• Food Engineering Progress
• /
• v.15 no.3
• /
• pp.214-220
• /
• 2011

$\beta$-Glucosidase enzyme reaction under high hydrostatic pressure was investigated in terms of physical chemistry. A model substrate (p-nitrophenyl-${\beta}$-D-glucopyranoside(pNPG)) was used, and the pressure effects on the enzymatic hydrolysis (pNPG${\rightarrow}$pNP) at 25 MPa, 50 MPa, 75 MPa, and 100 MPa were analyzed. Two parts of the reaction such as kinetic and equilibrium stages were considered for mathematical modelling, and their physicochemical parameters such as forward and inverse reaction constants, equilibrium constant, volume change by pressure, etc. were mathematically modeled. The product concentration increased with pressure, and the two stages of reaction were observed. Prediction models were derived to numerically compute the product concentrations according to reaction time over kinetic to equilibrium stages under high pressure condition. Conclusively, the $\beta$-Glucosidase enzyme reaction could be activated by pressurization within 100 MPa, and the developed models were very successful in their prediction.

• Journal of Life Science
• /
• v.14 no.4
• /
• pp.608-613
• /
• 2004

Pyrimidine nucleotide N-ribosidase (pyrimidine 5'-nucleotide phosphoribo (deoxyribo) hydrolase/pyrimidine 5'-nucleoude nucleosidase, EC 3.2.2.10) directly catalyzes pyrimidine 5'-nucleotide to pyrimidine base and ribose (deoxyribo) 5-phosphate. In order to clarify the best nutritional conditions for the growth and the pyrimidine nucleotide N-ribosidase production of Pseudomonas oleovorans ATCC 8062 the effects of various nutrients such as different carbon and nitrogen sources were studied. For the both the growth and the enzyme production, 2% fumarate, 1.5% peptone, 5% corn steep liquor (CSL) and 1% ammonium chloride were excellent carbon and nitrogen sources, respectively. Optimum pH, temperature, and cultivation time for the enzyme production were 7.0, $28^{\circ}C$, and 48 h, respectively. The pyrimidine nucleotide N-ribosidase of P. oleovorans ATCC 8062 was not induced by UMP and its derivatives, and was constitutive enzyme.

• Korean Journal of Microbiology
• /
• v.37 no.1
• /
• pp.15-20
• /
• 2001

In this study, we have isolated bacterial cell wall lytic enzyme in the culture supernatant of Aspergillus sp. HCLF-4. This hydrolase showed cell wall lytic activity against Anabaena cylindrica. The extracellular enzyme was produced by Aspergillus sp. HCLF-4 when it was grown in a PDB media containing 0.05% heat killed Micrococcus luteus cells. The molecular weight of lytic enzyme was about 14.3 kDa. The optimal pH and temperature for the activity of this enzyme were 3.0~4.0 and $30^{\circ}C$, respectively. This hydrolase activity was reduced by $Na^{+}$, $Li^{+}$, $Ca^{2+}$, $Cu^{2+}$, $Fe^{3+}$, EDTA, and PMSF, whereas it was increased by $Mg^{2+}$, $Mn^{2+}$>. The enzyme has N-acetylmuramyl-L-amidase or endopeptidase activity.

• Proceedings of the Korean Society of Applied Pharmacology
• /
• 1994.04a
• /
• pp.274-274
• /
• 1994

