• Korean Journal of Food Science and Technology
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• v.8 no.3
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• pp.141-146
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• 1976

1) The purified enzyme was obtained with the specific activity 126.5 u/mg protein (about 45 times of the original activity) and the yield of 4.2%, by means of salting out with ammonium sulfate (0.5 saturation) of the crude enzyme solution, desalting by Sephadex G 25, CM cellulose columm chromatography, concentration by Sephadex G 25, and gel filtration by Sephadex G 75. 2) In the acrylamide gel disc electrophoresis of the purified enzyme, the main band and two obscure ones on the both side of the main band appeared, which indicated that the enzyme was considerably purified compared with its crude enzyme solution, even if it is not referred to as a pure protein.

• BMB Reports
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• v.28 no.5
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• pp.443-450
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• 1995

Cloned staphylococcal peptidoglycan hydrolase was used in determining the physiological characteristics of peptidoglycan hydrolase. This enzyme hydrolyzed the bacterial cell walls and released the N-terminal alanine, but not the reducing groups. This cloned gene product was localized in the cytoplasm of transformed Escherichia coli. Activity gels indicated the enzyme had an Mr of about 54,000, which was consistent with the deduced Mr from sequencing of the cloned gene. The activity bound to CM-cellulose but not DEAE-cellulose resin, indicating it as a basic protein. Enhanced enzyme activity in a low concentration of cations, and inhibited enzyme activity in a solution with dissolved phospholipids, suggested that the activity and the availability of this basic protein may be regulated between negatively charged and positively charged cellular molecules. The activity against boiled crude cell wall was much greater than against purifed cell wall, suggesting protein associated with crude cell wall may aid in the binding of the peptidoglycan hydrolase The cloned peptidoglycan hydrolase showed positive activity on whole cells of some lysostaphin-resistant coagulase-negative staphylococci. The cloned enzyme may be an alternative for lysostaphin for lysis of staphylococci.

• Journal of Pharmaceutical Investigation
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• v.18 no.3
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• pp.125-131
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• 1988

This study was undertaken to investigate the proteolytic enzyme from Neungee mushroom [Sarcodon aspratus (Berk) S. Ito]. The proteolytic activity of Neungee was higher than other several edible mushrooms under various pHs. The potency of proteolytic enzyme of Neungee was same as the digestive drugs containing protease. So the proteolytic activity of the enzyme was increased in neutral or weak alkaline pH, whose characteristics would be alkaline protease. The specific activity of the purified enzyme obtained by using Tris acryl CM-cellulose ion exchange increased 20 times as compared with that of the crude extract. The proteolytic enzyme was stable at room temperature, but decomposition was fast when incubated at higher temperature more than $40^{\circ}C$. The half life of the enzyme was longest in neutral pH and rate constant was increased in acidic or alkaline solution.

• Microbiology and Biotechnology Letters
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• v.20 no.6
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• pp.681-686
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• 1992

In the conversion of D.L-2-amino-$\Delta^2$-thiazoline-4-carboxylic acid(D,L-ATC) to L-cysteine using Pseudomonas sp. CU6. the effects of surfactants on whole cells and the stabilities of cellfree enzyme solution and continuous reactor packed with immobilized whole cells were investigated. The enzymatic reaction was little accomplished by whole cells without adding surfactants, whereas it was well carried out with SDS or Triton X-loo comparable to the case using cell-free enzyme solution. Enzyme activity of the cell-free solution was lost by 50% after 7 hours of storage at $30^{\circ}C$, but not at all under an anaerobic condition by sparging nitrogen gas. On the other hand. effect of nitrogen gas did not appear in a continuous reactor using immobilized whole cells, and hydroxylamine, an inhibitor of L-cysteine desulfhydrase, lowered the enzyme stability.

• Korean Journal Plant Pathology
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• v.3 no.2
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• pp.124-130
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• 1987

The optimum conditions for protoplast formation and regeneration from Pyricularia oryzae Cav. were selected as follows. As a basic solution, 0.02M potassium phosphate buffer solution plus 0.6M KCl adjusted to pH 5.2 with 1N HCl was used. A mixture of enzyme combinations with 20mg Cellulase R-l0/ml, 5mg Macerozyme R-l0/ml and l0mg Driselase/ml used as a lytic enzyme showed better lytie effect than any single enzyme treatment for protoplast formation. Two-day-old mycelia of P. oryzae grown in the mixture of three lytie enzyme solution at $30^{\circ}C$ for 3 hr showed best condition for protoplasts formation. For regeneration from the protoplasts of P. oryzae, potato dextrose agar containing 0.02M potassium phosphate plus 0.6M KCl used as a stabilizer was best for regeneration medium.

• Korean Journal of Microbiology
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• v.36 no.1
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• pp.69-73
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• 2000

Arthrobacter sp. A-6 was cultivated and DFA III(di-fructofuranose dianhydride) was produced with inulin fructotransferase from the chicory root. The specific growth rate, yield of cell mass and yield of enzyme from the culture in variable chicory root extracts were studied and the results compared. Standard inulin solution(10%) was treated with the crude enzyme solution of inulin fructotransferase from the cell culture, 1.14mg/ml of DFA III was produced. The enzyme reactions were processed with various preparations of chicory root extracts in the same conditions. The highest yield of DFA III production(2.29 mg/ml) was obtained from the chicory roots without washing or extraction. The yield of DFA III from the washed chicory roots without extraction was at lowest(0.44 mg/ml). The production process of inulin fructotransferase and DFA III from the chicory root without prewashing or extraction steps were more efficient.

