• Title/Summary/Keyword: protease

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Studies on the Production of Protease by Aspergillus oryzae KC-15 and Characteristics of the Enzymes (Aspergillus oryzae KC-15에 의한 protease의 생산 및 그 효소의 특징에 관한 연구)

  • 이미자;정만재
    • Microbiology and Biotechnology Letters
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    • v.8 no.2
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    • pp.77-85
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    • 1980
  • This experiment was conducted to investigate the conditions for production and the characteristics of pretenses. Aspergillus oryzae KC-15, which is selected as a superior strain for the production of the protease, was used in this study. The results obtained were as follows: 1. The optimum culture time for the production of acid, neutral and alkaline protease on wheat bran medium were about 48, 48 and 72hr, respectively. The protease-produced by the strain were mainly alkaline and neutral one, but the production of acid protease was feeble extremely. 2. The addition of NaH$_2$PO$_4$, Na$_2$HPO$_4$, glucose, rice powder and Na-glutamate respectively to wheat bran media were effective for the production of alkaline and neutral protease, and the addition of (NH$_4$)$_2$HPO$_4$, glucose and rice powder respectively were effective for the production of acid protease. 3. Characteristics of professes(equation omitted) 4. As a heat resistance agent, NaH$_2$PO$_4$was the most effective one. The optimum amount of NaH$_2$PO$_4$was 10mg for alkaline and neutral protease, and 5mg for acid protease. 5. The heat resistance of the Protease by NaH$_2$PO$_4$was not recognized mostly above 6$0^{\circ}C$. 6. After the treatment of enzyme solution with 10mg of NaH$_2$PO$_4$for 30 minutes at 55$^{\circ}C$, the residual activities measured for alkaline, neutral and acid protease were 58, 57 and 55% respectively.

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Optimization of Submerged Culture Conditions for Protease Production and Its Enzymatic Properties (Protease 생산을 위한 최적 배양조건 및 생산된 Protease의 특성)

  • Cho, Hee-Yeon;Cho, Nam-Seok
    • Journal of the Korean Wood Science and Technology
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    • v.32 no.5
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    • pp.12-19
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    • 2004
  • This study was performed to investigate the optimum condition of protease production from submerged culture of oak mushroom (Lentinula edodes, Sanlim No. 5) and its enzymatic features. Among several combinations of media, the combination of wheat bran, corn flour, water and corn oil (WB+CF+W+ CO) yielded 84.8 U/g of maximum protease activity. This combination of ingredients, in spite of not being particularly protein-rich in comparison to the other media, allowed for good growth of the fungus and maximal protease production. Comparison of different growth medium liquids indicated that demineralized water afforded the best growth of the fungus and the highest protease activity. Acetate buffer and acidified water negatively affected The protease production peaked around 72 hr of incubation, and decreased thereafter. The molecular weights of produced protease were about 45,000 by Sephadex G-75 chromatography. The pH optimum for protease activity was 4, while maximal activity incubated at 37℃ for 1 hr was observed between pH 4~6. The optimum temperature of this protease was 55℃, and the enzyme was active over a broad temperature range (30~60℃), indicating that this protease would be suitable for a wide range of applications where. different pH and temperature are necessary, such as digestive aids, food industry, beer and tannery industries.

Protease released during germination of dictyostelium discoideum spores (Dictyostelium discoideum의 포자가 발아중에 유출하는 protease에 관하여)

  • 윤경하;윤철호
    • Korean Journal of Microbiology
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    • v.27 no.1
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    • pp.43-47
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    • 1989
  • Characteristics and roles of protease released during the germination of Dictyostelium discoideum spores were investigated. When geat activated, the spores germinated, progressively releasing the protease into the extracellular medium. The protease activity exhibited high at pH 2.5. When cyclogeximide was added to culture, complete germination (emergence) and protease release were stopped. Addition of purified nonspecific protease to culture speeded up germination. These results suggest that excreted protease may play a role in removal of the spore wall.

