• Title/Summary/Keyword: Lowry protein assay

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Proteolysis of Defatted Rice Bran Using Commercial Proteases and Characterization of Its Hydrolysates (탈지미강 단백질의 가수분해 및 분해물의 특성 연구)

  • Kim, Chang-Won;Kim, Hyun-Seok;Kim, Byung-Yong;Baik, Moo-Yeol
    • Food Engineering Progress
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    • v.15 no.1
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    • pp.41-47
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    • 2011
  • The defatted rice bran (DRB) was enzymatically hydrolyzed using eight commercial proteases for 4hr at optimum pH and temperature. Proteolytic hydrolysates were examined in supernatant and precipitate using lowry, semimicro kjeldahl and gravimetric method using weight difference before and after enzymatic hydrolysis. In lowry and kjeldahl protein assay method, two proteases (Alcalase and Protease N) were found to be the most effective enzymes. In gravimetric method, 60.6~118.3 mg protein/g DRB was hydrolyzed after eight commercial proteases treatments. Similar to lowry and kjeldahl method, 118.3 and 107.1 mg protein/g DRB were hydrolyzed after Alcalase and Protease N treatments, respectively. When two or three effective proteases (Protamex, Alcalase and Protease N) were applied at one time to obtain synergistic effect, significant increase (P<0.05) was observed when three proteases were applied at one time (63.4 mg protein/g DRB in lowry method and 204.5 mg protein/g DRB in gravimetric method). This result suggests that Alcalase and Protease N were the most effective enzymes for proteolysis of DRB and three commercial enzymes (Protamex, Alcalase and Protease N) showed the synergistic effect on the hydrolysis of DRB.

Hydrolysis of Rice Syrup Meal Using Various Commercial Proteases (쌀 시럽박의 단백질 가수분해 특성)

  • Kim, Chang-Won;Park, Jin-Woo;Choi, Hyuk-Joon;Han, Bok-Kyung;Yoo, Seung-Seok;Kim, Byung-Yong;Baik, Moo-Yeol;Kim, Young-Rok
    • Journal of Life Science
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    • v.21 no.2
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    • pp.309-315
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    • 2011
  • Rice syrup meal (RSM) was enzymatically hydrolyzed using eight commercial proteases (Protamex, Neutrase, Flavourzyme, Alcalase, Protease M, Protease N, Protease A, Molsin F) for 4 hr at optimum pH and temperature. Proteolytic hydrolysates were examined in supernatant and precipitate using Lowry protein assay, semimicro Kjeldahl method and gravimetric method using weight difference before and after enzymatic hydrolysis. Although RSM contains a high amount of protein (71.2%), only a very small amount of protein was hydrolyzed. Two proteases (Protease M and Protease N) were found to be the most effective in the hydrolysis of RSM protein. In Lowry method, 57.5 and 59.0 mg protein/g RSM were hydrolyzed after Protease M and Protease N treatments, respectively. In gravimetric method, 80.0 and 85.4 mg protein/g RSM were hydrolyzed after Protease M and Protease N treatments. In Kjeldahl method, 67.43 and 70.43 mg protein/g RSM were hydrolyzed after Protamex and Protease N treatments, respectively. For synergistic effect, two or three effective commercial proteases (Protease M, Protease N and Protease A) were applied to RSM at one time. The highest hydrolysis of RSM protein was observed in both Lowry protein assay (80.3 mg protein/g RSM) and gravimetric methods (153.2 mg protein/g RSM) when three commercial proteases were applied at one time, suggesting the synergistic effect of those proteases.

Bio-Rex 젤을 이용한 난백으로 부터의 Lysozyme의 분리

  • Kim, Hyeong-Won;Park, In-Gyu;Song, Jae-Yang;Kim, In-Ho
    • 한국생물공학회:학술대회논문집
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    • 2002.04a
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    • pp.493-496
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    • 2002
  • Lysozyme is an enzyme which has the ability to lyse bacteria such as Micrococcus lysodeikticus or gram positive and gram negative bacteria by hydrolyzing in the peptidoglycan layer of the bacterial cell wall. Lysozyme is abundantly contained in an egg white. In order to obtain lysozyme from egg white, we used Bio-rex ion exchange chromatography and can identify the exist of lysozyme by SDS-PAGE and protein assay.

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Derivatization of Rice Wine Meal Using Commercial Proteases and Characterization of Its Hydrolysates (단백질가수분해효소를 이용한 탁주박의 가수분해 및 그 분해물의 특성)

  • Kim, Chang-Won;Choi, Hyuk-Joon;Han, Bok-Kyung;Yoo, Seung-Seok;Kim, Chang-Nam;Kim, Byung-Yong;Baik, Moo-Yeol
    • Korean Journal of Food Science and Technology
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    • v.43 no.6
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    • pp.729-734
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    • 2011
  • With the goal of transforming rice protein from an insoluble to a soluble form to increase the industrial utilization of rice wine meal (RWM), RWM was derivatized using commercial proteases and the RWM hydrolysates were characterized. Eight commercial proteases were used individually or in combination for hydrolysis of RWM. The degree of hydrolysis was assessed by determining the soluble protein in supernatant using the Lowry assay, protein in precipitates using a semimicro Kjeldahl procedure, and gravimetrically by the weight difference before and after hydrolysis. Protamex, Alcalase and Protease N proteases were most effective for hydrolysis of RWM. Although these assessment methodologies displayed some variation, they generally showed a similar pattern. When the aforementioned three proteases were simultaneously used to treat RWM, no significant difference was observed between the three assays (p<0.05) indicating an absence of enzymatic synergy.

