• Journal of Microbiology and Biotechnology
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• 제20권2호
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• pp.332-339
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• 2010

The lipase from Mucor racemosus NRRL 3631 was partially purified by fractional precipitation using 60% ammonium sulfate, which resulted in a 8.33-fold purification. The partially purified lipase was then immobilized using different immobilization techniques: physical adsorption, ionic binding, and entrapment. Entrapment in a 4% agar proved to be the most suitable technique (82% yield), as the immobilized lipase was more stable at acidic and alkaline pHs than the free enzyme, plus 100% of the original activity was retained owing to the thermal stability of the immobilized enzyme after heat treatment for 60 min at $45^{\circ}C$. The calculated half-lives (472.5, 433.12, and 268.5 min at 50, 55, and $60^{\circ}C$, respectively) and the activation energy (9.85 kcal/mol) for the immobilized enzyme were higher than those for the free enzyme. Under the selected conditions, the immobilized enzyme had a higher $K_m$ (11.11 mM) and lower $V_{max}$ (105.26 U/mg protein) when compared with the free enzyme (8.33 mM and 125.0 U/mg protein, respectively). The operational stability of the biocatalyst was tested for both the hydrolysis of triglycerides and esterification of fatty acids with glycerol. After 4 cycles, the immobilized lipase retained approximately 50% and 80% of its original activity in the hydrolysis and esterification reactions, respectively.

• 식품보건융합연구
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• 제6권6호
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• pp.1-7
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• 2020

We compared the fermentation of 0 to 4 weeks by manufacturing a rapid low salt-fermented seahorse with a commercial Protamex added to the functional food, Hippocampus abdominalis. We studied amino acid composition, content and major amino acids related to flavor during the fermentation process of salt-fermented seahorse. In the enzyme-free group, it showed little change in the content of non-protein nitrogenous compounds, the content of amino acids and degree of hydrolysis. The Protamex enzyme treatment group was rapidly hydrolyzed in one week of ripening, resulting in increased non-protein nitrogenous compounds content, amino acid content and degree of hydrolysis, and minimal changes in the four weeks. The total amino acid contents ratio showed the highest content of glutamic acid in the enzyme additive group, glycine, alanine, which indicates sweet taste, and serine, the content of glycine, alanine, serine, and lysine, indicating sweet taste, has increased significantly over the enzyme-free group. Twenty species of free amino acid in the four-week of salt-fermented seahorse were detected. It detected 43.0% (6 species) in the enzyme-free group and 63.96% (7 species) in the enzyme additive group.

• 디지털융복합연구
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• 제14권1호
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• pp.253-262
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• 2016

연산오계는 오래전부터 건강기능 증진 및 치료 효능이 높은 것으로 알려져 왔다. 최근 건강 기능식품 소재로 기능성 펩타이드 효능이 알려짐에 따라, 연산오계 내장 부산물로 부터 올리고 펩타이드 최적 생산 공정 및 생성물 특성에 대하여 연구를 수행하였다. 최적 효소가수 분해 공정 표면반응 분석을 이용하여 수행하였다. 최적 공정 조건을 확립하기 위해서 온도 (40, 50, $60^{\circ}C$), pH (pH 6.0, 7.0, 8.0), 효소 (1, 2, 3%) 범위에서 수행을 하였다. 생성물에 대한 가수분해도, 아미노산, 분자량 분포를 분석하였다. 효소 가수분해 최적 온도는 $58^{\circ}C$, pH 7.5, 효소의 농도는 3% 이었다. 최적 조건에서 2 시간 효소 가수분해를 한 결과 75-80% 이었다. 구성 아미노산의 총 함량은 386.15 mg/100 g 이었고 유리 아미노산 총량은 155.26 mg/100 g. 분자량를 Maldi-TOF 으로 분석을 한 결과 90% 이상이 300-1,000 Da 분포를 보여주었다.

• Food Science and Biotechnology
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• 제18권2호
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• pp.570-573
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• 2009

The debranching enzyme of Nostoc punctiforme (NPDE) is a novel enzyme that catalyzes the hydrolysis of $\alpha$-1,6-glycosidic linkages in starch, followed by the sequential hydrolysis of $\alpha$-1,4-glycosidic linkages. The debranching activity of NPDE is highly specific for branched chains with a degree of polymerization (DP)>8. Moreover, the rate of hydrolysis of $\alpha$-1,4-linkages by NPDE is greatly enhanced for maltooligosaccharides (MOs) with a DP>8. An analysis of reaction mixtures containing various starches revealed the accumulation of maltooctaose (G8) with glucose and maltose. Based on the novel enzymatic properties of NPDE, an MO mixture containing more than 60% G8 with yield of 18 g G8 for 100 g starch was prepared by the reaction of NPDE with soluble starch, followed by ethanol precipitation and gel permeation chromatography (GPC). The yield of the G8-rich mixture was significantly improved by the addition of isoamylase. In summary, a 4-step process for the production of a G8-rich mixture was developed involving the enzymatic hydrolysis of starch by NPDE.

