• Title/Summary/Keyword: protease activities

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Influence of the Kilning Conditions on Enzymatic Activity of Rice (Oryza sativa) Malt

  • Nguyen, Thach Minh;Nguyen, Xich Lien;Hoang, Kim Anh;Lee, Soo
    • Journal of the Korean Applied Science and Technology
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    • v.26 no.1
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    • pp.10-17
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    • 2009
  • This study investigated the effect of kilning condfition on the diastatic power and activities of protease, $\alpha$-amylase, and $\beta$-amylase in rice malt. Common rice (Oryza sativa) was steeped at $30^{\circ}C$ for 50 h, germinated at $30^{\circ}C$ for 7 days, and kilned at $50^{\circ}C$ for 24 h. The moisture content and enzymatic activities were determined under various kilning times. As a result, the moisture content was reduced from 42.1 % to 3.9% after 24 h of kilning at $50^{\circ}C$. The protease activity of rice malt showed lower value than that of barley malt. All enzymatic activities were decreased during the kilning stage. Results indicated that after prolonged kilning at $50^{\circ}C$, the inactivation of hydrolytic enzymes might be occurred. Even though the amylolytic activity of malted rice showed low value, the rice malt shows the potential characteristics as ingredient for the brewing and cereal industries.

Evaluation of Extracellular Enzyme Activity of Fungi from Freshwater Environment in South Korea (담수환경에서 분리한 곰팡이의 세포외분해효소 활성 탐색)

  • Hye Yeon Mun;Yoosun Oh;Jaeduk Goh
    • The Korean Journal of Mycology
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    • v.51 no.4
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    • pp.265-276
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    • 2023
  • This study aimed to isolate and characterize fungi from freshwater environments in South Korea and evaluate their extracellular enzyme activities. Fungal strains were collected from various freshwater sources and identified using phylogenetic analysis. The isolated fungi included known aquatic hyphomycetes and previously unreported species. Extracellular enzyme, including those of protease, amylase, lipase, cellulase, laccase, and chitinase, activities were evaluated. Among the isolated strains, several showed high enzyme activity, suggesting their potential role in organic matter decomposition in freshwater ecosystems. This research expands our knowledge of the diversity and enzyme activities of the fungi in freshwater environments, contributing to our understanding of their ecological roles.

Characterization of Alkaline Serine Proteases Secreted from the Coryneform Bacterium TU-19

  • Kang, Sun-Chul;Park, Sang-Gyu;Choi, Myong-Chul
    • Journal of Microbiology and Biotechnology
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    • v.8 no.6
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    • pp.639-644
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    • 1998
  • Extracellular serine proteases were isolated from a soil bacterium, alkalophilic coryneform bacterium TU-19, which have been grown in a liquid medium optimized at 3$0^{\circ}C$ and pH 10.0. Three different sizes, 120 kDa (protease I), 80 kDa (protease II), and 45 kDa (protease III), of serine pro teases were purified using Sephadex G-150 and QAE-Sephadex chromatography (Kang et al. 1995. Agric. Chem Biotech. 38: 534-540). SDS-PAGE showed that the 120 kDa protease was degraded into the 80 kDa protease in 20 mM Tris-HCI (pH 8.0) buffer solution. This degradation was enhanced in the presence of 0.5 M NaCl and 5 mM EDTA, but was inhibited in the presence of 5 mM $CaCl_2$. These results indicated that the $Ca^{2+}$ ion seems to stabilize the 120 kDa protease like other proteases derived from Bacillus species. The $NH_2$-terminal amino acid sequences of the 10 residues of both proteases were completely identical: Met-Asn-Thr-Gln-Asn-Ser-Phe-Leu-Ile-Lys. In contrast to this, the 80 kDa protease has 1.5 times higher specific activity than the 120 kDa protease does (Kang et al. 1995. Agric. Chern. Biotech. 38: 534-540). Therefore the C-terminal of the 120 kDa protease seems to be autolyzed to the 80 kDa protease but this autolysis did not decrease the protease activity. Optimum pH and temperature of both 80 kDa and 120 kDa proteases were pH 10.5 and $45^{\circ}C$, respectively, and pH and thermal stability were almost identical. Several divalent ions except the $Fe^{2+}$ ion showed similar effects on activities of both proteases, which are similarly resistant to three different detergents.

