• Title/Summary/Keyword: crude extracts

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Enzymatic Studies on the α-Galactosidases from Soybean and Aspergillus niger (대두(大豆) 및 Aspergillus niger α-galactosidase의 효소학적(酵素學的) 연구(硏究))

  • Keum, Jong-Hwa;Oh, Man-Jin
    • Korean Journal of Agricultural Science
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    • v.18 no.1
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    • pp.49-73
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    • 1991
  • To elucidate enzymatic properties of $\alpha$-galactosidases (EC3, 2, 1, 22) from germinated soybean and Aspergillus niger changes in the enzyme activities and oligosaccharide contents during germination of soybean were determined and $\alpha$-galactosidases from germinated soybean and wheat bran culture of Aspergillus niger were purified by ammonium sulfate fractionation, ion exchange chromatography and gel filtration. Their chemical and enzymatic properties were investigated and the results obtained were summarized as follows : 1. $\alpha$-Galactosidase activity of soybean was maximized when it was germinated at $25^{\circ}C$ for 120 hours. And raffinose and stachyose in soybean were decomposed completely after 96 hours and 120 hours of germination, respectively. 2. The highest level of $\alpha$-Galactosidase activity was obtained when Aspergillus niger was grown on wheat bran medium at $30^{\circ}C$ for 96 hours. 3. Soybean $\alpha$-galactosidase was purified by 6.6 fold by ammonium slufate fractionation, ion exchange chromatography on DEAE-Cellulose and Sephadex A-50., and gel filtration on Sephadex G-150. Its specific activity was 825 units/mg protein and the yield was 2.5% of the total activity of crude extracts. 4. Aspergillus niger $\alpha$-galactosidase was purified by 23.7 fold. Its specific activity was 1,229 units/mg protein and the yield was 14% of the total activity of wheat bran culture. 5. The purified $\alpha$-galactosidases of soybean and Aspergillus niger were found to be homogeneous by polyacrylamide gel electrophoresis and by HPLC. 6. Chemical properties of the purified $\alpha$-galactosidases were : 1) The soybean $\alpha$-galactosidase was monomeric and its molecular weight was estimated to be 30,000 by SDS-PAGE whereas the Aspergillus niger $\alpha$-galactosidase was a tetrameric glycoprotein which consisted of identical subunits with molecular weight of 28,000 each.

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Purification and Characterization of a Lipolytic Enzyme Produced by Aeromonas hydrophila PL43 (Aeromonas hydrophila PL43이 생산하는 지질분해 효소의 정제 및 특성)

  • Kim, Yong-Woo;Hong, Sung Wook;Chung, Kun Sub
    • Microbiology and Biotechnology Letters
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    • v.44 no.2
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    • pp.130-139
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    • 2016
  • A bacterial strain, producing an excellent lipolytic enzyme, was isolated from the intestinal tracts of an earthworm (Eisenia fetida). The strain was identified as Aeromonas hydrophila by phenotypic, chemotaxonomic characteristics and 16S ribosomal DNA analysis, and was designated as Aeromona hydrophila PL43. The lipolytic enzyme from A. hydrophila PL43 was purified via 35−45% ammonium sulfate precipitation, DEAE-sepharose fast flow ion-exchange, and sephacryl S-300HR gel filtration chromatography. The yield of the purified enzyme was 3.7% and 2.5% of the total activity of crude extracts with p-nitrophenyl butyrate (pNPB) and p-nitrophenyl palmitate (pNPP) as substrates, respectively. The molecular weight of the purified enzyme was approximately 74 kDa using gel filtration, sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and zymography. The optimal activity of purified enzyme was observed at 50℃ and pH 8.0 using pNPB, and 60℃ and pH 8.0 using pNPP. The purified enzyme was stable in the ranges 20− 60℃ and pH 7.0−10.0. The activity of purified enzyme was inhibited by PMSF, pepstatin A, Co2+, Cu2+, and Fe2+, but was recovered by metal chelating of EDTA. The Km and Vmax values of the purified enzyme were 1.07 mM and 7.27 mM/min using pNPB and 1.43 mM and 2.72 mM/min using pNPP, respectively.