• 한국임상약학회지
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• 제8권1호
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• pp.29-34
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• 1998

Murine CD38 is a 42 kDa type II glycoprotein expressed on cell surface of both B and T lymphocytes. CD38 is a multifunctional enzyme that catalyzes the formation and hydrolysis of cyclic adenosine diphosphoribose (cADPR): ADP-ribosyl cyclase activity of CD38 catalyzes the formation of cADPR from NAD and cADPR hydrolase activity of CD38 catalyzes the hydrolysis of cADPR to ADP-ribose (ADPR). And also, CD38 has the catalytic activity of NAD glycohydrolase (NADase) which catalyzes the hydrolysis of catalyzes the formation and hydrolysis of cyclic adenosine diphosphoribose (cADPR): ADP-ribosyl cyclase activity of CD38 catalyzes the formation of cADPR from NAD to ADPR. In this study, we attempted to purify CD38 from mouse lymphocytes by using the immobilized anti-CD38 monoclonal antibody. The single step immuno-affinity column chromatography resulted in homogeneous purification, showing a single protein of 42 kDa on a SDS polyacrylamide gel. We have investigated the effects of various inhibitors on the enzyme activities of the purified CD38. Cibacron blue (0.5 mM) inhibited all three enzyme activities of CD38, NADase, ADP-ribosyl cyclase and cADPR hydrolase activities. ADPR (2 mM) showed inhibitory effect on both cADPR hydrolase activity and NADase, but not on ADP-ribosyl cyclase activity. However, ATP (2 mM) inhibited only cADPR hydrolase activity. $Zn^{2+}$ (1 mM) showed similar inhibitory effect as that of ADPR, but activated cyclase activity These results suggest that CD38 has three different catalytic activity domains which might be differentially regulated by their specific inhibitors.

• Journal of Microbiology and Biotechnology
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• 제23권8호
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• pp.1133-1139
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• 2013

Enzymatic decomposition of gelatin layers on used X-ray films and repeated utilization of the enzyme for potential application in silver recovery were investigated using keratinolytic serine proteases from Purpureocillium lilacinum LPS # 876. At pH 9.0, the enzymatic reaction was enhanced by the increase of enzyme concentration or by the increase of the temperature up to $60^{\circ}C$. Under the conditions of 6.9 U/ml, $60^{\circ}C$, and pH 9.0, hydrolysis of the gelatin layers and the resulting release of silver particles were achieved within 6 min. The protective effect of polyols against thermal denaturation was investigated. The presence of glycerol and propylene glycol increased enzyme stability. When the reusability of the enzyme for gelatin hydrolysis was tested, it could be seen that it could be effectively reused for more cycles when glycerol was added, compared with the enzyme without protective agents. The results of these repeated treatments suggested that a continuous process of recycling silver from used X-ray is feasible. Keeping in mind that recycling is (at the present time) needed and imperative, it can be remarked that, in this research, three wastes were successfully used: hair waste in order to produce serine proteases; glycerol in order to enhance enzyme thermal stability; and used X-ray films in order to recover silver and PET films.

• 대한화학회지
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• 제22권3호
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• pp.164-172
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• 1978

상온에서 카르복시펩티다제 A에 의한 에스테르 기질의 가수분해 반응을 여러가지 외부 시약의 존재하에 행하였다. 산무수물 형태의 아실-효소중간체가 외부시약에 의해 공격받는다면 아실 부분에 포획된 생성물이나 효소부분에 포획된 생성물이 형성될 것이다. 반응생성물의 분광도와 효소활성도의 변화를 조사한 결과 포획반응의 생성물은 검출되지 않았다. 또한, O-(o-hydroxyphenylacetyl)-L-${\beta}$-phenyllactate가수분해에 대한 효소 반응속도변수를 측정하였다. 이 기질의 o-히드록시기가 분자내 포획기로서 작용하여 산무수물형태의 아실-효소중간체를 공격하여 20coumaranone이 형성되었나를 조사하였으나 분자내 포획반응이 일어났다는 증거는 얻지 못하였다. 이러한 중간체 포획반응의 실패는 포획용 시약이 아실-효소 중간체의 무수산기에 접근할때 입체적 방해를 받거나 중간체의 가수분해 단계도 효소에 의해 촉매되기 때문이라고 생각된다.