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Proteases and Antioxidant Activities of Doenjang, Prepared with Different Types of Salts, during Fermentation (소금 종류를 달리하여 제조한 된장들의 발효 중 protease 역가 및 항산화 활성 변화)

  • Shim, Jae Min;Lee, Kang Wook;Kim, Hyun-Jin;Kim, Jeong Hwan
    • Microbiology and Biotechnology Letters
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    • v.44 no.3
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    • pp.303-310
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    • 2016
  • In this study, doenjang samples were prepared with different types of salts (12%, w/w): purified salt (PS), 3-year aged solar salt (SS3), 1-year aged solar salt (SS1), and bamboo salt melted 3 times (BS). Whole-soybean mejus were fermented with starters consisting of 2 Bacillus strains, a yeast, and a fungus (starter doenjang), and control mejus were fermented with organisms present naturally in rice straw (non-starter doenjang). The whole-soybean mejus were dried, and then mixed with cooked soybeans and the respective salts. The doenjang samples were fermented for 13 weeks at 25℃. The protease (acid, neutral, and alkaline) activities, fibrinolytic activities, and antioxidant capacities of the samples were examined every week. BS doenjang showed the highest acid protease (6.46 ± 0.20 unit/g) and fibrinolytic activities (0.61 unit/ml). Among the starter doenjang samples, those made with SS and BS showed the highest total phenolic contents after 91 days of fermentation. For antioxidant activities, SS3 doenjang showed higher activities than the other doenjang samples, as evaluated by ABTS, DPPH, and FRAP assays. These results suggest that solar salt, especially aged for 3 years, is better than purified salt in terms of producing better functionalities of doenjang.

Effects of the ratio of raw materials on the quality and taste of soy sauce - (1) Studies on the changes of Enzymatic activities and chemical components during Koji Preparation with various ratio of raw materials - (원료배합(原料配合)이 간장의 품질(品質)과 풍미(風味)에 미치는 영향(影響) - 제1보(第一報) 원료배합(原料配合)을 달리한 고일(一)지제조중(製造中)의 효소역가(酵素力價)및 성분변화(成分變化)에 관(關)한 실험(實驗) -)

  • Kim, Yong-Hwi;Kim, Jae-Uk
    • Applied Biological Chemistry
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    • v.4
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    • pp.17-22
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    • 1963
  • Changes of Enzymatic activities and chemical components during Koji preparation of soy sauce with various ratio of soy bean and wheat were studied as first step for checking the current ratio of raw materials for improved soy sauce and determination of proper ratio of them in the respect to its quality and taste, and following results could be obtained. 1. The Protease in the dryed Koji were mainly conmposed of a part which active at the neutral (about pH 6.0) range, while parts which active at acid and alkaline side were inferior. The more amount of wheat increases as raw materials of Koji, the stronger Protease activities of acid and neural side were, while the weaker alkaline side were. 2. Activity of Enzymes were increased rapidly in earlier stage then gradually in later stage or Protease and ${\beta}-Amylase$ rapidly throughout except drying of ${\beta}-Amylase$ during the course of Koji preparation. The more amount of wheat as raw material increases, the stronger Protease and ${\beta}-Amylase$ activity except. ${\beta}-Amylase$ were. 3. Reducing sugar, amino nitrogen, total nitrogen were increased, while total sugar were decreased during the course of Koji preparation. 4. The more amount of wheat increases as raw materials, the more increase reducing sugar, total sugar were, while the total nitrogen were decreased, no noticeable differences were observed in the amino nitrogen among the dryed Kojies.

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Changes in Microflora and Enzymes Activities of Traditional Kochujang Prepared with Various Raw Materials (담금원료에 따른 전통식 고추장의 숙성 중 미생물과 효소력의 변화)

  • Shin, Dong-Hwa;Kim, Dong-Han;Choi, Ung;Lim, Mi-Sun;An, Eun-Young
    • Korean Journal of Food Science and Technology
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    • v.29 no.5
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    • pp.901-906
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    • 1997
  • In order to reproduce and improve quality of traditional kochujang, various raw materials were added to prepare kochujang by replacing part of the glutinous rice. Chemical composition, microbial characteristics and enzyme activities were investigated during fermentation. Crude protein and salt contents of kochujang did not change significantly during fermentation, but moisture contents increased linearly. The pH and titratable acidity of kochujang changed little in garlic added group. The viable cell counts of aerobic bacteria and yeasts in the kochujang increased until 60 days of fermentation and then decreased slowly except for the garlic added group in which they increased during the last period of fermentation. Aerobic bacterial count did not show any remarkable differences among the samples and slowly decreased after 60 days of fermentation. The activities of liquefying and saccharifying amylases decreased until 45 days, but increased at 60th day. Acidic protease activities of each group were strong during the initial period, but neutral protease showed the highest activity from the 30 to 45 days of fermentation. Protease activities increased by addition of soy sauce, Chinese matrimony vine and purple sweet potato.

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Effect of Temperature on Growth Rate and Protease Activity of Antarctic Microorganisms (극지 미생물들의 배양온도에 따른 성장률 및 protease activity 영향 연구)

  • Kim, Hyun-Do;Choi, Jong-Il
    • Microbiology and Biotechnology Letters
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    • v.42 no.3
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    • pp.293-296
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    • 2014
  • This study was conducted to investigate the effect of culture temperature on the growth rate and protease activity of Antarctic microorganisms. The Antarctic microorganisms PAMC 25641, 25614, 25719 and 25617 were obtained from the Polar and Alpine Microbial Collection (PAMC) at the Korea Polar Research Institute. These microorganisms were confirmed for the excretion of protease on a plate with skim milk. The identification of microorganisms was carried out using the 16S rDNA sequencing method. PAMC 25641 showed the highest protease activity among the subjects tested, and PAMC 25617 exhibited the highest growth rate. The growth rates of the microorganisms were not affected by temperature, except for PAMC 25617. However, protease activities were increased for all strains in a temperature dependent fashion. These results suggest the possible application of Antarctic microorganisms for the efficient production of low temperature proteases.

