgaris, Pseudomonas fluorescens, Serratia marcescens, Stenotraphamonas maltophilia, Suttonella indologenes)과 11종류의 그람양성균(Bacillus cereus, B. coagulans, B. pasteurii, B. thuringiensis, Cellulomonas flavigena, Corynebacterium martruchotii, Enterococcus durans, E. faecalis, Micrococcus luteus, Staphylococcus huminis, S. sciuri)으로 분류되었다.