• Title/Summary/Keyword: Lowry method

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Optimization of the Lowry Method of Protein Precipitation from the H. influenzae Type b Conjugate Vaccine Using Deoxycholic Acid and Hydrochloric Acid

  • Kim, Hyun-Sung;Kim, Sang-Joon;Kim, Hui-Jung;Kim, Han-Uk;Ahn, Sang-Joem;Hur, Byung-Ki
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.11 no.3
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    • pp.215-222
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    • 2006
  • The Lowry method was used in this study to measure protein in Haemophilus influenzae type b (Hib) conjugate vaccines (polyribosylibitol phosphate-tetanus toxoid; PRP-TT) using deoxycholic acid (DOC) to induce protein precipitation. Trichloroacetic acid (TCA) did not induce precipitation adequately from the Hib conjugate bulk and the freeze-dried Hib conjugate product. Its yield was approximately 50%. The matrix structure of Hib conjugate inhibits precipitation by TCA. Although the Lowry method can be carried out without precipitation in Hib conjugate bulk when no residual impurities (adipic acid dihydrazide [ADH], 1-ethyl-3-(3-dimethylamino-propyl) carbodiimide-HCI [EDAC], phenol and cyanogens bromide [CNBr], etc.) are present, it cannot be used for Hib conjugate products that contain sucrose 8.5%, because 8.5% concentration of sucrose enhanced the protein concentration. DOC- and HCl-induced precipitation is an alternative method for evaluating the protein content of the Hib conjugate bulk and the Hib conjugate product. The precipitation was optimal with a final concentrate of 0.1% for DOC at $4^{\circ}C$ and pH 2. This Lowry method, using DOC/HCI precipitation to induce protein precipitation, was confirmed a consistent, reproducible, and valid test for proteins in Hib conjugate bulk and its freeze-dried product.

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.

The Study on Protein Separations Using Ultrafiltration Membranes (한외여과막을 이용한 단백질 분리에 관한 연구)

  • 최원영;임균택이규현박돈희
    • KSBB Journal
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    • v.6 no.2
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    • pp.215-222
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    • 1991
  • The experimental investigations about ultrafiltration(UF) of bovine serum albumin(BSA) solutions were conducted with a batch stirred cell. This study was investigated on the permeate flux. The experiment was carried out at pH 4.8. Protein concentrations were measured spectrophotometrically. For very dilute samples, a Lowry method was used. In the ultrafiltration of dilute BSA solutions, the permeate flux according to working time was decreased smoothly and at isoelectric point, concentration polarization of BSA solution was ignored at limited working pressures by magnetic stirring.

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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.

Biochemical Changes in the Hemolymph of the Larvae of Thecodiplosis japonensis Uchi. et Inouye (솔잎혹파리 유충 체액의 생화학적 변화)

  • Lee Kyung-Ro;Lee Jong-Jin
    • Korean journal of applied entomology
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    • v.15 no.4 s.29
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    • pp.169-178
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    • 1976
  • The concentration of amino acids, total nitrogen, trehalose, lipids and the activities of respiratory, acid$\cdot$alkaline phosphatase, glutamic oxalozcetic transaminase and glutamic pyruvic transaminase during larval stage in Pine leaf gall midge, Thecodiplosis janensis Uchi. et Inouye were measured using Paper chromatographic method, micro-Kjeldahl method, Thin layer chromatographic method, Warburg's manometric method, Bessey-Lowry method and Reitman-Frankel method, respectively. Healthy specimens )yore chosen as samples of each larval stages; alrva in gall and larva in soil. Amino acids present in the alcoholic extracts were alanine, glutamic acid, glycine, histidine, methionine, proline, threonine, tryptophan and valine. The total nitrogen concentration reached to 31.348mg/g during the larva in gall and the larval stage in soil of the value was decreased to 29.027mg/g. The hemolymph sugar, trehalose value for larva in soil was about two times of the value for larva in gall. Total lipid, phospholipid,monoacylglycerol, triacylglycerol, sterol, free fatty acid and ester cholesterol were identified at larval stages in gall and soil. Triacylglycerol concentration reached high level in contrast with other lipid contents during larvae in gall and larva in soil. Free fatty acid, sterol except decreased lipids during larval stage in soil. Endogenous respiration, succinate of respiratory activities decreased at larval stage in soil compare with larva in gall. The activities of acid phosphatase decreased larval stage in soil but the activities of alkaline phosphatase increased remarkably. The activities of glutamic oxaloacetic transaminase and glutamic pyruvic transaminase reached high level of the larva in gall.

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A Study on Major Components of Bee Venom Using Electrophoresis (전기영동법(Electrophoresis)을 이용한 봉약침의 주요 성분에 관한 연구)

  • Lee, Jin-Seon;Kwon, Gi-Rok;Lee, Seung-Bae
    • Journal of Pharmacopuncture
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    • v.3 no.2
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    • pp.153-168
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    • 2000
  • This study was designed to study on major components of various Bee Venom(Bee Venom by electrical stimulation in Korea; K-BV I, Bee Venom by Microwave stimulation in Korea; K -BV II, 0.5rng/ml, Fu Yu Pharmaceutical Factory, China; C-BV, 1mg /ml, Monmouth Pain Institute, Inc., U.S.A.; A-BV) using Electrophoresis. The results were summarized as follows: 1. In 1:4000 Bee Venom solution rate, the band was not displayed distinctly usmg Electrophoresis. But in 1: 1000, the band showed clearly. 2. The results of Electrophoresis at solution rate 1:1000, K-BV I and K-BVII showed similar band. 3. The molecular weight of Phospholipase $A_2$ was known as 19,000 but its band was seen at 17,000 in Electrophoresis. 4. Protein concentration of Bee Venom by Lowry method was different at solution rate 1:4000 ; C-BV was $250{wmu}g/ml,\;K-BV\;I\;was\;190{wmu}g/ml,\;K-BVII\;was\;160{wmu}/ml\;and\;C-BV\;was\;45{wmu}/ml5$. Electrophoresis method was unuseful for analysis of Bee Venom when solution rate is above 1:4000 but Protein concentration of Bee Venom by Lowry method was possible. These data from the study can be applied to establish the standard measurement of Bee Venom and prevent pure bee venom from mixing of another components. I think it is desirable to study more about safety of Bee Venom as time goes by.

