• Title/Summary/Keyword: Extraction yield

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High Yield Bacterial Expression and Purification of Active Cytochrome P450 p-coumarate-3-hydroxylase (C3H), the Arabidopsis Membrane Protein (대장균 시스템을 이용한 Arabidopsis 막 단백질 cytochrome P450 p-coumarate-3hydroxylase (C3H) 활성형의 과발현 및 분리정제)

  • Yang, Hee-Jung;Kim, Wan-Yeon;Yun, Young-Ju;Yoon, Ji-Won;Kwon, Tae-Woo;Youn, Hye-Sook;Youn, Bu-Hyun
    • Journal of Life Science
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    • v.19 no.8
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    • pp.1039-1046
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    • 2009
  • The cytochrome P450s (P450s) metabolizing natural products are among the most versatile biological catalysts known in plants, but knowledge of the structural basis for their broad substrate specificity has been limited. The activity of p-coumarate 3-hydroxylase (C3H) is thought to be essential for the biosynthesis of lignin and many other phenylpropanoid pathway products in plants however, all attempts to express and purify the protein corresponding C3H gene have failed. As a result, no conditions suitable for the unambiguous assay of the enzyme are known. The detailed understanding of the mechanism and substrate-specificity of C3Hdemands a method for the production of active protein on the milligram scale. We have developed a bacterial expression and purification system for the plant C3H, which allows for the quick expression and purification of active wild-type C3H via introduction of combinational mutagenesis. The modified cytochrome P450 C3H ($C3H_{mod}$) could be purified in the absence of detergent using immobilized metal affinity chromatography and size exclusion chromatography following extraction from isolated membranes in a high salt buffer and catalytically activated. This method makes the use of isotopic labeling of C3H for NMRstudies and X-ray crystallography practical, and is also applicable to other plant cytochrome P450 proteins.

Anti-oxidative and Neuroprotective Activities of Pig Skin Gelatin Hydrolysates (돈피젤라틴 효소분해물의 항산화 활성 및 신경세포보호효과)

  • Kim, Dong Wook;Park, Kimoon;Ha, Goeun;Jung, Ju Ri;Chang, Ounki;Ham, Jun-Sang;Jeong, Seok-Geun;Park, Beom-Young;Song, Jin;Jang, Aera
    • Food Science of Animal Resources
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    • v.33 no.2
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    • pp.258-267
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    • 2013
  • This study was conducted to determine the antioxidative and neuroprotective effect of pig skin extracts (PS) and pig skin gelatin hydrolysates (LPS) using a human neuroblastoma cell line (SH-SY5Y). The extraction yield of PS was 3 fold higher than that of LPS. The protein content of PS was about 10 fold higher than that of LPS (p<0.05). Also LPS increased antioxidative activity dose dependently, and the activity was significantly higher than PS at all concentration (p<0.05). DPPH radical scavenging activity of LPS at 50 mg/mL was 92.97%, which was similar to $1{\mu}M$ vitamin C as a positive control. ABTS radical scavenging activity of LPS (20 mg/mL) was 89.83% and oxygen radical absorbance capacity of LPS at 1 mg/mL was $141.39{\mu}M$ Trolox Equvalent/g. No significant change of human neuroblastoma cells was determined by MTT test. Cell death by oxidative stress induced by $H_2O_2$ and amyloid beta 1-42 ($A{\beta}_{1-42}$) was protected by LPS rather than PS. Acetylcholine esterase was significantly inhibited, by up to 33.62% by LPS at 10 mg/mL. Therefore, these results suggest that pig skin gelatin hydrolysates below 3 kDa have potential to be used as anti-oxidative and neuroprotective functional additives in the food industry, while further animal test should be determined in the future.