본 연구에서는 토끼신장 근위세뇨관 일차배양세포에서 브라디키닌의 생리작용이 phospholipase D (PLD)에 의해 매개되는지를 살펴 보기위해 PLD 효소반응의 특이한 성질인 transphosphatidylation 반응의 생성물인 phosphatidylethanol (PEth) 의 세포내 양을 측정함으로 PLD 효소의 관련성을 규명할 수 있었다. 시간경과에 따른 phosphatidic acid (PA) 및 diacylglycerol (DAG) 의 생성을 살펴본 결과 PA가 DAG보다 먼저 생성되어 최고치 (30초)에 도달하였고 DAG는 1분이후부터 5분까지 서서히 생성되는 양상을 나타내었다. 또한 0.5에서 5％까지의 에탄올 존재하에 PA 및 PE소 생성량을 비교해본 결과 에탄올량이 증가함에 따라 PA는 감소하는 반민 PEth 의 생성은 계속 증가하였다. 한편 브라디키닌 농도 변화 실험에서는 브라디키닌농도가 증가함에 따라 PA 및 PEth 둘다 생성이 증가되었다. 이러한 결과로부터 토끼신장 근위세뇨관 세포막에 존재하는 브라디키닌수용체는 브라디키닌에 의해 activation 시 PLD를 직접적으로 활성화시켜 그들의 작용을 세포내로 전달한다는 사실을 알 수 있었다. 또한 PLD 효소활성의 activator로 수용체효능 제외에 칼슘이온, protein kinase C (PKC) 등이 몇몇 다른 실험에 의해 밝혀져 있고, G protein 역시 PLD 효소 활성을 조절하는 역할이 있음이 알려졌다. calcium ionophore 및 칼슘채널길항제인 verapamil을 이용한 실험에서 우리는 브라디키닌의 PLD 활성화는 칼슘이온에 의존적인 경로 및 비의존적인 경로가 같이 존재함을 알수 있었다. 또한 브라디키닌의 PLD 활성화기전이 PKC 의존적인지를 살펴보기위해 PKC activator(PMA) 및 inhibitor (staurosporine)를 이용한 실험에서 브라디키닌은 신장세포에서 PKC를 통하여 PLD를 활성화시킴으로 신호전달을 하는 것으로 추측되었다. 마지막으로 가수분해안되는 G protein 유도체인 GTPrS 및 G protein 활성물질 NaF, 백일해독소등을 이용한 실험에서 G protein 의 PLD 조절활성을 확인할 수 있었다.

• Korean Journal of Soil Science and Fertilizer
• /
• v.39 no.6
• /
• pp.386-391
• /
• 2006

This study was conducted to evaluate the indole acetic acid (IAA) formation in soils as a biological indicator for the health of rice paddy soils with control, nitrogen sole, chemical fertilizer (NPK), and chemical fertilizer plus compost (CNPK) plots. There was a positive relationship between colorimetric method and high performance liquid chromatography for IAA in soils determined, and the values were similar between two methods, as $0.83{\sim}1.23{\mu}g\;IAA\;g^{-1}h^{-1}$ in colorimetric method, $0.80{\sim}1.29{\mu}g\;IAA\;g^{-1}h^{-1}$ in HPLC method. Numbers of dehydrogenase-producing bacteria and the IAA production in soils were high in NPK and CNPK plots comparing with control and nitrogen sole plots. Also there was high correlation between numbers of dehydrogenase-producing bacteria and IAA production in soils.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.21 no.2
• /
• pp.195-200
• /
• 1992

Rhizopus oryzae CJ-2114 was selected for its strong polygalacturonase activity among various strains of mold found in soil. The optimum pH for the enzyme activity was 4.0 and optimum temperature was 4$0^{\circ}C$. The activation energy for the polygalacturonase was calculated by Arrhenius equation was 2.048㎉/㏖. The reaction of this enzyme followed typical Michaelis-Menten kinetics with the Km value of 54.05mM with the $V_{max}$ of 13.9m mole/min. The enzyme is relatively stable in acidic condition. The activity of polygalactur-onase was inhibited completely by C $u^{2+}$, P $b^{2+}$ and Z $n^{2+}$, $_Mn^{2+}$ at concentration of 1 mM. The enzyme can be inactivated by the treatment with maleic anhydride and iodine. The results indicate the possible involvement of histidine at active site. When polygalacturonase from Rhizopus oryzae CJ-2114 was reacted with poly-galacturonic acid as a substrate mono-, di-, and oligogalacturonic acid were produced at early and mono-, digalacturonic acid produced at late incubation time. time.