Identification of a New Potyvirus, Keunjorong mosaic virus in Cynanchum wilfordii and C. auriculatum (큰조롱과 넓은잎 큰조롱에서 신종 포티바이러스(큰조롱모자이크바이러스)의 동정)

  • Lee, Joo-Hee;Park, Seok-Jin;Nam, Moon;Kim, Min-Ja;Lee, Jae-Bong;Sohn, Hyoung-Rac;Choi, Hong-Soo;Kim, Jeong-Soo;Lee, Jun-Seong;Moon, Jae-Sun;Lee, Su-Heon
    • Research in Plant Disease
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    • v.16 no.3
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    • pp.238-246
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    • 2010
  • In 2006 fall, a preliminary survey of viruses in two important medicinal plants, Cynanchum wilfordii and C. auriculatum, was conducted on the experimental fields at the Agricultural Research and Extension Services of Chungbuk province in Korea. On each experimental fields, percentage of virus infection was ranged from 20 to 80%, and especially an average of disease incidence propagated by roots was twice higher than that by seeds. The various symptoms were observed in Cynanchum spp. plants, such as mosaic, mottle, necrosis, yellowing, chlorotic spot and malformation etc. In electron microscopic examination of crude sap extracts, filamentous rod particles with 390-730 nm were observed in most samples. The virus particles were purified from the leaves of C. wilfordii with typical mosaic symptom, and the viral RNA was extracted from this sample containing 430-845 nm long filamentous rod. To identify the viruses, reverse transcription followed by PCR with random primers was carried out. The putative sequences of P3 and coat protein of potyvirus were obtained. From a BLAST of the two sequences, they showed 26-38% and 62-72% identities to potyviruses, respectively. In SDS-PAGE analysis, the subunit of coat protein was approximately 30.3 kDa, close to the coat protein of potyvirus. In bioassay with 21 species in 7 families, Chenopodium quinoa showed local lesion on inoculated leave and chlorotic spot on upper leave, but the others were not infected. RT-PCR detection using specific primer of C. wilfordii and C. auriculatum samples, all of 24 samples with virus symptom was positive, and five out of seven samples without virus symptom were also positive. On the basis of these data, the virus could be considered as a new member of potyvirus. We suggested that the name of the virus was Keunjorong mosaic virus (KjMV) after the common Korean name of C. wilfordii.

Antimicrobial Activity of Bamboo (Sasa borealis) Leaves Fraction Extracts against Food Poisoning Bacteria (조릿대 잎 분획 추출물의 식중독균에 대한 항균활성)

  • Park, Yeon-Ok;Lim, Hyeon-Sook
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.39 no.12
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    • pp.1745-1752
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    • 2010
  • This study was conducted to investigate the antimicrobial activity of 70% ethanol (EtOH) extract and the five fractions of the crude extract from Sasa borealis leaves against seven food poisoning bacteria, Staphylococcus aureus, Micrococus luteus, Listeria monocytogens, Bacillus subtilis, Escherichia coli, Salmonella Typhimurium, and Pseudomonas aeruginosa. The yield of 70% EtOH extract was 11.4% and those of n-hexane, chloroform, ethyl acetate, n-butanol, and aqueous fractions were 3.0%, 1.1%, 0.6%, 1.3%, and 5.1%, respectively. The 70% EtOH extract and the four fractions except aqueous fraction demonstrated antimicrobial activity against all the seven food poisoning bacteria at a concentration of 0.5%, although it was less compared to benzoic acid. Minimal inhibitory concentration (MIC) of the 70% EtOH extract against all the food poisoning bacteria except S. aureus was $50{\mu}L$/disc. Moreover, chloroform fraction was $35{\mu}L$/disc against 3 food poisoning bacteria and $50{\mu}L$/disc against the other 4 food poisoning bacteria; ethyl acetate fraction was $50{\mu}L$/disc against all the food poisoning bacteria. In addition, n-butanol fraction was $50{\mu}L$/disc against all the food poisoning bacteria except S. aureus. Aqueous fraction, which did not show antimicrobial activity at 5%, was $200{\mu}L$/disc against only S. aureus and L. monocytogen. The 0.25%, and 0.5% of ethyl acetate fraction inhibited the growth of all the food poisoning bacteria 8 to 12 hours and 24 hours, respectively. These results indicate that the Sasa borealis leaves may be useful as a natural antimicrobial substance.