Lipid Characteristics of Fish Frame as a Functional Lipid Resource (기능성 지질 추출 소재로서 Fish Frames의 지질성분 특성)

  • Kim, Jeong-Gyun;Han, Byung-Wook;Kim, Hye-Sook;Park, Chan-Ho;Chung, In-Kwon;Choi, Yeung-Joon;Kim, Jin-Soo;Heu, Min-Soo
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.34 no.3
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    • pp.380-388
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    • 2005
  • Lipid characteristics of 6 species of fish frame (armored weasel-fish, AW; chum salmon, CS; spanish mackerel, SM; common mackerel, CM; conger eel, CE and skipjack tuna, ST) were studied by determining total lipid (TL) contents, lipid classes, and fatty acid composition. The highest yield of bone was obtained from ST frame (64.2%), followed by CS frame (57.9%), CE frame (54.6%), A W frame (41.6%), SM frame (41.7%), and CM frame (32.6%). The highest neutral lipid content was also found in total lipid (TL) from SM bone (23.3 g/100 g), followed by TL from CE bone (21.5 g/100 g), CS bone (16.0 g/100 g), and CM bone (15.5 g/100 g), while those from ST and A W bones were 7.2 g/100 g and 0.4 g/100 g, respectively. The prominent lipid classes of neutral lipids (NL) from all fish bones and muscles were triglyceride (TG), however, which was much lower in AW than in other fishes. The percentages of EPA and DHA in NL from fish bone were in the descending order of CS (29.3%), ST (27.1%), AW (27.0%), CM (25.7%), SM (21.6%), and CE (14.9%). Based on the lipid characteristics, the CS frame was the best resource for extraction of a functional lipid.

Characteristics of Hot-Water Extracts from Salmon Frame as Basic Ingredients for Gomtang-like Products (열수추출 연어 Frame 엑스분의 곰탕 유사 제품 베이스로서의 특성)

  • Han, Byung-Wook;Kim, Hye-Suk;Jee, Seung-Joon;Lee, Jae-Hyoung;Kim, Hyung-Jun;Park, Shin-Ho;Ji, Seong-Gil;Heu, Min-Soo;Kim, Jin-Soo
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.36 no.10
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    • pp.1326-1333
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    • 2007
  • For preparing the basic ingredients of Gomtang-like products from the extracts of salmon frame, the extraction conditions of salmon frame were examined, and the characteristics of the extracts were compared with commercial Gomtang. Based on the crude protein, Ex-N and sensory attributes, the extractions were optimized by extracting pretreated-salmon frame in 12 times (v/w) of water for 12 hrs, before filtering with cheese cloth to yield 3 times the volume of the raw material. The concentrations of heavy metals in extracts from salmon frame were below the safety limits suggested by KFDA. The mai or amino acids were glutamic acid and aspartic acid as the free amino acids, and glycine, proline, and glutamic acid as the total amino acids. The calcium and phosphorus contents were 18.0 mg/100 mL and 33.1 mg/100 mL, respectively, and they accounted for 20% and 18% of the recommended daily allowance for mineral intake. The angiotensin I converting enzyme (ACE) inhibitory activity was improved by incubation with Flavourzyme for 4 hrs and its $IC_{50}$ was 2 mg/mL. The results above suggested that the enzymatic hydrolysates from extracts of salmon frame could be used as a basic ingredient for preparing Gomtang-like products.

Biological Activity of Extracts from Chrysanthemum incidicum Linne by Ultrafine Grinding (미세분쇄에 의한 감국(Chrysanthemum incidicum Linne) 추출물의 생리활성)