• Microbiology and Biotechnology Letters
• /
• v.17 no.5
• /
• pp.461-467
• /
• 1989

The regulation of the synthesis of alcohol-oxidase (E.C.1.1.3.13.) was investigated in the methanol-utilizing yeasts during growth on different carbon sources. For this experiment, Hansenula polymoypha CBS 4132, Hansenula polymoypha CBM 11 and Hansenula polymoypha Cooney were cultured in mineral salt medium by changing its carbon sources. The production of alcohol-oxidase was varied by the carbon sources. For exmaple, alcohol-oxidase was undetectable: in all strains submitted to the test in the medium with glucose, but its production was rapidly increased when the carbon source was changed from glucose to methanol after 30 hrs of incubation. Moreover, this enzyme was not synthesized during growth on the primary aliphatic alcohols alone (ethanol, propanol, butanol or pentanol) or on the mixed substrates (0.5% methanol + 0.5% primary aliphatic alcohols). When cells were grown on the various carbon sources (glucose, xylose, lactose, glycerol, galactose, saccharose, sorbose, lactic acid or acetic acid), the alcohol-oxidase was about one-tenth of the activity found in cells grown on methanol alone. These carbon sources together with methanol yielded far better synthesis of alcohol-oxidase than in case of carbon sources alone. Especially, the alcohol-oxidase activity of the cells grown on lactose or lactic acid together with methanol was far better or similar than that of cells grown on methanol alone. The synthetic activity of alcohol-oxidase of Hansenula polymoypha CBS 4132 was the strongest among the three strains tested in every respect.

• Korean Journal of Fisheries and Aquatic Sciences
• /
• v.29 no.3
• /
• pp.309-319
• /
• 1996

Properties as related to the utilization of the crude proteases extracted from the muscle and viscera of fish (2 dark fleshed lish; anchovy, Engraulis japonica, and gizzard-shad, Clupanoda punctatus; 2 white fleshed fish; seabass, Lateolabrax japonicus, and sole, Pleuronichthys cornutus) were studied. Proteolytic activity of the muscle protease was slightly inhibited with the increase of sodium chloride concentration and it was apparent against the yellowtail myofibrillar protein than casein substrate. Proteolytic activities of the seabass and sole visceral crude protease were inhibited to 50 to $60\%\;by\;25\%$ of sodium chloride, but those of anchovy and gizzard-shad viscera crude enzymes were not influenced by sodium chloride. The vacuum freeze-dried crude protease and glycerol-mixed crude pretense of gizzard-shad and seabass muscles were almost lost their activities on the 16th week of storage, while those from the viscera of the fish were relatively stable. Degradation of the yellowtail myofibrillar protein by the anchovy muscle and viscera crude pretenses rapidly proceeded in the beginning of the reaction and the degraded products were mainly distributed in the range of 6 to 15 kDa electrophoretically.

• Microbiology and Biotechnology Letters
• /
• v.8 no.4
• /
• pp.229-235
• /
• 1980

Pectin transeliminase (PTE) was produced by Saccharomyces cerevisiae, Schizosaccharomyces pombe 4683, and Aspergillus niger in the media containing 2% pectin and examined for its characteristics. The Production of the enzyme was higher by Asp. niger than by the two yeast strains, showing that the PTE activity was proportional to reducing power. The enzyme was proved to reduce pectin and produce 4, 5- unsaturated galacturonic acid. The optimum activity of the PTE was found to be at pH 6.0 and $50^{\circ}C$. The activities of these enzyme were stable below $50^{\circ}C$ but decreased at the higher temperature. Substrate inhibition of the PTE activities was appeared at high concentrations of pectin. Those PTE activities were increased under 0.6M of KCI and NaCI, but that maximal activities at the concentration of 0.2M MgC $l_2$.

• Applied Biological Chemistry
• /
• v.36 no.6
• /
• pp.539-546
• /
• 1993

This paper describes the purification and partial characteristics of aminopeptidase from Microccus sp. LL3 to utilize the microorganism as a potential agent for industrial application for the purpose of shortening ripening period of cheddar cheese. The optimal temperature and pH for enzyme activity were $35^{\circ}C$ and 7.0, respectively for L-leucine-p-nitroanilide as substrate. The enzyme remained stable for 10 minutes up to $50^{\circ}C$. The activity of aminopeptidase was stimulated by $Mg^{++}$ ion but strongly inhibited by $Hg^{++}$, metal complexing reagents, ethylenediaminetetraacetate (EDTA) and 1,10-phenanthroline. The enzyme was thought to be metallopeptidase. This enzyme had a broad substrate specificity, but was inactive on peptide with arginine as N-terminal amino acid. An intracellular aminopeptidase from Micrococcu sp. LL3 was purified by chromatography on DEAE-Sephacel and filtration on Sepacryl S-300. The enzyme has a molecular weight of 43,500.