• 한국식품과학회지
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• 제26권5호
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• pp.484-489
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• 1994

두유 및 두부제조시 부생되는 비지를 효과적으로 이용하기 위한 방안으로 Aspergillus niger CF-34로부터 얻은 효소를 비지에 처리하여 가용화를 시도하였다. 비지가용화를 위한 본 연구에서 비지에 작용하는 복합효소액의 최적반응조건을 조사하였으며, 복합효소액 처리시 두유비지내 불용성 단백질의 회수율 및 고형분 용해도와 비지의 효소가수분해 특성을 검토하였고, 그 결과는 다음과 같다. 복합효소액의 사용량은 비지 고형분의 약 50%를 가용화하는 2.5%(% of total solid)를 최적 복합효소액의 농도로 선정하였다. 효소작용시간에 따른 비지의 단백질 회수율 및 고형분의 용해도는 효소반응후 3시간에서 각각 약 62%, 50%로 상대적으로 높은 값을 나타내었다. 비지를 alkali 처리(0.1% NaOH)로 입자구조를 변형시킴으로서 효소가수분해율이 증가되었고, 가수분해 시간도 단축됨에 따라 비지 가수분해의 효과적인 전처리방법임을 확인할 수 있었다.

• 한국식품영양학회지
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• 제11권2호
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• pp.228-236
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• 1998

달걀의 Allergenicity를 감소시킬 수 있는 방안을 강구하고자 물리적 처리, 축합 인산염 처리 및 효소처리를 하여 Guinea pig를 이용한 Passive Cutaneous Anaphylaxis(PCA) inhibition 실험과 Non Proteic Nitrogen(NPN)정량을 통한 가수분해율의 측정결과 다음과 같은 결론을 얻었다. 달걀의 allergenicity는 가열에 의해 감소하였으며 가열시간이 길수록 단백질 가수분해율 및 PCA inhibition을 증가 시켰다. Ultraviolet 조사와 Microwave 조사는 단백 가수분해율과 PCA inhibition을 증가 시켜서 allergenicity를 저하시켰으며 ultraviolet이 저해효과가 더 컸으며 부화 달걀은 allergenicity를 감소시키지 않은 것으로 나타났다. 효소처리는 단백질의 가수분해율 및 PCA inhibition을 증가 시키며 allergenicity를 현저히 감소시켰으며 alcalase의 재해효과가 더 컸다. Polyphosphate의 참가는 단백질의 가수분해는 유도하지 않았으나 PCA inhibition을 증가 시키며 allergenicity를 감소시켰다. Allergenicity를 감소시키기 위한 처리를 한 달걀 gel의 주사전자현미경 사진은 효소처리 시 표면이 밝게 나타나서 단백질이 분해되었음을 알 수 있었고 neutrase가 alcalase보다 밝게 나타났으나 반응시간의 증가에 따라 모든 효소 표면이 밝게 나타났다. Instron에서 달걀 gel의 경도를 측정한 결과 효소와의 반응시간이 길수록 경도가 감소하는 경향을 보였다.

• 한국수산과학회지
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• 제44권2호
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• pp.99-103
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• 2011

This study was performed to determine the optimal hydrolysis conditions for the production of a flavoring from the precipitation of snow crab cooker effluent (PSCCE) with commercial proteases. Based on cost-per-enzyme activity and sensory evaluations, Flavourzyme$^{(R)}$ 500 MG plus Protamex$^{(R)}$ (1:1 ratio, w/w) were selected as suitable enzymes. Three independent variables consisting of the substrate concentration (S), enzyme-to-substrate ratio (E/S), and hydrolysis time (T) were examined using response surface methodology (RSM). A model equation obtained from RSM was used to predict the degree of hydrolysis (DH) as follows: % DH = 52.285 - 6.371[S] + 5.469[E/S] + 7.599[T] - $5.818[S]^2$ - $5.633[E/S]^2$ - $6.528[T]^2$ - 3.265[E/S][S] - 5.415[T][S] + 4.315[T][E/S]. From the ridge analysis, the conditions favoring the highest degree of hydrolysis were pH 7.45, $55^{\circ}C$, a S of 21.82%, an E/S of 0.50%, and a T of 3.74 h.