  • Cho, Young-Je
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.43 no.1
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    • pp.110-117
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    • 2014
  • In this study, the biological activity of water and ethanol extracts from Chrysanthemum incidicum Linne by ultrafine grinding for functional food source are examined. The content of phenolic compounds from Chrysanthemum incidicum Linne were the highest when extracted for 6 hr with 70% ethanol. The extraction yield of water and ethanol extracts were $7.12{\pm}1.61$ mg/g and $7.51{\pm}2.14$ mg/g, respectively. With ultrafine grinding, water and ethanol extracts were $8.63{\pm}1.15$ mg/g and $9.33{\pm}1.35$ mg/g, respectively. In determining anti-oxidative activity of Chrysanthemum incidicum Linne extracts, DPPH of normal grinding extracts was 83.52% and ultrafine grinding was 92.37%. In ABTS radical cation decolorization, normal grinding, fine grinding, and ultrafine grinding extracts were 90% or higher. In antioxidant protection factor (PF), water and ethanol extracts of ultrafine grinding showed relatively high anti-oxidative activities of each 1.82 PF and 2.16 PF, respectively. The TBARS value of ultrafine grinding extracts were lower than normal grinding and fine grinding extracts. The inhibition activity on xanthin oxidase of Chrysanthemum incidicum Linne extracts was 67.53% in ultrafine grinded water extracts and 83.45% in ultrafine grinded ethanol extracts. Inhibition on xanthin oxidase of ethanol extracts showed a higher inhibition effect than water extracts, and ultrafine grinding was higher than normal grinding. In angiotensin converting enzyme inhibition activity, ultrafine grinding water extract was 24% or higher, and ethanol extract was 34% or higher. The elastase inhibition activity of ultrafine grinding extract was 25.56%, which was higher than 20.34% of fine grinding extracts. Water extracts did not show hyaluronidase inhibition activity but ethanol extracts showed 35% of hyaluronidase inhibition activity. The determining expression inhibition of iNOS and COX-2 protein in macrophage by Chrysanthemum incidicum Linne extracts with a Western blot analysis, iNOS and COX-2 protein expression inhibition by Chrysanthemum incidicum Linne ethanol extracts were 40% and 15%, respectively at 100 ${\mu}g/mL$ concentration. The inhibitory patterns of iNOS and COX-2 protein expression was concentration dependent. The result suggests that Chrysanthemum incidicum Linne extracts by ultrafine grinding may be more useful than normal grinding as potential sources due to anti-oxidation, angiotensin converting enzyme and xanthine oxidase inhibition, anti-inflammation effect.

Mycelial Growth Using the Natural Product and Angiotensin Converting Enzyme Inhibition Activity of Pleurotus eryngii (천연물을 이용한 큰느타리 균사배양 및 Angiotensin Converting Enzyme 저해활성)

  • Kang, Tae-Su;Jeong, Heon-Sang;Lee, Myong-Yul;Park, Hee-Joeng;Jho, Taek-Sang;Ji, Seung-Taek;Shin, Myung-Keun
    • The Korean Journal of Mycology
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    • v.31 no.3
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    • pp.175-180
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    • 2003
  • To develop the health/functional food materials, we investigated the cultural condition of mycelial growth on the solid state fermentation using the brown rise, Acanthopanax sp. and Artemisia sp., and also evaluated inhibitory activity of angiotensin converting enzyme (ACE) of hot water extracts from cultured media of Pleurotus eryngii. As the amount of Acanthopanax nnd Artemisia In the cultural media increased, the mycelial growth rate decreased. Especially, addition of Aeantopanax showed marked effect than Artemisia. Moisture contents in three kinds of cultured media were in the range of $10.9{\sim}12.0%$. Crude protein fat and crude fiber content were the highest value in cultured brown rice medium, whereas the mineral contents (Ca, K and P) were higher in the Acanthopanax supplemented (5%) medium than the other media, The extraction yield of the Artemisia supplemented (5%) medium was the highest value of 4.80%, and the pH of hot water extract from cultured brown rice medium showed the lowest value of 6.1. Lightness (L) values in three kinds of extracts from cultured media were in the range of $85.8{\sim}87.1$. Redness (a) value was the highest In the brown rice and Acanthopanax supplemented media, however cultured Artemisia supplemented medium showed the highest value in yellowness (b). In comparison of sugar components analyzed by the thin layer chromatography with three kinds of samples, two spots were detected to be glucose and maltose, respectively. The ACE inhibitory activity of hot water extract from the cultured Acanthopanax supplemented medium showed the highest value at the concentration of $0.2{\sim}1.0\;mg/ml$. These results suggest that the Pleurotus eryngii grew in natural media using brown rice and Acanthopanax can be supplemented to the brown rice medium to enhance its ACE inhibitory activity as health/functional food materials.

