• Title/Summary/Keyword: Enzymatic method

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Enzymatic Synthesis of Resveratrol α-Glucoside by Amylosucrase of Deinococcus geothermalis

  • Moon, Keumok;Lee, Seola;Park, Hyunsu;Cha, Jaeho
    • Journal of Microbiology and Biotechnology
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    • v.31 no.12
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    • pp.1692-1700
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    • 2021
  • Glycosylation of resveratrol was carried out by using the amylosucrase of Deinococcus geothermalis, and the glycosylated products were tested for their solubility, chemical stability, and biological activities. We synthesized and identified these two major glycosylated products as resveratrol-4'-O-α-glucoside and resveratrol-3-O-α-glucoside by nuclear magnetic resonance analysis with a ratio of 5:1. The water solubilities of the two resveratrol-α-glucoside isomers (α-piceid isomers) were approximately 3.6 and 13.5 times higher than that of β-piceid and resveratrol, respectively, and they were also highly stable in buffered solutions. The antioxidant activity of the α-piceid isomers, examined by radical scavenging capability, showed it to be initially lower than that of resveratrol, but as time passed, the α-piceid isomers' activity reached a level similar to that of resveratrol. The α-piceid isomers also showed better inhibitory activity against tyrosinase and melanin synthesis in B16F10 melanoma cells than β-piceid. The cellular uptake of the α-piceid isomers, which was assessed by ultra-performance liquid chromatography (UPLC) analysis of the cell-free extracts of B16F10 melanoma cells, demonstrated that the glycosylated form of resveratrol was gradually converted to resveratrol inside the cells. These results indicate that the enzymatic glycosylation of resveratrol could be a useful method for enhancing the bioavailability of resveratrol.

Effect of Gamma Ray Irradiation on the Pretreatment and Enzymatic Hydrolysis of Senna tora Stalk (감마선 조사 처리에 의한 결명자 줄기의 전처리와 효소가수분해 효과)

  • Kim, Jo Eun;Gong, Sung Ho;Jung, Jin Tae;Lee, Ok Ran;Lee, Jae Won
    • Korean Journal of Medicinal Crop Science
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    • v.26 no.2
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    • pp.127-133
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    • 2018
  • Background: The demand of recycling renewable agricultural by-products is increasing. Radiation breeding is a method used to improve saccharification efficiency. Thus, we investigated the effect of gamma ray irradiation on the pretreatment and enzymatic hydrolysis of the stalks of Senna tora, an important medicinal plants. Methods and Results: S. tora seeds were irradiated with gamma ray at doses of 100, 200, 300, and 400 Gy. In the pretreated biomass, glucan and lignin content were higher in the M1 ($1^{st}$ generations of irradiation) S. tora stalks than in the M2 ($2^{nd}$ generations of irradiation) stalks, this can be explained by the higher degradation rate in M1. After oxalic acid pretreatment, the concentration of total phenolic compounds (TPCs) in the hydrolysate increased in the gamma ray treated seeds. The highest relative increase rate in crystallinity in the pretreated biomass was observed in M1-400 Gy and M2-100 Gy. The cellulose conversion rate was higher in M1 than in M2, except for 200 Gy. Conclusions: Gamma ray irradiation at an appropriate dose can be used to improve the efficiency of pretreatment and enzymatic hydrolysis, thereby increasing biomass availability.

Effect of High Speed Drying on Antioxidant Properties of Enzymatic Digests from Citrus By-products and Their Protective Effect on DNA Damage Induced by $H_2O_2$

  • Senevirathne, Mahinda;Kim, Soo-Hyun;Um, Byung-Hun;Lee, Jung-Suk;Ha, Jin-Hwan;Lee, Won-Woo;Jeon, You-Jin
    • Food Science and Biotechnology
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    • v.18 no.3
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    • pp.672-681
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    • 2009
  • The aim of this study was to evaluate the antioxidant activities of enzymatic digests from citrus by-products (CBPs) prepared by high speed drying (HSD). HSD needs a short time (60 min) for drying and can be used in a commercial scale. Enzymatic digests were prepared from the CBPs using 6 enzymes such as aminoglucosidase (AMG), celluclast, pectinex, termamyl, ultraflo, and viscozyme. Antioxidant activities of AMG digest from CBPs were evaluated by different in vitro models such as 1,1-diphenyl-2-picrylhydrazyl (DPPH), hydroxyl, alkyl, $H_2O_2$ scavenging, metal chelating, lipid peroxidation, and comet assays, and exhibited strong activities. The antioxidant compounds were detected by an high performance liquid chromatography (HPLC) coupled on-line to an 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid ($ABTS^+$) scavenging detection system, and found that hesperidin was the key compound. Therefore, the results obtained in this study suggest that HSD is an effective method to transform wet CBPs into dried form, and CBPs are potential source of natural antioxidant.

