• Title/Summary/Keyword: L-cystein

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Purification and Characterization of Polyphenol Oxidase in the Flesh of the Fuji Apple

  • Lim, Jeong-Ho;Jeong, Moon-Cheol;Moon, Kwang-Deog
    • Food Science and Biotechnology
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    • v.15 no.2
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    • pp.177-182
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    • 2006
  • Polyphenol oxidase (PPO) was isolated from the flesh of Fuji apples by DEAE-Cellulose, ammonium sulfate precipitation, phenyl-Sepharose CL-4B, and Sephdex G-100 chromatography. The molecular mass of the purified PPO was estimated to be 40 kDa by SDS polyacrylamide gel electrophoresis. With regard to substrate specificity, maximum activity was achieved with chlorogenic acid as substrate, followed by catechin and catechol whereas, there was no detectable activity with hydroquinic acid, resorcinol, or tyrosine as substrate. The optimum pH and temperature with catechol as substrate were 6.5 and $35^{\circ}C$, respectively. The enzyme was most stable at pH 6.0 and unstable at acidic pH. The enzyme was stable when it was heated to $45^{\circ}C$ but heating at $50^{\circ}C$ for more than 30 min caused 50% loss of activity. Reduced $ZnSO_4$, L-cystein, epigallocatechin-3-o-gallate (EGCG), and gallocatechin gallate (GCG) also inhibited activity.

Phenolic Compounds in Persimmon Fruits and Stabilization of Discoloring Compounds (감의 페놀성 화합물과 변색관련 물질의 안정화)

  • 박용곤;김홍만;강윤한
    • The Korean Journal of Food And Nutrition
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    • v.13 no.2
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    • pp.103-110
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    • 2000
  • This study was conducted to determine changes in phenolic compounds of astrigent persimmons before and after softening process and evaluate discoloring properties of major phenolic compounds. Phenolic compounds in soft persimmons were mainly composed of catechins and chlorogenic acid. Although contents of phenolic compounds were reduced during the softening process, little change in the ratio of catechins to total phenolic compounds was observed. Most of phenolic compounds in damaged astringents persimmons were existed in the high molecular weight fraction and more phenolic compounds were extracted at the temperature higher than room temperature. To evaluate discoloring abilities of phenolic compounds, phenolic compounds were dissolved separately into water or 80% methanol. With presence of various amounts of anti-discoloring agents such as vitamin C, citric acid, and L-cystein, (+)catechin was significantly reduced.

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Effects of Amino Acids on the Inhibition of Polyphenol Oxidase Activity from Perillae Folium (아미노산류가 들깨잎 폴리페놀 옥시다제 활성저해에 미치는 영향)

  • Park, Soo-Sun;Kim, An-Keun;Sohn, Eun-Soo
    • YAKHAK HOEJI
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    • v.40 no.1
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    • pp.65-71
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    • 1996
  • Characterization of Polyphenol oxidase (PPO) in Perillae Folium, particullarly inhibitor studies were investigated. This enzyme was stable at pH 5.0 and the residual activity of PPO at ${\geq}$ ph 5.5 was estimated to be very low. PPO activity was decreased slightly by adding amino acid with catechol as a substrate, particullary PPO activity was inhibited markedly by cystein, histidine, lysine and arginine. In the absorption spectra of the product formed when catechol was oxidized by PPO, with a ${\lambda}_{max}$ at 410nm, the peak shifted toward ${\lambda}_{max}$ 520nm by addition of L-proline. At relatively low concentrations($10^{-3}M$), sulfite and dithiothreithol completely inhibited PPO activity. Inhibition of PPO activity by amino acids and inhibitors increased or decreased depending on the pH used to measure it.

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Statistical Optimization of Medium for Formate-driven Bio-hydrogen Production by the Hyperthermophilic Archaeon, Thermococus onnurineus (초고온성 고세균 Thermococcus onnurineus의 개미산으로부터 바이오수소 생산을 위한 통계적 배지 최적화)

  • Lee, Sung-Mok;Kim, Tae Wan;Lee, Hyun Sook;Lee, Jung-Hyun;Kang, Sung Gyun
    • Ocean and Polar Research
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    • v.39 no.4
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    • pp.269-277
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    • 2017
  • Medium compositions for the hyperthermophilic archaeon, Thermococcus onnurineus NA1 was statistically optimized to enhance formate-driven hydrogen ($H_2$) production by using response surface methodology. From the Plackett-Burman design-based experiment, it was confirmed that among the minor components of medium such as KCl, $MgSO_4$, $NH_4Cl$, Cystein-HCl, trace elements, Fe-EDTA and $CaCl_2$, the trace elements were screened as the only positively effective components with respect to $H_2$ production. Subsequently, the optimal concentrations of the trace elements and the major components of a medium such as NaCl, yeast extract and sodium formate were determined from the five-level central composite design (CCD)-based experiment. The resulting quadratic model predicted the maximum $H_2$ production of 46.6 mmol/L in serum bottle and it was validated experimentally using the optimal medium initially supplemented with 26.70 g/L of NaCl, 9.81 g/L of sodium formate, 3.50 g/L of yeast extract and 4.59 mL/L of trace elements. From the duplicate batch cultivations in the fermentor using the optimized medium, the a maximum $H_2$ production rate up to 71.8 mmol/L/h could be obtained, which was a 65% enhanced value compared with that obtained using the control medium, showing the high efficiency of the optimized medium.