Purification and Characterization of a Novel Serine Protease with Fibrinolytic Activity from Tenodera sinensis (Chinese Mantis) Egg Cases

  • Cho, So-Yean;Hahn, Bum-Soo;Kim, Yeong-Shik
    • BMB Reports
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    • v.32 no.6
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    • pp.579-584
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    • 1999
  • Mantis egg fibrolase (MEF-3) was purified from the egg cases of Tenodera sinensis using ammonium sulfate fractionation, gel filtration on Bio-Gel P-60, DEAE Affi-Gel blue gel affinity chromatogragphy, and MONO-Q anion-exchange chromatography. This protease had a molecular weight of 35,600 Da as determined by SDS-polyacrylamide gel electrophoresis under reducing conditions and its isoelectric point was 6.0. The N-terminal amino acids sequence was Ala-Thr-Gln-Asp-Asp-Ala-Pro-Pro-Gly-Leu-Ala-Arg-Arg. This sequence was 80% homologous to the serine protease from Tritirachium album. MEF-3 readily digested the ${\alpha}$-and ${\beta}$-chains of fibrinogen and more slowly the ${\gamma}$-chains. It showed strong proteolytic and fibrinolytic activities. Phenylmethanesulfonyl fluoride and chymostatin inhibited its proteolytic activity, while EDTA, EGTA, cysteine, ${\beta}$-mercaptoethanol, elastinal, tosyl-lysine chloromethylketone, and tosyl-amido-2-phenylethyl chloromethyl ketone did not affect its proteolytic activity. Among the chromogenic protease substrates, the most sensitive one to the hydrolysis of MEF-3 was benzoyl-Phe-Val-Arg-p-nitroanilide. Based on these experimental results, we speculated that MEF-3 is a serine protease with a strong fibrin(ogen)olytic activity.

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Characterization of Symbiotic Bacteria from Entomopathogenic Nematode (곤충병원성 선충로부터 분리된 공생박테리아의 종별 특성)

  • 박선호;김지연
    • KSBB Journal
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    • v.17 no.3
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    • pp.276-282
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    • 2002
  • Symbiotic bacteria with highly effective insecticidal activities were isolated and compared with their physiological characteristics from seven species of entomopathogenic nematodes belong to Steinernamatidae and Heterorhabditidae sp., and three of them were identified as Xenorhabdus nematophilus. Culture characteristics, insecticidal activities, pretense activities and fatty acid contents of various symbiotic bactierial isolates were also examined. In the case of cell growth and insecticidal activity, XR-PC and XR-MK were superior to other species when cultured in vitro. The insecticidal activity were highest at the early exponential growth phase, and gradually decreased with time. The protease activity of XR-DR was remarkable compared to other species. In the case of HE-HY, however the pretense activity increased in parallel with cell growth. Interestingly, the fatty acid patterns of Xenorhabdus nematophilus isolated from different emtomopathogenic nematode, showed remarkable differences in their contents of 12:0, 14:0, 16:1 cia 5 and 17:0 cyclo and hydroxy and branch factty acids were varied from 2% to 15% among total fatty acid contents.

Isolation and Enzymatic Characterization of Bacteria from Livestock Manure (축산분뇨에서 분리한 세균의 동정 및 효소학적 특성)

  • 김진선;정소선;이준석;최미영;서승염
    • Korean Journal of Microbiology
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    • v.37 no.3
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    • pp.214-220
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    • 2001
  • To develop the effective composting system, we isolated bacteria that have the abilities to degrade organic matters such as cellulose, carbohydrate, protein and lipid during the compositing of livestock manureAmong 24 strains, 6 bacteria have all the enzymatic activities of protease, amylase, cellulase and lipase.These microorganisms were identified as Corynebacterium varibilis, Bacillus spp., Pseudomonas spinosa,Acetobacter calcoaceticus and Athrobacter cumminsii All the enzymes produced by the bacteria showedactivities at the broad pH range and the maximal activities were obtained at $60^{\circ}C$. It seemed that after theincrease of temperature caused by fermentation of livestock manure, the enzymes started to degrade the rawmaterials, which are added for the control of humidity. However cellulase activity was maximum at $37^{\circ}C$,suggesting that the cellulase-producing bacteria work at an early stage of livestock manure fermentation toprovide the organic material for the growth of other bacteria. The production of the enzymes were growth-associated and maximal activities appeared at the early stationary phase of growth.

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