Antioxidant and Anti-Inflammatory Activities of Eugenol and Its Derivatives from Clove (Eugenia caryophyllata Thunb.) (정향(Eugenia caryophyllata Thunb.) Eugenol 및 그 유도체의 항산화 및 항염증활성)

  • Leem, Hyun-Hee;Kim, Eun-Ok;Seo, Mi-Jae;Choi, Sang-Won
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.40 no.10
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    • pp.1361-1370
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    • 2011
  • Antioxidant and anti-inflammatory activities of eugenol and its derivatives from clove (Eugenia caryophyllata Thunb.) were evaluated using in vitro assay systems by measuring 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical, cyclooxygenase-2 (COX-2), and 15-lipoxygenase (15-LOX). Among eight different crude medicinal drugs tested, volatile extracts of clove extracted by steam distillation extraction (SDE) showed potent DPPH radical scavenging activity ($IC_{50}$=8.85 ${\mu}g/mL$) as well as strong inhibitory activity against COX-2 (58.15%) and 15-LOX (86.15%) at 10 ${\mu}g/mL$ and 25 ${\mu}g/mL$, respectively. Major volatile components of clove were identified as eugenol, trans-caryophyllene, and acetyleugenol by GC-MS analysis. Out of three eugenol derivatives, eugenol, methyl eugenol, and acetyl eugenol, eugenol showed the strongest DPPH radical scavenging activity and COX-2 inhibitory activity, whereas methyl eugenol exhibited the strongest 15-LOX inhibitory activity. Finally, the contents of the three eugenol derivatives in clove were quantified by analytical HPLC. Contents of eugenol and acetyl eugenol in clove were 6.95% and 1.85% per dry weight, respectively. These results suggest that eugenol and its derivatives in steam distilled extract of clove may be useful as potential antioxidant and anti-inflammatory agents.

Chemical Composition and Physiological Activity of Opuntia ficus-indica depending on Different Cultivation Regions (재배지역별 보검선인장 줄기의 영양성분 및 생리활성 평가)

  • Lee, Sang Hoon;Jeong, Yun Sook;Song, Jin;Hwang, Kyung-A;Noh, Geon Min;Hwang, In Guk
    • The Korean Journal of Food And Nutrition
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    • v.29 no.4
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    • pp.521-528
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    • 2016
  • This study was conducted to investigate changes in the proximate composition, antioxidant activities, and ${\alpha}$-glucosidase inhibitory activity of Opuntia ficus-indica (OFI) cladodes cultivated in Jeju (JJ1, JJ2, JJ3) and Jeonnam (JN1, JN2). The difference in the proximate composition (crude protein, lipid and ash content) of OFI between the two regions was not significant. Ca, Mg and Na were the major mineral components of OFI. The ascorbic acid content of OFI ranged from 57.87 to 143.72 mg/100 g. A 70% ethanol extract was used to investigate the antioxidant content and activity as well as the ${\alpha}$-glucosidase inhibitory activity. The total polyphenol and flavonoid contents of OFI were 38.69~55.29 and 3.33~4.03 mg/g, respectively. The antioxidant activities based on the DPPH and ABTS free radical scavenging assays were 45.19~61.52% and 39.15~51.96%, respectively, at a concentration of 1 mg/mL. The inhibitory activity of OFI extracts against rat intestinal ${\alpha}$-glucosidase was 29.72~45.73% at 1 mg/mL concentration, and JN1 showed the highest ${\alpha}$-glucosidase inhibitory activity. This information could be very useful for authentication of Opuntia species with the highest potential as sources of nutritional and therapeutic elements.

