• Title/Summary/Keyword: enzymatic characteristics

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Characteristics of ${\beta}-galactosidase$ activity in Lactobacillus plantarum from kimchi (김치에서 분리한 유산균 Lactobacillus plantarum의 ${\beta}-galactosidase$ 특성에 관한 연구)

  • Kang, Me-Seon;Rhee, Young-Hwan
    • Applied Biological Chemistry
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    • v.39 no.1
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    • pp.60-66
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    • 1996
  • To investigate a possible application of three strains of lactic acid bacteria(strain No. 49. No. 61. No. 75) from kimchi in milk fermentation industry, the optimal condition for production of intracellular ${\beta}-galactosidase$ from Lactobacillus(L.) plantarum and its enzymatic properties were examined. The preferable carbon source of the medium for strain No. 49 in production of ${\beta}-galactosidase$ was MRS broth with 1.0% lactose instead of dextrose of pH 65. for strain No. 75 with 1.0% galactose and for strain No. 61 with 3.0% lactose at pH 7.5, respectively. The maximum enzyme production from strain No. 49, No. 75 was observed after 48 hours culture at $30^{\circ}C$ in a medium containing the appropriate carbon source, from strain No. 61 after 48 hours culture at room temperature. The optimum temperature for ${\beta}-galactosidase$ activity from L. plantarum was $60^{\circ}C$ for strain No. 49, $37^{\circ}C$ for strain No. 61 and $50^{\circ}C$ for strain No. 75, respectively. The heat stability of enzyme activities for all three strains remained 90% at $45^{\circ}C$. The optimal pH was pH 6.5 and enzyme activities were most stable at pH for all three bacteria.

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Overexpression and Characterization of Bovine Pancreatic Deoxyribonuclease I in Saccharomyces cerevisiae and Pichia pastoris (Saccharomyces cerevisiae와 Pichia pastoris에서 Bovine Pancreatic Deoxyribonuclease I의 과발현과 특성)

  • Cho, Eun-Soo;Kim, Jeong-Hwan;Yoon, Ki-Hong;Kim, Yeon-Hee;Nam, Soo-Wan
    • Microbiology and Biotechnology Letters
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    • v.40 no.4
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    • pp.348-355
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    • 2012
  • In the present study, we investigated the overexpression and characterization of bovine pancreatic (bp)- DNase I in Saccharomyces cerevisiae and Pichia pastoris. The bp-DNase I gene was fused in frame with the GAL10 promoter, $MF{\alpha}$, and GAL7 terminator sequences, resulting in the plasmid, pGAL-$MF{\alpha}$-DNaseI (6.4 kb). Also, the bp-DNase I gene was fused in frame with the AOX1 promoter, $MF{\alpha}$, and AOX1 terminator sequences, resulting in the plasmid, pPEXI (8.8 kb). The recombinant plasmids, pGAL-$MF{\alpha}$-DNaseI and pPEXI were introduced into S. cerevisiae and P. pastoris host cells, respectively. When the transformed yeast cells were cultured at $30^{\circ}C$ for 48 h in galactose or methanol medium, bp-DNase I was overexpressed and the most of activity was found in the extracellular fraction. P. pastoris transformant activity showed 45.5 unit/mL in the culture medium at 48 h cultivation, whereas S. cerevisiae transformant revealed 37.7 unit/mL in the extracellular fraction at 48 h cultivation. The enzymatic characteristics, such as DNA cleavage and half life were investigated. Treatment of the recombinant DNase I from P. pastoris induced degradation of the calf thymus DNA within 1 minute, and this DNA degradation rate was higher than that of commercial bp-DNase I (SIGMA) and the recombinant DNase I from S. cerevisiae.

Enzymatic Characteristics for Xylanase Activity of Tremella fuciformis and its Symbiotic Fungi (흰목이버섯 및 공생균이 분비하는 Xylanase 효소적 특성)