Antioxidative Activity of Hot Water Extracts from Medicinal Plants (한약재 열수추출물의 항산화 활성)

  • Ju, Jong-Chan;Shin, Jung-Hye;Lee, Soo-Jung;Cho, Hee-Sook;Sung, Nak-Ju
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.35 no.1
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    • pp.7-14
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    • 2006
  • The antioxidative activity and the related parameters of hot water extracts obtained from 16 medicinal plants were tested. The extraction yield was the highest in Salvia miltiorrhiza Bunge $(36.49\%)$ pH range was $4.00\~5.92$ in all samples. Absorbance at 280 nm was examined to determine aromatic compounds content. The absorbance of $250{\mu}g/mL$ sample was the highest in Prunella vulgaris Linne var. lilacina Nakai (2.872) and below 0.5 in Cirsium maackii, Crataegus pinnatifida Bunge and Zizyphus jujuba Miller. Also, absorbance at 420 nm was high in order of Prunella vulgaris Linne vu. lilacina Nakai (1.312), Zea mays Linne (0.917) and Inula japonica Thunberg (0.725) Total phenolic compounds contents was the highest in Prunella vulgaris Linne var. lilacina Nakai $(5.07{\pm}0.05\;mg/100g)$ and flavonoids contents was 2-fold higher in Salvia miltiorrhiza Bunge $(4.82{\pm}1.16\;mg/100g)$ than the other samples. Electron donating abilities of Zizyphus jujuba Miller, Cornus officinalis Siebold et Zuccarini and Salvia miltiorrhiza Bunge were over $90\%$ at $1000{\mu}g/mL$. Reducing power had similar tendency to electron donating ability while reducing power was significantly lower in samples compared to BHT. Two samples (Prunella vulgaris Linne val. lilacina Nakai and Inula japonica Thunberg) were found to have more than $50\%$ nitrite scavenging effect at $500{\mu}g/mL$ while 8 kinds samples (Zizyphus jujuba Miller, Cornus officinalis Siebold 81 Zuccarini, Chrysanthemum indicum Linne, Prunella vulgaris Linne var. lilacina Nakai, Inula japonica Thunberg, Acanthopanax sessiliflorum Seeman, Salvia miltiorrhiza Bunge and Curcuma longa Linne) showed more than $50\%$ nitrite scavenging effect at $1000{\mu}g/mL$. Prunella vulgaris Linne var. lilacina Nakai showed significantly stronger nitrite scavenging effect than other samples, and its activity was $59.62{\pm}1.573\%$ and $80.58{\pm}0.300\%$ at concentrations of 250 and $1000{\mu}g/mL$, respectively.

A Comparison of the Components and Biological Activities in Raw and Boiled Red Beans (Phaseolus radiatus L.) (생팥과 증자팥의 성분 및 생리활성 비교)

  • Lee, Ryun Kyung;Kim, Mi-Sun;Lee, Ye-Seul;Lee, Man-Hyo;Lee, Jong Hwa;Sohn, Ho-Yong
    • Microbiology and Biotechnology Letters
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    • v.42 no.2
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    • pp.162-169
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    • 2014
  • In the course of study for the development of functional food using red beans (azuki beans, Phaseolus radiatus L.), the ethanol extracts from raw-red bean (RRB) and boiled-red bean (BRB) were prepared, and the components and various biological activities of both were compared. It was observed that the extraction yield, and the total polyphenol content, of the BRB were 1.2 times higher than that of the RRB. However, the contents of total flavonoid, total sugar and reducing sugar in the BRB were 30, 27.9 and 30.8% respectively when compared with those of RRB. In relation to antioxidative activity, both RRB and BRB exhibited moderate DPPH anion, ABTS cation, and nitrite scavenging activities and reducing power, though in all cases RRB demonstrated stronger activities than BRB. The extracts of RRB and BRB did not reveal any antimicrobial activities. In a ${\alpha}$-amylase inhibitory activity assay, RRB was higher than BRB, while BRB showed higher ${\alpha}$-glucosidase inhibitory activity than RRB. A strong and particular activity was observed in an anti-thrombosis activity assay of RRB. The extract of RRB demonstrated strong inhibitions against prothrombin and blood coagulation factors, with moderate thrombin inhibition. However, the extract of BRB did not exhibit any significant anti-thrombosis activity. Our results indicate that RRB has different, but useful biological activities, and loss or elimination of the biologically active substances in RRB occurs during the production of BRB. Therefore, to develop more functional foods from red beans, a study of suitable boiling, heating and drying processes is essential, and the efficient re-use of boiled waste water from the boiling process is necessary. These results could be applied to the further development of functional red bean beverages and sweat red bean pastes.