Purification and Characterization of Polyphenol Oxidase from Burdock (Arctium lappa L.) (우엉(Arctium lappa L.) 뿌리 Polyphenol Oxidase의 부분정제 및 특성)

  • Lim, Jeong-Ho;Jeong, Moon-Cheol;Moon, Kwang-Deog
    • Food Science and Preservation
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    • v.12 no.5
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    • pp.489-495
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    • 2005
  • Polyphenol oxidase (PPO) from Burdock (Arctium lappa L.) was purified and characterized. Purification of polyphenol oxidase was achieved by ammonium sulfate precipitation, Phenyl-sepharose CL-4B hydrophobic chromatography and Sephadex G-100 gel filtration chromatography. The molecular mass of the purified PPO was estimated to be 30 kDa by SDS polyacrylamide gel electrophoresis. In a substrate specificity, maximum activity was achieved with chlorogenic acid, followed by catechol and catechin. Whereas, there was low activity with hydroquinic acid, resorcinol or tyrosine. The optimum pH and temperature for enzyme activity were 7.0 and 35$\circC$ with catechol, respectively. The enzyme was most stable at pH 7.0 while unstable at acidic and alkaline pH. The enzyme was stable when heated to 40$\circC$. But heating at 50$\circC$ for more than 30 min caused 50% loss of activity. Ascorbic acid, L-cystein and $Cu^{2+}$ inhibited the activity of pholyphenol oxidase.

Purification and Characterization of Carboxymethyl Cellulase from Stropharia rugosoannulata (독청버섯아재비 균주가 생산하는 Carboxymethyl Cellulase의 정제 및 효소학적 특성)

  • Yoo, Kwan-Hee;Chang, Hyung-Soo
    • The Korean Journal of Mycology
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    • v.30 no.2
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    • pp.113-118
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    • 2002
  • A Carboxymethyl Cellulase (CMCase) has been isolated and purified from the edible mushroom, Stropharia rugosoannulata. The molecular weight of CMCase was estimated to be 54 kDa by SDS polyacryl amide gel electrophoresis. The maximum activity of the purified CMCase was observed at pH 4.0 and $40^{\circ}C$, and stable for pH 3.0 to 11.0 to maintain 40% activity. The CMCase activity was activated by $AgNO_{3},\;MgSO_{4},\;and\;KCl$. However, its activity was inhibited by 1,10-phenanthroline, KCN and L-cysteine. Also, the enzyme activity was decreased by the addition of EDTA, suggesting that the purified CMCase is metalloenzyme.

Utilization of Ascidian, Halocynthia roretzi -4. Browning of Ascidian meat, Halocynthia roretzi and Its Prevention- (우렁쉥이 이용에 관한 연구 -4. 우렁쉥이 육의 갈변 및 그 방지-)

  • LEE Kang-Ho;CHO Ho-Sung;KIM Dong-Soo;HONG Byeong-Il;PARK Cheon-Soo;KIM Min-Gi
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.26 no.3
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    • pp.214-220
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    • 1993
  • Browning of ascidian, Halocynthia roretzi, meat occurres very rapidly when skinned off or cut during processing and it resulted the quality loss of fresh frozen, dehydrated or fermented products. In this study, the causes of color development and prevention of browning were experimented. The browning of ascidian meat may be occurred enzymatically by a tyrosinase contained in meat and viscera which acted specifically on L-tyrosine as a substrate rather than on catechol. Activity of the enzyme in viscera was three times higher than in meat. The optimum pH and temperature on the tyrosinase activity of crude enzyme obtained from ascidian was 6.0 and $30{\sim}35^{\circ}C$, respectively. The enzyme was inactivated heating at $80^{\circ}C$ for 3 minutes or $90{\sim}100^{\circ}C$ for 1 minute and it was inhibited by $0.1{\sim}0.5mM$ solutions at ascorbic acid, sodium hydrogen sulfite, cystein, citric acid, cyanide but only sodium hydrogen sulfite treatment was effective to retard such a high content of enzyme as in case of viscera. In practical use for processing of ascidian meat browning was retarded by dipping the viscera removed ascidian meat in 0.2M citric acid for 5 minutes or $0.2\%$ sodium hydrogen sulfite solution for 1 minute resulting in sulfur dioxide residue less than 100 ppm.