Comparison of Lipid Content and Monounsaturated Fatty Acid Composition of Beef by Country of Origin and Marbling Score (원산지와 근내지방도에 따른 소고기의 지방 함량과 단일불포화지방산 조성 비교)

  • Kwon, Ha Na;Choi, Chang Bon
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.44 no.12
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    • pp.1806-1812
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    • 2015
  • This study was conducted to compare moisture, crude protein, ether extracts, fatty acid composition, and melting point of beef with different origins. Eighty (80) samples of domestic Hanwoo beef with different marbling scores (BMS) of 3, 5, 7, and 9, respectively (20 samples for each BMS) and 30 samples of imported beef (15 samples each of American and Australian beef) were used. Relationship of fatty acid composition with melting point of lipids was also analyzed. Percentages of ether extract of Hanwoo beef with BMS of 3, 5, 7, and 9 as well as American and Australian beef were 11.91, 13.23, 17.03, 23.92, 8.36, and 4.47%, respectively, with the highest value in Hanwoo with BMS of 9 and lowest value in Australian beef. Percentages of oleic acid (C18:1n9), the most abundant monounsaturated fatty acid (MUFA) in beef, were 44.92, 47.19, 46.23, 47.70, 43.70, and 38.28%. MUFA/saturated fatty acid (SFA) (M/S) ratios were 1.15, 1.25, 1.22, 1.27, 1.10, and 0.86, respectively. The melting point of lipids extracted from beef samples was the lowest ($27.42^{\circ}C$) in Hanwoo with BMS of 9, whereas it was highest ($41.15^{\circ}C$) in Australian beef. SFAs such as myristic acid (C14:0), palmitic acid (C16:0), stearic acid (C18:0), and total SFA in beef samples showed positive (+) correlations (r=0.203, 0.279, 0.807, and 0.880, respectively) with melting points, whereas MUFAs, palmitoleic acid (C16:1), oleic acid (C18:1n9), and total MUFA, showed negative (-) correlations (r=-0.541, -0.857, and -0.906, respectively). In conclusion, as the lipid contents of beef increased, composition of oleic acid (C18:1n9) increased. Furthermore, as BMS increased in Hanwoo beef, M/S was increased.

Effects of Peonia Seed Extracts and Resveratrol on Lipid Metabolism in Rats Fed High Cholesterol Diets (작약(Paeonia lactiflora Pall.)씨 추출물과 Resveratrol이 흰쥐 체내 지질 상태에 미치는 영향)

  • 서상희;이향림;이순재;최상원;조성희
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.32 no.7
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    • pp.1102-1107
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    • 2003
  • To study the effects of substances in paeonia seeds (Paeonia lactiflora Pall.) on lipid metabolism, crude methanol extract and secondary ether-soluble fraction out of defatted methanol extract and trans-resveratrol were prepared from the seeds and added to 0.5% (w/w) cholesterol diets for rats. Male Sprague-Dawley rats weighing 120$\pm$11 g were divided into six experimental groups: control group with no extra supplement, 0.1% (MP1) and 0.2% methanol extract (MP2) supplemented groups, 0.05% (EP1) and 0.1% ether-soluble fraction (EP2) supplemented groups and 0.02% resveratrol supplemented group. Experimental diets were fed ad libitum to the rats for 3 weeks. Body weight gains and food efficiencies were not different among the six experimental groups. Relative liver weights were lower in EP2 group compared to those in control group, but serum GOT and GPT levels of paeonia seed groups including trans-resveratrol group were not different from those of the control group. Serum total cholesterol levels reduced in EP2 and resveratrol groups but HDL-/total cholesterol ratios significantly increased in the four paeonia groups except EP1 group compared with the control group and serum triglyceride level lowered only in EP2 group than that of the control. However, liver cholesterol levels lowered in the five paeonia groups but triglyceride level lowered in MP2, EP1, EP2 groups than that of the control group. Fecal cholesterol excretion significantly increased in MP2, EP1, and EP2 groups than that of the control group, but bile acid excretions were not changed except that a reduction in EP2 group. These results suggest that paeonia seeds contain substances improving serum lipid status mostly via HDL pathway and resveratrol as monomer is one of the effective components but others including resveratrol oligmer are involved in the lipid improving effect.

