• 한국미생물·생명공학회지
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• 제18권5호
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• pp.501-505
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• 1990

토양으로부터 분리한 Streptomyces 속 세균 KIS 13은 thiol 계통 단백질분해효소 활성을 특이적으로 저해하는 저분저량 저해물질을 생성하였다. 저해물질 생성은 세균체성장에 연관된 생성양상이 나타내었다. 배양액으로부토 butanol 추출, silicagel 60 column chromatography, Sephadex LH-2 gel-filtration chromatography, preparative HPLC 등의 과정을 통하여 단백질 분해효소 저해물질을 순수분리하였다. 이 저해물질은 Hammersten casein을 기질로 사용할때, papain에 대하여 non-competitive한 저해양상을 나타내었다.

• 한국식품영양과학회지
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• 제20권1호
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• pp.21-26
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• 1991

the Bacillus subtilis LY-353 which secretes the protease isolated from seafoods. The opti-mum culture condition for production of protease from B. subtilis LY-353 was as follows ; tem-perature 35$^{\circ}C$ pH 7.5 salt concentration 1.0% The purification steps involved ammonium sulfate fractionation DEAE-Sephadex A-50 column chromatography and Sephadex G-100 gel filtration A 7.33 fold purification and 6.55 yield of protease was obtained from culture broth, The optimum pH and temperature for the enzyme action were pH7.5 and 55$^{\circ}C$ respecti-bely.

• KSBB Journal
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• 제16권2호
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• pp.158-162
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• 2001

A halotolerant and extracellular protease-producing yeast was isolated from traditional Meju and identified as a strain of Hansenular polymorpha by investigating its microbiological characteristics. The optimum pH, temperature and NaCl concentration reauired for the growth of Hansenular polymorpha S-9 were found to be pH 6.0, 30$^{\circ}C$ and 0.5 M, respectively. Extracellular protease was produced maximally at 10 U ml(sup)-1 when Hansenular polymorpha S-9 was grown on the medium containing 1.0% beef extract and 0.1 M NaCl for 12 hr at 30$^{\circ}C$. About 13% of the angiotensin-converting enzyme (ACE) inhibitory activity was shown in the hydrolysates which were obtained from the digestion of soybean protein (6 mg) for 6 hr at 30$^{\circ}C$ by the crude enzyme (1 U).

• Journal of Microbiology and Biotechnology
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• 제21권6호
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• pp.627-636
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• 2011

A commercially important alkaline protease, produced by Bacillus sp. RRM1 isolated from the red seaweed Kappaphycus alvarezii (Doty) Doty ex Silva, was first recognized and characterized in the present study. Identification of the isolated bacterium was done using both biochemical characterization as well as 16S rRNA gene sequencing. The bacterial strain, Bacillus sp. RRM1, produced a high level of protease using easily available, inexpensive agricultural residues solid-state fermentation (SSF). Among them, wheat bran was found to be the best substrate. Influences of process parameters such as moistening agents, moisture level, temperature, inoculum concentration, and co-carbon and co-nitrogen sources on the fermentation were also evaluated. Under optimized conditions, maximum protease production (i.e., 2081 U/g) was obtained from wheat bran, which is about 2-fold greater than the initial conditions. The protease enzyme was stable over a temperature range of 30-$60^{\circ}C$ and pH 6-12, with maximum activity at $50^{\circ}C$ and pH 9.0. Whereas the metal ions $Na^+$, $Ca^{2+}$, and $K^+$ enhanced the activity of the enzyme, others such as $Hg^{2+}$, $Cu^{2+}$, $Fe^{2+}$, $Co^{2+}$, and $Zn^{2+}$ had rendered negative effects. The activity of the enzyme was inhibited by EDTA and enhanced by $Cu^{2+}$ ions, thus indicating the nature of the enzyme as a metalloprotease. The enzyme showed extreme stability and activity even in the presence of detergents, surfactants, and organic solvents. Moreover, the present findings opened new vistas in the utilization of wheat bran, a cheap, abundantly available, and effective waste as a substrate for SSF.

• 한국식품위생안전성학회지
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• 제16권1호
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• pp.33-36
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• 2001

V. parahaemolyticus를 균수로 alkaline protease 생산을 위한 배지의 초기 pH, 배양온도 그리고 배양시 진탕속도에 따른 영향을 조사하였다. 배지의 초기 pH에 따른 변화는 pH 7.6에서 60시간, 72시간 배양한 후에는8.lunit, 7.4unit로 활성이 가장 높게 나타났고, pH 7.0, 8.0 그리고 9.0에서는 활성이 크게 낫아졌으며, pH 6.0과 10.0에서는 활성이 전혀 나타나지 않았다. 배앙온도에 따른 변화를 보면 37$^{\circ}C$에서 60시간, 72시간 배양한 후에는 7.3unit, 6.9unit로. 활성이 가장 높게 나타났고, $25^{\circ}C$와 3$0^{\circ}C$에서 배양한 경우에는 2.5unit 전후로 활성이 크게 낮아졌으며, 45$^{\circ}C$에서는 활성이 거의 나타나지 않았다. 배양시 진탕속도에 따는 변화를 보면 250 rpm에서 60시간, 72시간 배양한 후에는 6.87unit, 6.9unit로 활성이 가장 높게 나타났고, 150rpm에서는 활성이 1.5unit 전후로 급격히 낮아졌으며 정치 배양한 경우에는 활성이 거의 나타나지 않았다.