  • Chang, Hyun-You;Kim, Gwang-Po;Hong, In-Pyo;Kim, Han-Kyoung;Chung, Jong-Cheon;Sung, Jae-Mo
    • The Korean Journal of Mycology
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    • v.25 no.4 s.83
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    • pp.297-304
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    • 1997
  • Effects of cultural conditions on the production of xylanase by Tremella fuciformis, symbiotic fungi and mixed fungi were investigated. The optimum carbon source for high production of xylanase by T. fuciformis, symbiotic fungi and mixed fungi was xylose. The optimum nitrogen source for both T. fuciformis and symbiotic fungi was $KNO_3$, whereas mixed fungi was $(NH_4)_2SO_4$. The optimum culture period for high production of xylanase was 5 days for both T. fuciformis and mixed fungi, and 6 days for symbiotic fungi, respectively. The optimum temperature for T. fuciformis and symbiotic fungi was $40^{\circ}C$, and the corresponding value for mixed fungi was $45^{\circ}C$. Xylanase activity was high at pH 6 for T. fuciformis and symbiotic fungi, and pH 7 for mixed fungi. Except $Hg^{2+}$ and $Pb^{2+}$, metal ions in T. fuciformis inhibited the activity of xylanase, and, thermal stability of xylanase in T. fuciformis, symbiotic fungi and mixed fungi maintained 80% of activity until $50^{\circ}C$. The Michaelis constant (Km) of xylan was $6.25{\times}10^{-5}\;M$ in T. fuciformis, $5.6{\times}10^{-2}\;M$ in symbiotic fungi, $5.2{\times}10^{-2}\;M$ in mixed fungi.

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Characteristics of the Absorption of Cherry Tomato by the Application of Chelated Calcium and Germanium (킬레이트화 칼슘 및 게르마늄의 방울토마토 시용에 따른 흡수 특성)

  • Jang, Young-Hee;Lee, Seung-Hwan;Park, Young-Il;Lee, Kyu-Seung
    • Korean Journal of Soil Science and Fertilizer
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    • v.45 no.5
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    • pp.787-791
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    • 2012
  • Bloods from slaughtered Korean native cattle (Hanwoo : Bos taurus coreanae) can be used in agriculture with various beneficial effects on the environment. This study was performed to find out the effect of the application of calcium and germanium, which are chelated with peptides made of cattle bloods on the absorption of them by cherry tomato (Lycopersicon esculentum Mill.). Peptide was purified by enzymatic hydrolysis and ultrafiltration from serum protein, which is composed of Asp, Glu, Lue etc. Chelated Ca and Ge were produced by reacting the peptide with $CaCl_2$ and Ge-$132^{(R)}$. Then, they were applied to cherry tomato during cultivation period at the rate of $300mL{\times}3times$ (10 days interval) for each tomato tree. Application of chelated-Ca increased the Ca contents in leaves and fruits of cherry tomato showing as 19.9% and 23.4% in newerly prepared chelated-Ca-200 ($200mg\;L^{-1}$), 8.1% and 6.8% in commecrial Calciolid Ca-300 ($300mg\;L^{-1}$) compared to 3152.6 and $63.2mg\;kg^{-1}$ in control, respectively. Application of chelated-Ge showed the increase in the germanium contents in both leaves and fruits of cherry tomato by over 6 times than those of control and over 4 times than Ge $132^{(R)}$-20 application. As a result of this study, it seemed that the application of chelated-Ca and chelated-Ge would be more effective than the existing commertial one in the absorption of calcium and germanium by cherry tomato.

Quality Characteristics of Garlic Suspensions Using Protopectin Hydrolytic Enzymes (Protopectinase를 이용하여 제조한 마늘 단세포화물의 품질특성)

  • Baek Ku-Hyoun;Kim Sung-Soo;Tak Sang-Bum;Kang Byung-Sun;Kim Dong-Ho;Lee Young-Chun
    • Food Science and Preservation
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    • v.13 no.3
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    • pp.351-356
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    • 2006
  • This study was carried out to investigate the change of functional component and volatile flavor components from garlic for which both were treated with protopectinase (PPase) and mechanical maceration during storage period. Alliin content of gallic suspensions macerated mechanically were 11.0 mg/g at 0 day and 6.6 mg/g at 24 day. Whereas alliin content of garlic treated with PPase were 8.5 m/g at 0 day and 7.0 mg/g at 24 day. Importantly, over 40% of alliin which is the most unstable component during the mechanical maceration remained with an intact form for 24 day after the enzymatic treatment. The flavor component from gallic suspensions were extracted by solid-phase microextraction (SPME) and were analyzed and identified by gas chromatography (GC) and chromatography/mass spectrometry (GC/MS). The number and concentrations of flavor components of gallic macerated mechanically were increased during storage period, and total 18 kinds of flavor compounds were identified. Thus, the PPase treatment of garlic could be a better choice for preparation of the highly valuable and functional processed food as well as for prolonging the preservation period.