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The Synergistic Action of the AL-Protease from Arthrobacter luteus on the Lysis of Yeast Cell Walls (Arthrobacter luteus가 생산하는 AL-Protease의 효모세포벽 용해 촉진작용)

  • Oh, Hong-Rock;Funatsu, Masaru
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.14 no.4
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    • pp.401-408
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    • 1985
  • The yeast cell wall lytic action of the alkaline AL-protease, which was found out of the crude Zymolyase that a kind of yeast cell wall lytic $endo-{\beta}-1$, 3-glucanase produced from Arthrobacter luteus, was investigated with the viable cells of S. sake and it's cell wall preparation. AL-protease on the lysis of the viable yeast cells showed very low activities with the alone, but the lytic activities were highly increased with the combination of AL-protease and Zymolyase. On the stepwise treatment of the viable yeast cells with AL-protease and Zymolyase, the cells were lysed highly only by the course having a treatment with Zymolyase after pretreatment with AL-protease. Thus synergistic action of AL-protease was not observed with any some commercial enzymes, known as a type of alkaline and serine protease such as AL-protease, and was also found to be affected greatly by the culture conditions and species of the yeast tested. AL-protease caused the release of some peptide and a lot of sugar from the cell wall preparation, but could not lysed the cell wall more than 66%. Whereas Zymolyase could lysed the cell walls almost completely with alone. On the basis of these results, the synergistic action of AL-protease on the lysis of S. sake cells is hypothesized that at first AL-protease bind to the yeast cell surface layer consisting of mannan and protein, and then changes their conformation to facilitate the penetration of Zymolyase from the outside to the inside framework layer constituted of alkali insoluble ${\beta}-1,\;3-glucan$.

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Protease Inhibitor Production using Streptomyces sp. SMF13

  • Kim, In-Seop;Kim, Hyoung-Tae;Lee, Hyun-Sook;Lee, Kye-Joon
    • Journal of Microbiology and Biotechnology
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    • v.1 no.4
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    • pp.288-292
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    • 1991
  • The aim of the current study is to evaluate the effects of medium compositions on the production of protease inhibitor in Streptomyces sp. SMF13. The production of protease inhibitor was counter-currently linked to extra-cellular protease, which were regulated by the culture conditions. Nitrogen source was the most critical ingredient affecting the production of protease inhibitor and protease. Carbon source was an important factor to determine the culture pH which affected very clearly the formation of protease and protease inhibitor. Inorganic phosphate inhibited the protease inhibitor production which was linked to the cell growth rate, although the optimal conditions for the production of protease inhibitor were not favouring to the cell growth.

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Characteristics of Soybean Hydrolysates Prepared with Various Protease (단백질 분해효소 종류에 따른 콩 가수분해물의 특성)

  • Jeong, Kyu-Ho Jeong;Seo, Ji-Hyung;Jeong, Yong-Jin
    • Food Science and Preservation
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    • v.12 no.5
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    • pp.460-464
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    • 2005
  • In order to produce functional soy hydrolysates, we investigated the characteristics of soy hydrolysates prepared with 4 kinds of commercial proteases. The yield was high in protease(B), in which 43.2% soy flour and 61.6% SPI were obtained. The solubility and the contents of total phenolic compound were greatly increased by the treatment of protease(B) along with protease(C). The calcium intolerance was improved after the protease(B) treatment in soy flour or Soybean Protein isolate (SPI). Consideration for the physicochemical characteristics including yield, protease(B) has potential application for the production of soy hydrolysates. After the protease treatment, the beany flavor of soy flour became weak and the bitter taste was strong in both soy flour and SPI. However, there was no difference of beany flavor and bitter taste among delete protease hydrolysates. Nevertheless, further modifications and improvements to the sensory characteristics would be required for the development of a range of products with the hydrolysate.

Cloning of a Alkaline Protease Gene from Xanthomonas sp. YL-37 (Xanthomonas sp. YL-37의 Alkaline Protease 유전자의 클로닝)

  • 이대희;김수경;이승철;윤병대;황용일
    • Microbiology and Biotechnology Letters
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    • v.23 no.2
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    • pp.145-149
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    • 1995
  • For the purpose of developing a new biodegradable detergent, we have isolated a gene encoding wide-range temperature applicable alkaline protease from Xanthomonas sp. YL-37 (Lee et al., 1994, Kor. J. Appl. Microbiol. Biotechnol.). An alkaline protease gene was isolated from the gene bank that was prepared from the chromosomal DNA of Xanthomonas sp. YL-37. From the results of agarose gel electrophoresis and a restriction enzyme mapping, a 2.7 kb DNA fragment containing the alkaline protease gene was inserted in the plasmid pUC9. Extracellular activity of a clone having alkaline protease gene was detected on SDS-polyacrylamide gel with activity staining assay. The molecular weight of alkaline protease was determined to be about 64 kDa from 11% SDS-PAGE analysis. Alkaline protease activity, produced from E. coli which harboring the plasmid, showed no difference at reaction temperature 20, 30 and 40$\circ$C, respectively. This result showed that alkaline protease produced from E. coli harboring the plasmid was apparently the same as that of Xanthomonas sp. YL-37.