• Journal of the Korean Wood Science and Technology
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• v.24 no.1
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• pp.48-53
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• 1996

Brown rot fungus, Tyromyces palustris, has been reported to cause the loss of strength accelerated by oxalate, a non-enzymatic low molecular weight acid, with minute weight loss of decaying wood in early stage. The production of oxalate in relation to wood decaying and the presence of oxaloacetase. an oxalate producing enzyme, were identified during the process. Tyromyces palustris produced the largest amount of oxalate among brown rot fungi. In order to find out the cleavage of pulp fiber, we submerged pulp fiber in oxalate solution and the results showed that the number of short pulp fiber was highly increased, compared with control solution. The pH of decaying wood was decreased to 1.77 which was close to that of saturated oxalate solution, pH 1.2, Thus, the oxalate was thought to be accumulated in the decaying wood, The oxaloacetase which accelerates production of oxalate was derived from fungus, and the production of oxalate by the enzyme was determined by using on UV/Vis spectrophotometer. Therefore, the oxalate was found to be produced by oxaloacetase during decay. The oxalate may cause the acid-hydrolysis of cellulose and hemicellulose. The oxalate was thought to reduce the degree of polymerization and increase the enzyme activity, which resulted in rapid loss of strength in early stage-an identical feature of brown rot fungus.

• Archives of Pharmacal Research
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• v.3 no.1
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• pp.7-12
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• 1980

Ampliciline was synthesized from 6-amino-pencillanic acid (6-APA) and D-.alpha. phenylglycine methyl ester by using amplicilin synthesizing enzyme from Peudomonas melanogenum (IAM 1655). The whole cell enzyme was immobilized by entrapping it in the polyacrylamide gel lattices. The polymer used in the enzyme entrapment was made from 150 mg per ml of acrylamide monomer and 8 mg per ml of N, N'-methylenebisacrylamide. About 200 mg/whole cell enzyme was mixed in the polymer for entrapment. The maximal activity retention after immobilization was 56%. The optimal pH values for the whole cell enzyme and the immobilized whole cell enzyme were 6.0 and 5.9, respectively. The optimal temperature for the enzyme activity were the same for both type of preparations. The enzyme stabilities against pH and heat increased for immobilized whole cell enzyme. Immobilized cell was more stable especially in the acidic condition while both type were found to be very suceptible to thermal inactivation at a temperature above 4.deg.C. The kinetic constants obtained from Lineweaver-Burk plot based on two substate reaction mechanism showed somewhat higher value for immobilized whole cell enzyme as compared to the whole cell enzyme : the Km value for 6-APA were 7.0 mM and 12.5 mM while Km values for phenylglycine methyl ester were 4.5 mM and 8.2 mM, respectively. Using the immobilized whole cell enzyme packed in a column reactor, the productivity of ampiciline was studied by varying the flow rate of substrate solution. At the space velocity, SV, 0.14 hr$^{-1}$ the conversion was 45%. Operational stability found in terms of half life was 30 hr at SV = 0.2 hr.

• Fibers and Polymers
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• v.6 no.3
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• pp.181-185
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• 2005

Silk fibroin (SF) nanofibers were prepared by electrospinning and their application as an enzyme immobilization support was attempted. By varying the concentration of SF dope solution the diameter of SF nanofiber was controlled. The SF nanofiber web had high capacity of enzyme loading, which reached to $5.6\;wt\%$. The activity of immobilized a-chymotrypsin (CT) on SF nanofiber was 8 times higher than that on silk fiber and it increased as the fiber diameter decreased. Sample SF8 (ca. 205 nm fiber diameter) has excellent stability at $25^{\circ}C$ by retaining more than $90\%$ of initial activity after 24 hours, while sample SF11 (ca. 320 nm fiber diameter) shows higher stability in ethanol, retaining more than $45\%$ of initial activity. The formation of multipoint attachment between enzyme and support might increase the stability of enzyme. From these results, it is expected that the electrospun SF nanofibers can be used as an excellent support for enzyme immobilization.

• 한국생물공학회:학술대회논문집
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• 2000.11a
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• pp.567-570
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• 2000

The condition of immobilization of the partially purified levan fructotransferase and the properties of the immobilized enzyme was investigated. Levan fructotransferase was immobilized on ${\kappa}\;-carrageenan$ beads by entrapment method. The optimal ${\kappa}\;-carrageenan$ concentration was obtained 2%(w/v) (or the matrix. At that time, immobilized enzymes(0.81 units) have relative low activity compare with soluble enzyme(7.7 units). To immobilized and soluble enzyme, optimal activity temperature and pH were measured $55^{\circ}C$, 6.0 in sodium phosphate buffer 20mM solution. If crosslinking agent was added, proper concentration was 0.5%(v/v). At $37^{\circ}C$, immobilized and soluble enzyme converted levan to oligofructose and DFA IV, and the conversion ratio was 32% and 61% at 60 hr.