• Animal Bioscience
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• 제37권7호
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• pp.1277-1288
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• 2024

Objective: This study was aimed to investigate the effect of fresh and dried hydrolyzed Cordyceps militaris (CM) mushroom with proteolytic enzymes; bromelain (CMB), flavorzyme (CMF), and mixture of bromelain: flavorzyme (CMBF) on quality properties of spent hen chicken. Methods: Mushroom extract (CME) were combined with three proteolytic enzyme mixtures that had different peptidase activities; stem bromelain (CMB), flavorzyme (CMF), and mixture of stem bromelain:flavorzyme (CMBF) at (1:1). The effect of these hydrolysates was investigated on spent hen breast meat via dipping marination. Results: Hydrolyzation positively alters functional properties of CM protease. in which bromelain hydrolyzed group (CMB) displayed the highest proteolytic activity at 4.57 unit/mL. The antioxidant activity had a significant increment from 5.32% in CME to 61.79% in CMB. A significantly higher emulsion stability index and emulsification activity index compared to CME were another result from hydrolyzation (p<0.05). Texture properties along with the shear force value and myofibrillar fragmentation index were notably improved under CMB and CMBF in fresh condition. Marination with CM mushroom protease that was previously hydrolyzed with enzymes was proven to also increase the nucleotide compounds, indicated by higher adenosine 5'-monophosphate (AMP) and inosine 5'-monophosphate (IMP) in hydrolysate groups (p<0.05). The concentration of both total and insoluble collagen remained unchanged, meaning less effect from CM protease. Conclusion: This study suggested the hydrolyzation of CM protease with bromelain or a mixture of bromelain:flavourzyme to significantly improve functional properties of protease and escalate the taste-related nucleotide compounds and texture profiles from spent hen breast meat.

• 대한바이러스학회지
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• 제29권3호
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• pp.175-182
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• 1999

An attractive target for anti-herpes chemotherapy is the herpes simplex virus 1 (HSV-1) protease encoded by the UL26 gene. HSV-1 protease is essential for DNA packaging and virus maturation. To perform high throughput for potent inhibitors, the efficient production of larger amounts of highly purified enzyme and protease activity assay method must be established. In this report, expression in E. coli and purification of the protease gene of HSV-1 strain F was investigated. The protease gene was cloned pET28, and the nucleotide sequence of protease catalytic domain of HSV-1 compared strain F with other strains (KOS and CL101). In these results the F strain was different in base sequence. However, the amino acid sequence was identifical. The HSV-1 protease was purified with His-tagged affinity column. The analysis of HSV-1 protease activity was performed by high performance liquid chromatography.

• Applied Biological Chemistry
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• 제44권4호
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• pp.246-250
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• 2001

토양시료로부터 높은 활성의 pretense를 생산하는 세균을 수십종 분리하였다. 분리된 세균중에서 protease활성과 성장속도면에서 가장 우수한 균주를 선별하여 WRD-2로 명명하였으며, 형태학적, 생화학적 및 생리학적 특성을 조사한 후 Bacillus sp로 동정되었다. WRD-2조효소액의 protease 최적 활성은 pH 6.0, 온도는 $40^{\circ}C$로 중성 protease임을 알 수 있었고, 온도 $20{\sim}40^{\circ}C$에서 높은 protease활성을 나타내었다. 또한, 초기 pH에 따른 protease 활성에서는 pH 6에서 가장 높은 활성을 나타내었고, 배양배지의 영향은 탄소원으로 maltose 3%, 질소원으로 yeast extract 4%에서 가장 높은 pretense활성을 나타내었다.

• 한국수소및신에너지학회논문집
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• 제28권6호
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• pp.697-703
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• 2017

This study investigated the high-efficient process for bioethanol from barley by various condition. First, higher concentrations of ethanol could be produced without loss of yield by using reducing water consumption. This is because it could prevent to increase viscosity despite reducing water consumption. Second, the ethanol yield could be improved by using reducing particle size of biomass (increase of enzyme reactive surface). Third, The addition of protease could have a considerable effect on yield of fermentation, which provides nutrients to the yeast. This results showed that bioethanol production would provide efficient ethanol production and lower production costs.

• 한국식품과학회지
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• 제32권1호
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• pp.154-160
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• 2000

한국 전통 메주로부터 분리 동정한 Bacillus subtilis PCA20-3이 생산하는 protease의 생산조건과 특성을 조사하였다. Protease의 생산 최적조건을 0.2% soytone, 2% starch, 0.1% $(NH_4)_2SO_4,\;0.2%\;CaCl_2,\;0.01%\;yeast\;extract,\;0.1%\;K_2HPO_4,\;0.1%\;KH_2PO_4$, pH 7.0, 30에서 20시간이었다. 효소의 최적작용 pH와 온도는 6.0-11.0, $55^{\circ}C$였고 pH $6.0{\sim}11.0$의 범위와 $50^{\circ}C$이하에서 안정하였다. 금속이온중 $Fe^{(2+)}$와 $Cu^{(2+)}$에 의해 효소활성이 저해되었다. 2mM의 phenymethanesulfonyl fluoride에 의해 89.2%의 활성이 저하되어, 활성 serine을 가진 serine protease임이 시사되었다. 조효소액의 Km값은 $5.0{\times}10^{(-4)}M,\;V_(max)$값은 $100\;{\mu}g/min$이었으며 bovine serum albumin, isolated soybean protein 보다 casein에 대해 초기 가수분해력이 높은 것으로 나타났다.