Selection of Kimchi Starters Based on the Microbial Composition of Kimchi and Their Effects (김치 미생물 조성을 바탕으로 한 김치 스타터의 선정 및 효과)

  • Jin, Hyo-Sang;Kim, Jong-Bum;Yun, Yeong-Ju;Lee, Kyung-Ja
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.37 no.5
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    • pp.671-675
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    • 2008
  • Based on information about the major microbial composition of kimchi and its relation to the taste, Leuconostoc mesenteroides K2M5 and Lactobacillus sakei K5M3 were selected as kimchi starter candidates. These two strains were found to be safe for industrial use because they showed neither harmful characteristics like ${\beta}$-hemolysis, ammonia and indole formation, and gelatin liquefaction, nor enzymatic activities like phenylalanine deaminase, ${\beta}$-glucuronidase, ${\beta}$-glucosidase, 7${\alpha}$-dehydroxylase and nitroreductase. Starter kimchi made with these strains were better in taste than the conventional kimchi when they are evaluated both by laboratory personnel and the public. Also microbial analysis of starter kimchi showed only starter bacteria after they were fermented to have the optimum acidity. Starter kimchi prepared with these two strains were not much different in physicochemical properties to the conventional kimchi except in that the starter kimchi were much higher in volatile organic acid content such as lactic acid. These results suggest that kimchi quality can be controlled to have consistent properties, both in taste and microbial composition, by using bacterial starters.

Characterization of TNP-cellulose as Substrate for Cellulase Assay (TNP-cellulose의 섬유소 분해효소 활성도 측정을 위한 기질로서의 특성)

  • Maeng, Jeong-Seob;Nam, Yoon-Kyu;Choi, Woo-Young
    • Korean Journal of Agricultural Science
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    • v.21 no.2
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    • pp.142-147
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    • 1994
  • Characteristics of TNP-cellulose which prepared from carboxymethyl cellulose powder, CM32, as substrate for cellulase activity assay were investigated. Enzymatic hydrolysis of TNP-cellulose occured on the cellulose moiety but not on amide bonds, following Michaelis-Menten kinetics. Three cellulase preparations from Trichoderma viride, Aspergillus niger, and Cellulomonas sp. were tested for their pH and temperature dependences and compared with the method determining the increase in reducing power. The enzyme activity was found to have the same temperature range in both methods, however the pH range was broadened in the case of using TNP-cellulose as substrate. The colorimetric method for cellulase assay using TNP-cellulose as substrate was compared with the other methods: one based on determination of the increase in reducing power; and the other based on determining the decrease in viscosity of Na-CM-cellulose solution. The activities measured by the colorimetric method showed a linear correlation with the enzyme concentration of certain range in all three enzymes tested, and the activity values were proportional to those obtained from the other methods. Depending on the enzyme, however, the activity values from this method were not always in proportion to those from the viscometric method. suggesting that this method was not specific for determination of the endo-type cellulase.

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Comparison of Enzymatic Activity and Cleavage Characteristics of Trypsin Immobilized by Covalent Conjugation and Affinity Interaction (공유결합과 친화력결합에 의한 고정화 Trypsin의 효소역가와 절단특성 비교)

  • Jang, Dae-Ho;Seong, Gi-Hun;Lee, Eun-Kyu
    • KSBB Journal
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    • v.21 no.4
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    • pp.279-285
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    • 2006
  • We investigated the effects of immobilization chemistry on the yield of immobilization and the bioactivity of the immobilized enzymes. Trypsin as a model protein and macroporous polymer beads(Toyopearl AF 650M, Tosho Co., Japan) was used as a model matrix. Four methods were used to immobilize trypsin; covalent conjugation by reductive amination(at pH 10.0 and pH 4.0) and affinity interaction via streptavidin-biotin, and double-affinity interaction via biotin-streptavidin-biotin system. The covalent conjugation immobilized $3{\sim}4$ mg/ml-gel, ca. 3-fold higher than the affinity method. However, the specific activity of the covalently(pH 10.0) and affinity-immobilized trypsin(via streptavidin-biotin) are ca. 37% and 50%, respectively, of that of the soluble enzyme(on the low-molecular-weight BAPNA substrate). When the molecular size of a substrate increased, the affinity-immobilized trypsin showed higher clavage activity on insulin and BSA. This result seemed to indicate the streptavidin-biotin system allowed more steric flexibility of the immobilized trypsin in its interaction with a substrate molecule. To confirm this, we studied the molecular flexibility of immobilized trypsin using quartz crystal microbalance-dissipation. Self-assembled monolayers were formed on the Q-sensor surface by aminoalkanethiols, and gultaraldehyde was attached to the SAMs. Trypsin was immobilized in two ways: reductive amination(at pH 10.0) and the streptavidin-biotin system. The dissipation shift of the affinity-immobilized trypsin was $0.8{\times}10^{-6}$, whereas that of the covalently attached enzyme was almost zero. This result confirmed that the streptavidin-biotin system allowed higher molecular flexibility. These results suggested that the bioactivity of the immobilized enzyme be strongly dependent on its molecular flexibility.