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Studies of separation and quantitation for selenium species in food (식품중의 셀레늄 화학종의 분리 및 정량연구)

  • Jang, Hee-Young;Min, Hyungsik;Lee, Jonghae;Pak, Yong-Nam
    • Analytical Science and Technology
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    • v.26 no.3
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    • pp.182-189
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    • 2013
  • The purpose of this research is to separate and quantitate selenium species in some food samples with HPLC-ICP-MS. Cation exchange chromatography showed efficient separation only for inorganic Se species while reversed phase ion pair chromatography showed good separation for both inorganic and organic Se species. $C_8$ column ($Symmetryshield^{TM}\;RP_8$, 3.5 ${\mu}m$, $4.6{\times}150$ mm) was used with optimum condition of 5% methanol mobile phase, 0.05% of nonafluorovaleric acid ion pairing reagent. Five standard Se species of Se(IV), Se(VI), SeCys(selenocystein), SeMet(selenomethionine) and Se-M-C(seleno methyl cystein) were separated successfully under the optimum condition (mobile phase; 5% methanol, ion-pairing reagent; 0.05% nonafluorovaleric acid, flow rate; 0.9 mL $min^{-1}$). To extract Se species, microwave assisted and enzyme-assisted extraction methods were studied. In enzyme-assisted extraction method, protease I for garlic, protease I plus trypsin for pork and mackerel, and protease XIV for tuna showed the best extraction efficiency. With the optimum condition for each sample, it was found that mostly inorganic Se, SeCys and SeMet are present in the sample studied ranging from few ${\mu}g$ $g^{-1}$ to few tens of ${\mu}g$ $g^{-1}$.

Expression of Functionally Human Interleukine-18 by Tobacco Plant Cell

  • Im, Yeong-Lee;Gwon, Tae-Ho;Park, Seung-Mun;Kim, Dae-Hyeok;Jang, Yong-Seok;Yang, Mun-Sik
    • 한국생물공학회:학술대회논문집
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    • 2001.11a
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    • pp.193-196
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    • 2001
  • IL-18. formerly known as IGIF(interferon -gamma inducing factor), is structurally IL-l related but functionally IL-12 related pro-inflammatory cytokine. The human IL -18(hIL-lS), like IL-$1{\beta}$, is synthesized as a biologically inactive precursor of 24kDa lacking a signal peptide, and then cleaved into an active mature form by cystein protease IL-$1{\beta}$ converting enzyme (ICE: caspase- 1), We tested if the mature hIL -18 can be expressed and secreted into culture medium by transforming the forming gene construct consisting of a mature hIL-18 gene fused to signal peptide of rice amylase lA. Secondly, we were tested if the pro- IL-18 could be processed into a biologically active form by caspase-l like protease in plant. Cell suspension culture was established from the leaf-derived calli of transgenic tobacco plant. Southern and Northern blot analysis indicated the expression of both pro-hIL-18 and mature hIL-18 plant cells. Western blot analysis introduced the protein products of pro- hIL -18 and mhIL -18 were observed in transigenic cell lines. In addition, the molecular size of recombinant pro-hILl-18 and mhIL-18 were estimated to be 24kDa and 18kDa, respectively. ELISA revealed that the amount of pro- hIL -18 was 1.3ug per gram of fresh weight calli. Moreover, the presence of mhIL-18 was detected in the culture medium and it appeared to be 25ug/L.

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Determination of S-Allyl-L-cystein, Diallyl Disulfide, and Total Amino Acids of Black Garlic after Spontaneous Short-term Fermentation (자가숙성발효 후 흑마늘의 S-Allyl-L-cystein, Diallyl Disulfide 및 Total Amino Acids 분석)

  • Kim, Mun-Su;Kim, Min-Ju;Bang, Woo-Suk;Kim, Keun-Sung;Park, Sung-Soo
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.41 no.5
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    • pp.661-665
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    • 2012
  • Garlic (Allium sativum L.) is one of the oldest cultivated plants and has been used throughout the world as a food supplement and a folk medicine for thousands of years. Raw garlic has been processed into a variety of commercial garlic products for consumer convenience. The latest new processing technology, 'spontaneous short-term fermentation', has been developed to process raw garlic into black garlic. The physiologically active effects of garlic have been attributed to its organosulfur compounds. In this study, the proximate compositions and the total amino acid content of raw Namhae garlic and black garlic were determined. The two major organosulfur compounds of garlic, $S$-allyl-L-cysteine (SAC), and diallyl-disulfide (DADS), were also analyzed using RP-HPLC. The proximate compositions were not different between raw and black garlic. The amount of 13 amino acids was greater in black garlic than in raw garlic among a total of 17 amino acids considered. The black garlic had 2-fold higher levels of SAC and 30-fold higher levels of DADS than the raw garlic. Therefore, it is suggested that consuming black garlic produced by spontaneous short-term fermentation is more effective than consuming raw garlic, in order for consumers to take more physiologically active organosulfur compounds (SAC and DADS), which are the compounds that are good for consumer health.