Studies on Antitumor Components of Wild Pholiota squarrosa (Fr.) Quel. (야생 비늘 버섯의 항암 성분에 관한 연구)

  • 박완희
    • YAKHAK HOEJI
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    • v.26 no.3
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    • pp.185-188
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    • 1982
  • In order to investigate the antitumor components of Korean wild higher fungi, the carpo-phores of Pholiota squarrosa collected in Kaung Nung area were extracted with water by refluxing for eight hours at 80-90.deg. C. The extracts were dialized for six days against distilled water and lyophilized to produce crude polysaccharide powders. They were found to have the antitumor activity against sarcoma 180 implanted in mice. Especially, the inhibition ratio of the extract of Pholiota squarrosa was 78.7% in the dose of 20mg/kg/day for the period of ten days. The tumor in two of the ten mice was completely regressed. The components of these aqueous extracts were found to be polysaccharide and protein. The total polysaccharide was 42% by Anthrone method and the protein was 55% by Lowry-Folin method in the antitumor component of Pholiota squarrosa.

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Discrimination of Native Bee-Honey and Foreign Bee-Honey by SDS-PAGE (단백질 전기영동을 이용한 토종꿀의 판별)

  • Lee, Deug-Chan;Lee, Sang-Young;Cha, Sang-Hoon;Choi, Yong-Soon;Rhee, Hae-Ik
    • Korean Journal of Food Science and Technology
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    • v.30 no.1
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    • pp.1-5
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    • 1998
  • To find out the difference between native bee-honey (NBH) and foreign bee-honey (FBH), quantification of honey protein and investigation of specific protein in NBH were carried out by SDS-PAGE. Contents of honey protein in NBH and FBH were measured by Bradford and Lowry method. The contents of protein determined by Bradford method were $0.1{\sim}3.3\;mg/g$ in NBH and $0.2{\sim}1.6\;mg/g$ in FBH, and by Lowry method were $12.9{\sim}45.7\;mg/g$ in NBH and $15.8{\sim}27.1\;mg/g$ in FBH. In order to investigate the distribution of bee honey proteins, the SDS-PAGE was performed. The results showed that molecular weight of the major proteins in NBH and in FBH were 56 kDa and 59 kDa. respectively. Therefore, it was confirmed that the difference between NBH and FBH can be identified visually by SDS-PAGE analysis. The major proteins in NBH and FBH were purified through two step chromatography, and the obtained proteins were used as marker protein in SDS-PAGE to discriminate NBH and FBH.

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Antitumor Components of Cryptoporus volvatus (한입버섯의 항암성분(抗癌成分)에 관한 연구(硏究))

  • Kim, Byong-Kak;Robbers, James E.;Chung, Kyeong-Soo;Chung, Hee-Soo;Choi, Eung-Chil
    • The Korean Journal of Mycology
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    • v.10 no.3
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    • pp.111-117
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    • 1982
  • The carpophores of Cryptoporus volvatus collected in Gyeong-gi Province of Korea were extracted with water and a protein-polysaccharide fraction was obtained after dialysis and lyophilization. The antitumor activity of this fraction was tested against sarcoma 180 implanted in A-strain mice. The tumor inhibition ratio was 80.4% in case of the high dose group (50mg/kg, ip, 10 days) and 70.3% in the low dose group (20mg/kg, ip, 10 days). The protein­polysaccharide fraction was chemically analyzed and was found to be a complex of a protein which was 18.2% of the fraction when determined by Lowry-Folin method, and a polysaccharide which was 55.3% of ther fraction when determined by Anthrone method. Their subunits were identified as four monosaccharides and 18 amino acids by gas-liquid chromatography and amino acid autoanalysis.

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Comparison of Non-saponin Composition and Contents in Fresh Ginseng Roots Cultivated in Different Areas and at Various Ages (수삼의 지역별 연근별 인삼 비사포닌 성분 함량 비교)

  • Yang, Byung-Wook;Im, Byung-Ok;Ko, Sung-Kwon
    • YAKHAK HOEJI
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    • v.50 no.4
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    • pp.215-219
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    • 2006
  • This study was carried out to obtain the basic information for non-saponin contents that can be used to index fresh ginseng roots (Panax ginseng C. A. Meyer) cultivated in the Republic of Korea and China. Non-saponin components in fresh gingeng roots which were cultivated in various areas and ages in Korea were determined. Acidic polysaccharide, total polysaccharide, crude polyacetylene were quantitatively analyzed by using the method of spectrophotometric determination, while the total protein was analyzed by using Lowry method. The results show that there were no statistically significant differences for the average contents of four non-saponins among 4-years-old, 5-years-old, and 6-years-old fresh ginseng roots. Additionally, this study assessed the average contents of non-saponin components in 4-years-old fresh ginseng roots (Panax ginseng C. A. Meyer) which were cultivated in Korea and China. The result showed that the average contents of crude polyacetylene and acidic polysaccharide were statistically significant. Four-years-old fresh ginseng roots cultivated in Korea had the higher average contents of crude polyacetylene and acidic polysaccharide than those cultivated in China. However the average contents of total polysaccharide and total protein had no statistically significant difference.