High-Solid Enzymatic Hydrolysis and Fermentation of Solka Floc into Ethanol

  • Um, Byung-Hwan;Hanley, Thomas R.
    • Journal of Microbiology and Biotechnology
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    • v.18 no.7
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    • pp.1257-1265
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    • 2008
  • To lower the cost of ethanol distillation of fermentation broths, a high initial glucose concentration is desired. However, an increase in the substrate concentration typically reduces the ethanol yield because of insufficient mass and heat transfer. In addition, different operating temperatures are required to optimize the enzymatic hydrolysis (50$^{\circ}C$) and fermentation (30$^{\circ}C$). Thus, to overcome these incompatible temperatures, saccharification followed by fermentation (SFF) was employed with relatively high solid concentrations (10% to 20%) using a portion loading method. In this study, glucose and ethanol were produced from Solka Floc, which was first digested by enzymes at 50$^{\circ}C$ for 48 h, followed by fermentation. In this process, commercial enzymes were used in combination with a recombinant strain of Zymomonas mobilis (39679:pZB4L). The effects of the substrate concentration (10% to 20%, w/v) and reactor configuration were also investigated. In the first step, the enzyme reaction was achieved using 20 FPU/g cellulose at 50$^{\circ}C$ for 96 h. The fermentation was then performed at 30$^{\circ}C$ for 96 h. The enzymatic digestibility was 50.7%, 38.4%, and 29.4% after 96 h with a baffled Rushton impeller and initial solid concentration of 10%, 15%, and 20% (w/v), respectively, which was significantly higher than that obtained with a baffled marine impeller. The highest ethanol yield of 83.6%, 73.4%, and 21.8%, based on the theoretical amount of glucose, was obtained with a substrate concentration of 10%, 15%, and 20%, respectively, which also corresponded to 80.5%, 68.6%, and 19.1%, based on the theoretical amount of the cell biomass and soluble glucose present after 48 h of SFF.

Physicochemical Characteristics of Silk Fibroin Degummed by Protease in Bacillus licheniformis II. Effect of Heat Treatment onto Degummed Silk Fiber (Bacillus licheniformis 단백질 분해 효소에 의한 정련 견사의 특성 II. 정련 견사의 열처리 영향)

  • 김영대;남중희
    • Journal of Sericultural and Entomological Science
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    • v.35 no.1
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    • pp.52-59
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    • 1993
  • Raw silk degumming, by which the sericin and other marterials are eliminated from fibroin, is very essential process to produce silk fabrics. Alkali chemicals and enzymes have been used for the silk-degumming process. In this paper, the effect of heat treatment was investigated on silk fibers degummed by two different methods, soap and enzymatic degumming method. The difference between these two degumming methods was analyzed on the basis of results of mechanical testing, thermal analysis and intrated spectroscopy. The tenacity and the elongation of silk fiber are decreased by the heat-treatment in wet state. This tendency is observed in both cases of two degumming methods. The peak temperature in DSC analysis, which is attributed to thermal decomposition of silk fiber, was shifted to higher value with the heat-treatment temperature for the soap degummed silk fiber, however, it was not for the enzymatic degummed one. The IR crystallinity of soap digummed silk fiber is increased with the heat-treatment temperature, while that of enzymatic degummed fiber is not.

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Microbial Extracellular Enzyme Detection on Agar Plates by Means of Fluorogenic Methylumbelliferyl-Substrates (Methylumbelliferyl 형광기질을 이용한 평판배지상의 미생물 체외 세포효소측정방법)

  • ;Hoppe, H.-G.
    • Korean Journal of Microbiology
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    • v.28 no.3
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    • pp.229-235
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    • 1990
  • A rapid and sensitive method to detect the extracellular enzymatic activity of bacteria colonies grown on agar plates is described. Selective agar media supplemented with protein, starch, chitin, Tween-80, etc. are conventionally used to detect biochemical properties of bacteria. It has been experimentally demonstrated with bacteria pure cultures that fluorogenic Methylumbelliferyl (MUF)-substrates are excellent substrate analogues for normally occurring polymers. Based on MUF-substrate hydrolysis the new method provides reliable qualitative estimates of extracellular enzymatic properties of bacteria within minutes using pure cultures as well as agar p;ates prepared for colony counts. Extracellular enzyme activities of heterotrophic bacteria from freshwater ecosystems and marine sediment using this method are discussed.