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On a highly proteolytic mutant strain of Aspergillus flavus (Aspergillus flavus의 강력 protease생성 돌연변이의 유발)

  • 이영녹;박용근;고상균
    • Korean Journal of Microbiology
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    • v.18 no.2
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    • pp.51-58
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    • 1980
  • Mutational experiments were performed to improved to improve the protease productivity of Aspergillus flavus KU 153, which is selected among the wild strains. A UV-induced mutant strain having high protease productivity was obtained by the use of the clear zone method as a simple criterion for a primary screening test. Neutral and alkaline protease activities of hte mutant strain were higher than 1.8 times, comopared with those of the parental strain, respectively, while in the case of acid protease, it was 2.7 times. The mutant strain selected was more powerful in the production of cellulase and amylase, as well s protease in wheat bran, compared with those of the parental strain. protease production of the parental strain has reached maximum level at 3 days culture, while alkaline nad neutral protease production of the mutantstrain has reached at 2 days culture. On the other hand, the mutant strain formed the spore slowly, compared with the parental strain. Column chromatography of the neutral protease on DEAE-Sephadex A-50 showed that the mutant strain was not induced the formation of another neutral protease isozyme, but induced the variation in the function of regulatory gene.

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Protease의 수준별 급여가 육계의 생산성, 혈액의 성상, 장기 무게 및 단백질 소화율에 미치는 영향

  • 박재흥;류명선;나종삼;김종승;류경선
    • Proceedings of the Korea Society of Poultry Science Conference
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    • 2002.11a
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    • pp.122-123
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    • 2002
  • An experiment was conducted to investigate the effect of dietary supplemental protease on performance, blood components, organ weight and protein digestibility. Two hundred and fifty, one day old cobb x cobb male broiler chicks were assigned to 4 treatments with 4 replicates. Four levels of protease were supplemented with 0, 40, 80 and 160U. Weight gain, feed intake, feed conversion were weekly measured for 5 weeks. Blood components and organ weight were examined at the end of experiment. Metabolic feeding trial was carried out to investigate the protein digestibility for one week at five weeks of age. Basal diets contained 21.5, 19.0% CP and 3,100, 3,200kca1/kg ME for starter and finisher, respectively. Weight gain of chicks fed 80U protease was significantly higher than other treatments for starting period(P〈0.05). Feed conversion of chicks fed protease addition diets was improved as dietary protease increased. It showed significant difference between 80U and control groups(P〈0.05). Sera protein tended to be lowered in protease added groups. HDL-cholesterol was decreased at the maximum level of protease. However, organ weight and protein digestibility were not influenced significantly by dietary protease level. In conclusion, the present study demonstrated that protease addition in broiler diets improved the feed conversion.

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Purification and Characteristics of Protease Produced by Syncephalastrum racemosum PDA 132-2 from Korean Traditional Meju (메주 유래의 Syncephalastrum racemosum PDA 132-2가 생산하는 Protease의 정제 및 특성)

  • 유진영;임성일
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.28 no.5
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    • pp.1010-1016
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    • 1999
  • Protease related mold was isolated and selected as a starter culture for commercial production of meju. Isolated microorganism was identified as Syncephalastrum racemosum PDA 132 2. To obtain basic data about protease for production of soybean peptides and application of the strain in meju fermentation, we extrated and purified protease and charateristics of the enzyme were investigated. The optimum condition for the production of enzyme was pH 4.0, 30oC, 5 days. The protease was purified 19.7 folds by gel filtration and ion exchange chromatography and specific activity was 12.4unit/mg. The purified enzyme was 34kDa in size, thiol protease(100% inhibited by PCMB), and was acidic protease(stable between pH 2.0~5.0). Vmax of the enzyme was 2.14 g/min which was lower(1/50) than that of by Asp. wentti and B. subtilis.

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