Factors Influencing the Acrylamide Content of Fried Potato Products

  • Jin, Yong-Ik;Park, Kyeong-Hun;Chang, Dong-Chil;Cho, Ji-Hong;Cho, Kwang-Su;Im, Ju-Sung;Hong, Su-Young;Kim, Su-Jeong;Nam, Jung-Hwan;Sohn, Hwang-Bae;Yu, Hong-Seob;Chung, Ill-Min
    • Korean Journal of Environmental Agriculture
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    • v.35 no.4
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    • pp.247-255
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    • 2016
  • BACKGROUND: Acrylamide (CAS No. 79-06-1) is known to be a carcinogenic compound, and is classified as a Group 2A compound by the International Agency for Research on Cancer (IARC, 1994). Acrylamide can be generated during the browning process via the non-enzymatic Maillard reaction of carbohydrates such as reducing sugars and of amino acids such as asparagine, both of which occur at a temperature above $120^{\circ}C$. Potato tubers contain reducing sugars, and thus, this will affect the safety of processed potato products such as potato chips and French fries. In order to reduce the level of acrylamide in potato processed products, it is therefore necessary to understand factors that affect the reducing sugar content of potatoes, such as environmental factors and potato storage conditions, as well as understanding factors affecting acrylamide formation during potato processing itself. METHODS AND RESULTS: Potatoes were cultivated in eight regions of Korea; For each of these different environments, soil physico-chemical characteristics such as pH, electrical conductivity, total nitrogen, available phosphate, and exchangeable cation content were measured and correlations with potato reducing sugar content and potato chip acrylamide levels were examined. The reducing sugar content in potato during storage for three months was determined and acrylamide level in potato chip was analyzed after processing. The storage temperature levels were $4^{\circ}C$, $8^{\circ}C$, or $10^{\circ}C$, respectively. The acrylamide content of chips prepared from potatoes stored at $10^{\circ}C$ or $20^{\circ}C$ for one month was analyzed and the different frying times were 2, 3, 5, and 7 min. CONCLUSION: This study showed that monitoring and controlling the phosphate content within a potato field should be sufficient to avoid producing brown or black potato chips. For potatoes stored at low temperatures, a reconditioning period ($20^{\circ}C$ for 20 days) is required in order to reduce the levels of reducing sugars in the potato and subsequently reduce the acrylamide and improve chip coloration and appearance.

Characteristics of Whey Protein (WPC-30) Hydrolysate from Cheese Whey (치즈유청으로부터 제조한 유청단백질 가수분해물의 특성에 관한 연구)

  • Yoon, Yoh-Chang;An, Sung-Il;Jeong, A-Ram;Han, Song-Ee;Kim, Myeong-Hee;Lee, Chang-Kwon
    • Journal of Animal Science and Technology
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    • v.52 no.5
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    • pp.435-440
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    • 2010
  • Whey protein concentrate (WPC) is widely used to increase the nutritional and functional properties of food. In this study, the physiochemical and functionality of WPC-30 hydrolysates were examined to evaluate the possibility of application in the food industry. The WPC-30 was manufactured using ultrafiltration and spray-drying, and then hydrolyzed with proteolytic enzyme including alcalase, flavourzyme, nuetrase and protamex. Enzymatic hydrolysis had a significant influence on the physicochemical properties as evident from the increased foaming capacity, solubility. Alcalase caused highest protein hydrolysis (3.26%) and the bitterness. Foaming capacity was largest in WPC-30 hydrolysate treated with flavourzyme. Protein solubility at various levels of pH was highest in protamex-treated WPC-30 hydrolysate. However, the solubility of WPC-30 hydrolysates was significantly improved in alkaline condition than in acidic and neutral conditions. The study revealed that spray dried enzyme modified WPC can be used in various functional food.