The Difference of the Cleaning and Wettability-maintaining Efficacy of Lens Care Solution to RGP Lens (관리 용품에 따른 RGP 렌즈의 세척효과 및 습윤성 차이)

  • Kim, Myoung-Hea;Park, Mi-Jung
    • Journal of Korean Ophthalmic Optics Society
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    • v.11 no.1
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    • pp.27-34
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    • 2006
  • We investigated the question whether the efficacy of cleaning tear components on RGP lens and preserving the superior wettability of RGP lens depended on the different type of contact lens care system - RGP lens care solution, SCL care solution, combined solution both for SCL and RGP lens or saline solution. The removal efficacy of the deposited protein was examined by Lowry protein assay and Scanning Electro Microscope(SEM) and residual lipid concentration on RGP lens was determined by High Pressure Liquid Chromatology(HPLC). Wettability was assessed with an equilibrium water-in-air contact angle method. When cared by RGP lens solution, it was demonstrated that 62 percent out of the adhered protein on RGP lens were removed and the removal efficacy of RGP lens solution was not only 4 times than saline solution and the alternative but also higher twice than SCL solution. Contrarily, the SCL solution had the most excellent removal efficacy of the adhered protein on SCL. These results suggest that the cleaning efficacy is thought to be affected by the other factors like the viscosity of care solutions, which mutual contact between RGP lens and care solutions is on the increase due to the viscosity enhancer in RGP lens care solution. RGP lens solution had the greatest removing efficacy to cholesterol and the residual cholesterol concentration was decreased to 50%. It is significant for RGP lens to preserve the superior wettability which means the predictive value for comfortable wearing and it showed that the RGP lens solution offered the most excellent efficacy to maintain the surface wettability. Combined solution both for SCL and RGP lens had weak efficacy of cleaning and maintaining wettability for RGP lens compared to RGP lens care solution.

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A Study on The Content of Liver Protein, Nucleic Acids, and Guanine Deaminase Activity of Mouse During Acute Starvation (급성(急性) 기아(饑餓)마우스의 간단백질(肝蛋白質), 핵산(核酸) 및 Guanine Deaminase 활성(活性)에 관(關)한 연구(硏究))

  • Park, Seung-Hee;Kim, Seung-Won
    • Journal of Nutrition and Health
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    • v.1 no.2
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    • pp.107-115
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    • 1968
  • Number of aspects, not only nutritional but social as well as political involved in human starvation pose nowadays global problems. In order to help establish the minimum nutritional requirements in the daily life of a man and to free people as well from either undernourishment, malnutrition or even starvation many workers have devoted themselves so far on the research programs to know what and how number of metabolic events take place in animals in vivo. It is the purpose of the present paper to examine in effect to what extent both of the protein and nucleic acids (DNA & RNA) together with an enzyme, guanine deaminase, which converts guanine into xanthine and in turn ends up to uric acid as an end product, undergo changes, quantitatively during acute starvation, using the mouse as an experimental animal. The mouse was strictly inhibited from taking foods except drinking water ad libitum and was sacriflced 24, 48, and 72 hours following starvation thus acutely induced. The animals consisted of two experimental groups, one control and another starvation groups, each being consisted of 6-24 mice of whose body weights ranged in the vicinity of 10 g. The animals were sacriflced by a blow on the head, followed by immediate excision of their livers into ice-cold distilled water, washing adherent blood and other contaminant tissues. The liver was minced foramin, by an all-glass homogenizer immersing it in an ice-bath, followed by subsequent fractionatin of the homogenate (10% W/V in 0.25M sucrose solution made up with 0.05M phosphate buffer of pH 7.4). For the liver protein and guanine deaminase assay, the 10% homogenate was centrifuged at 600 x g for 10 minutes to eliminate the nuclear fraction; and for the estimation of DNA and RNA, the homogenate was prepared by the addition of 10% trichloroacetic acid in order to free the homogenate from the acid-soluble fraction, the remaining residue being delipidate by the addition of alcohol and dried in vacuo for later KOH (IN) hydrolysis. The changes in body and liver wegihts during acute starvation were checked gravimetrically. Protein contents in the liver were monitored by the method of Lowry et al; and guanine deaminase activities were followed by the assay of liberated ammonia from the substrate utilizing the Caraway's colorimetry. The extraction of both DNA and RNA was performed by the Schmidt-Thannhauser's method, which was followed by Marmur's method of purification for DNA and by Chargaff's method of purification for RNA. The determinations of both DNA and RNA were carried out by the diphenylamine reaction for the former and by the orcinol reaction for the latter. The following resume was the results of the present work. 1. It was observed that the body as well as liver weights fall abruptly during starvation, and that the loss of body weight showed no statistical correlation with the decreases in the content of liver protein. 2. The content of liver protein and activity of liver guanine deaminase activity as well decline dramatically, and the specific activities of the enzyme (activity/protein), however, decreased gradually as starvation proceeded. 3. Both of the nucleic acids, DNA and RNA, showed decrements in the liver of mouse during acute starvation; the latter, however, being more striking in the decline as compared to the former. 4. The decreases in the liver protein content as resulted from the acute starvation had no statistically significant correlation with the decrements of DNA in the same tissue, but had regressed with a significant statistical correlation with the fall of RNA in the tissue. 5. The decrease in the activity of guanine deaminase in the liver of mouse during acute starvation was functionally more proportional to the decrease in RNA than DNA, and moreover correlated with the changes in the content of the liver protein. 6. The possible mechanisms involved during in this acute starvation as bring the decreases in the contents of DNA, protein, and guanine deaminase were discussed briefly.

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