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Production of L-Tryptophan by Enzymatic Processes (효소공정에 의한 트립토판 생산)

  • 이인영;안경섭;김의환;이선복
    • Microbiology and Biotechnology Letters
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    • v.20 no.1
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    • pp.73-78
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    • 1992
  • - Enzymatic synthesis of L-tryptophan(Trp) using E. coli tryptophanase has been investigated. In order to reduce the substrate inhibition by indole and to increase the product yield of L-tryptophan three different approaches have been made in this work. First, indole was intermittently fed to the reaction mixture in order to control the indole concentration at lower level. When 15 mM of indole was used as a total amount of substrate, conversion yield of 80% has been obtained with intermittent feeding while only 20% of indole was converted into L-tryptophan by conventional batch operation, The second method employed in this work was the use of cyclohexane-phosphate buffer organic two-phase system. In this system, indole was mainly partitioned into the organic-solvent phase and therefore substrate inhibition was expected to be reduced. L-Tryptophan production in organic two-phase system was, however, unexpectedly lower than that obtained in aqueous buffer solution. As a third method cyclodextrins have been added to the aqueous reaction mixture. It was found that the addition of $\beta$-cyclodextrin enhanced the tryptophan synthesis noticeably while $\alpha$-cycfodextrin showed little effect on tryptophan production.

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Quantitative Determination of Total Bile Acids from Bezoar and Bezoar-containing Liquid Preparation by Enzymatic Technique (효소반응법을 이용한 우황 및 우황함유 액상 제제 중 총담즙산의 정량)

  • Ha, In-Sik;Kim, Seung-Hwan;Cha, Bong-Jin;Kwon, Jong-Won;Yang, Joong-Ik;Min, Shin-Hong
    • Journal of Pharmaceutical Investigation
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    • v.21 no.2
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    • pp.67-71
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    • 1991
  • A simple and sensitive method was developed for the quantification of free and conjugated bile acids in bezoar without prior hydrolysis. $3{\alpha}-Hydroxy$ bile acids are first oxidized to 3-keto bile acids in the reaction catalyzed by $3{\alpha}-hydroxysteroid$ $dehydrogenase(3{\alpha}-HSD)$. During this oxidative reaction, an equimolar quantity of nicotinamide adenine dinucleotide(NAD) is reduced to NADH and subsequently oxidized to NAD with concomitant reduction of nitrotetrazolium blue(NTB) to diformazan by the catalytic action of diaphorase. The diformazan has an absorbance maximum at 540 nm. The intensity of the color produced is directly proportional to bile acids concentration in the bezoar extracts. The optimum conditions for the enzymatic reaction such as effects of reaction time, reaction temperature and pH, and stability were investigated. Calibration plots for the sodium chelate observed to be linear and intra-, inter-assay analytical recovery of bile acids averaged $97.65{\pm}3.4%(S.D.)$. Therefore, it is considered that the quality control of total bile acids from bezoar or bezoar-containing liquid preparation using this simple and sensitive assay system will be acceptable. Also current bezoars and bezoar-containing liauid preparations were examined their total bile acids from this method.

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효소처리에 의한 제지적성 개선

  • 김형진;조병묵
    • Proceedings of the Korea Technical Association of the Pulp and Paper Industry Conference
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    • 2000.04a
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    • pp.39-46
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    • 2000
  • In pulp and papermaking process, enzymatic treatment of pulp fibres has been a topic of increasing interest in last decade. Lots of patents, papers and research reports were published on the application of enzymes in the fields of enzymatic bleaching, deinking, slime control, pitch control, waste water treatment and fibre modification. Cellulase and hemicellulase are the principal enzymes used for the modification of fibre property. This study was carried out for determinating the behaviors of enzyme to pulp fibres. A commercial enzyme, Denimax BT which is consisted with cellulase and hemicellulase, was treated to the kraft pulp produced from domestic hardwood mixtures. Results were mainly concentrated on the behaviors of freeness, drainability and fines content of fibres, and physical properties of paper with enzyme treatment. The freeness levels and dewatering ability were developed, and the fines contents were decreased. The creation of fines were controlled by the method of pre-enzyme treatment prior to fibre beating. The mechanical strength of paper, like tensile, burst, tear strength and folding endurance, were remarkably improved by the pre-enzyme treatment.

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Deinking of Computer Printed Out and White Ledger (백상지 고지의 탈묵)

  • 한신호
    • Journal of Korea Technical Association of The Pulp and Paper Industry
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    • v.32 no.1
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    • pp.48-56
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    • 2000
  • The objective of this study was to investigate the enzymatic deinking technology. Office recycled papers such as computer printed out(CPO) and white ledger(WL) were used in the evaluation of four different deinking conditions ; alkaline deinking , neutral deinking and enzymatic deinking at alkaline and neutral pH. Filltering pads were prepared after flotation deinking to measure brighteness gain and ink removal efficiency. Three different surfactants were also used in the evaluation of deinking efficiency including surface chemical properties by measuring foam height and stability, surface tension and cloud point of views. The results indicated that the neutral deinking method with enzyme and surfactant addition showed highest deinking efficiency of office recycled papers.

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