Physicochemical characteristics and physiological activities of mixture extracts from Liriope platyphylla, Schizandra chinensis, and Panax ginseng C.A. Meyer (맥문동, 오미자 및 인삼 혼합추출물의 이화학적 특성 및 생리활성)

  • Gu, Yul-Ri;Hong, Joo-Heon
    • Food Science and Preservation
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    • v.24 no.3
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    • pp.431-439
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    • 2017
  • This study was conducted to examine the antioxidant activities and physiological activities of mixture extracts (Liriope platyphylla, Schizandra chinensis and Panax ginseng C.A. Meyer) with different extraction mixing ratios (MEC, 2:1:1; ME1, 1:2:1; ME2, 1:1:2; ME3, 1.34:1.33:1.33). The yield of extracts ranged from 25.33 to 33.87%. The total polyphenol and total flavonoid contents of ME1 extracts were 1.01 g/100 g, 0.07 g/100 g, respectively. The total sugar contents of MEC extract was 22.83 g/100 g, respectively. The DPPH and ABTS radical scavenging activities of ME1 extracts at $1,000{\mu}g/mL$ were 26.79% and 21.08%. The superoxide radical scavenging and ferric-reducing antioxidant power of ME1 extracts at $1,000{\mu}g/mL$ were 67.83% and $295.47{\mu}M$, respectively. The functionalities of extracts were investigated with L-132 and RAW264.7 cell lines. The extracts on different mixing ratios did not show the toxicity on L-132 and RAW264.7 cell line in $100-2,500{\mu}g/mL$. The ME1 extract of $1,000{\mu}g/mL$ performed better than other extracts protective effects against oxidative stess in L-132 cells (81.22%) and the ME2 extract at $1,000{\mu}g/mL$ decreased nitric oxide production by $7.48{\mu}M$ which was more potent than other extracts. There results suggest that the ME1 extracts may be a useful functional food material in the food industry.

Manufacturing Fermented Rapeseed Meal Compost using Two Microbial Agents and the Effect of Their Application (유용 미생물 제제 이용 발효 유채박 비료 제조 및 시용 효과)

  • Lee, Ji-Eun;Park, Won;Kim, Kwang-Soo;Lee, Yong-Hwa;Kwon, Da-Eun;Moon, Youn-Ho;Cha, Young-Lok;Kang, Yong-Ku
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.64 no.1
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    • pp.55-62
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    • 2019
  • Rapeseed meal, which is a byproduct of rapeseed oil extraction, improves crop productivity by supplying nutrients to the soil. The present study aimed to manufacture fermented rapeseed meal compost using two effective microbial agents and evaluate their efficiency as fertilizer. To types of fermented rapeseed meal, manufactured using either a bio-carrier or microbial agent, showed no differences in pH, electrical conductivity (EC), and total nitrogen content. However, the contents of $NH_4-N$ and $NO_3-N$ as inorganic nitrogen were increased by 5.6 times and 1.5 times, respectively, after 5 d of fermentation. Rapeseed meal fermented for 5 d was applied to tomato a basal fertilizer and after eight weeks, the plant height increased in all fermented rapeseed treatments compared to that in the chemical fertilizer treatment, and also the quantum yield of photosystem II (PS II) showed the same trend. The total nitrogen content of tomato leaves treated with a microbial fermented rapeseed meal was twice as high as that of that treated with a chemical fertilizer. It was confirmed that the increase in the tomato height was an effect of the rapeseed meal containing inorganic nitrogen, which can easily be absorbed by plants. From these results, it is considered that fermented rapeseed meal manufactured with an effective microbial agent for 5 d showed the highest inorganic nutrient content and greatest growth enhancement in tomato.