Changes in Antioxidant Contents and Activities of Adzuki Beans according to Germination Time (발아시간에 따른 팥의 항산화성분 및 항산화활성의 변화)

  • Woo, Koan Sik;Song, Seuk Bo;Ko, Jee Yeon;Lee, Jae Saeng;Jung, Tae Wook;Jeong, Heon Sang
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.44 no.5
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    • pp.687-694
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    • 2015
  • The purpose of this study was to evaluate the yield, antioxidant content, and antioxidant activity of adzuki beans according to germination time. Cultivated varieties were Vigna angularis var. Nipponensis cv. Chungju-pat (CJP), and Yeonduchae (YDC), and Vigna radiata (L.) R. Wilczek cv. Dahyeon (DH). The moisture, crude protein, calcium, and magnesium contents of YDC significantly changed with increasing germination time, whereas potassium, natrium, and aluminium contents did not significantly change. Sprout yield, total polyphenol, flavonoid, and tannin contents of ethanolic extracts from adzuki and mung beans significantly increased with increasing germination time. Total polyphenol contents of ungerminated CJP, YDC, and DH were 1.96, 2.68, and 2.02 mg/g, and those of CJP and YDC germinated for 144 h were 3.33 and 3.47 mg/g, respectively. Total flavonoid content of adzuki beans substantially decreased with increasing germination time. Total tannin content substantially increased with increasing germination time, and YDC showed higher contents (0.85 mg/g) sample germinated for 120 h. DPPH radical scavenging activities of CJP and YDC substantially decreased with increasing germination time, whereas that of DH increased. ABTS radical scavenging activities of ungerminated CJP, YDC, and DH were 4.57, 6.51, and 2.82 mg/g, respectively, and increased after germination for 72~120 h.

Investigation of radical scavenging and anti-inflammatory activity of the extract and fractions of Termitomyces albuminosus (계종버섯 추출물과 분획물의 라디칼 소거 활성과 항염증 활성 분석)

  • Kang, Min-Jae;Kim, Young-Jae;Hwang, Hyeon-Jeong;Kim, Moon-Ho;Sin, Jong-Moo;Shin, Pyung-Gyun;Kim, Gun-Do;Kim, Keun Ki
    • Journal of Mushroom
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    • v.18 no.1
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    • pp.45-52
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    • 2020
  • Termitomyces albuminosus has been recognized to have the best mushrooms in China, in terms of taste and aroma. The efficacy of these mushrooms has been recorded in the botanical list. However, research on the development of their artificial culture methods is necessary. In this study, we prepared an organic solvent extract and a hot water extract to understand the development of compounds and functional foods with antioxidant and anti-inflammatory activities. The IC50 value of DPPH radical scavenging activity of the hot water extract (TA4) was 1.5 mg/mL and the IC50 value of the MeOH fraction (TA2) was 1.93 mg/mL. The anti-inflammatory activity was investigated by the inhibition of NO production. EtOAc fraction (TA1) is a crude extract, but 79% of NO production was inhibited at 100 ㎍/mL. NO was not produced at 200 ㎍/mL. TA1-5-6, from TA1 inhibited NO production by 15% as compared to the positive control at 15 ㎍/mL, and completely inhibited NO production at 30 ㎍/mL. No cytotoxicity was observed at 50 ㎍/mL. TA2-1-5 from the MeOH fraction (TA2) inhibited more than 75% of NO production at 30 ㎍/mL; cytotoxicity was very low even at 50 ㎍/mL. In conclusion, by selective solvent selection, it was possible to manufacture an extract with no cytotoxicity and excellent biological activities. Furthermore, the extracts showed potential for developing various functional foods and drugs.