• Title/Summary/Keyword: enzymatic color reaction

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Effective Microwell Plate-Based Screening Method for Microbes Producing Cellulase and Xylanase and Its Application

  • Kim, Jennifer Jooyoun;Kwon, Young-Kyung;Kim, Ji Hyung;Heo, Soo-Jin;Lee, Youngdeuk;Lee, Su-Jin;Shim, Won-Bo;Jung, Won-Kyo;Hyun, Jung-Ho;Kwon, Kae Kyoung;Kang, Do-Hyung;Oh, Chulhong
    • Journal of Microbiology and Biotechnology
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    • v.24 no.11
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    • pp.1559-1565
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    • 2014
  • Cellulase and xylanase are main hydrolysis enzymes for the degradation of cellulosic and hemicellulosic biomass, respectively. In this study, our aim was to develop and test the efficacy of a rapid, high-throughput method to screen hydrolytic-enzyme-producing microbes. To accomplish this, we modified the 3,5-dinitrosalicylic acid (DNS) method for microwell plate-based screening. Targeted microbial samples were initially cultured on agar plates with both cellulose and xylan as substrates. Then, isolated colonies were subcultured in broth media containing yeast extract and either cellulose or xylan. The supernatants of the culture broth were tested with our modified DNS screening method in a 96-microwell plate, with a $200{\mu}l$ total reaction volume. In addition, the stability and reliability of glucose and xylose standards, which were used to determine the enzymatic activity, were studied at $100^{\circ}C$ for different time intervals in a dry oven. It was concluded that the minimum incubation time required for stable color development of the standard solution is 20 min. With this technique, we successfully screened 21 and 31 cellulase- and xylanase-producing strains, respectively, in a single experimental trial. Among the identified strains, 19 showed both cellulose and xylan hydrolyzing activities. These microbes can be applied to bioethanol production from cellulosic and hemicellulosic biomass.

Non Thermal Process and Quality Changes of Foxtail Millet Yakju by Micro Filtration (미세여과에 의한 비 가열살균 좁쌀약주의 제조 및 저장 중 품질변화)

  • Kang, Young-Joo;Oh, Young-Ju;Koh, Jeong-Sam
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.34 no.2
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    • pp.277-284
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    • 2005
  • Micro-filtration (MF) or ultra-filtration (UF) system with hollow-fiber cartridge was introduced in order to improve the Quality level of commercial foxtail millet Yakju, which has an off-flavour and/or undesired colour after the thermal treatment. The filtration effects of cartridges such as MF (0.65, 0.45, 0.2, 0.1 $\mu$m) and UF (500 K dalton) were investigated. The physicochemical and sensory characteristics of the Yakju were then evaluated during the 6 months storage at room temperature. The exclusion ability of microorganism in samples was confirmed in all cartridges, but 0.45 pm MF-cartridge was suitable in the Yakju manufacture due to its superior filtration rate and efficiency. Changes in reducing sugar and colour difference of foxtail millet Yakju untreated or treated by heat ($65^{\circ}C$${\times}$10 min) were observed during the storage; after 6 months the L-value of thermal-treatment sample was decreased and its b-value, however, significantly increased so that its color became dark, in comparison to non-thermal treatment sample. This decrease of reducing sugar is assumed that color change is associated with non-enzymatic browning reaction. Sensory Quality of foxtail millet Yakju produced by non-thermal treatment was better than that of thermal treatment.

Change in Quality of Mixed Juice of Fruits and Vegetables by Aseptic Treatment and Packing with Nitrogen Gas during Storage (제균처리와 질소가스포장에 따른 혼합과채주스의 저장 중 품질 변화)

  • Kim, Su-Yeun;Yoon, Young-Bean;Choi, Eon-Ho
    • Korean Journal of Food Science and Technology
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    • v.32 no.6
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    • pp.1271-1277
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    • 2000
  • The extracts from tomatoes, apples, carrots, mallows, watercreses+pine needles, Angelica keiskei Koiz, jujubes and lemons were selected and mixed at the ratio of 3 : 3 : 3 : 1/2 : 1/2 : 1/2 : 1/2 : 1/5 on the basis of sensory evaluation. The mixed extracts were divided into three lots and treated as follows. The first lot was heated for 15 sec at $96^{\circ}C$ without centrifugation and filtration, the second filtered through a ultramembrane filter. For the third one, the ultrafiltrate combined with autoclaved retentates on the membrane filter and the previous centrifugal precipitation. The mixed juices were stored in the glass bottles with atmosphere or in film package with nitrogen gas. And then they were stored at $4^{\circ}$ and $20^{\circ}C$. During storage the treated juices showed $pH\;4.07{\sim}4.10$, titratable acidity $66.35{\sim}84.08$, soluble solid $7{\sim}9^{\circ}Brix$, reducing sugar $5.42{\sim}6.97%$, glucose $1.96{\sim}2.30%$ and fructose $3.46{\sim}4.14%$. The ultrafiltered juices showed yellow color, different from orange color of other treatment lots. Peroxidase activity and microbial population were inhibited by thermal treatment and ultrafiltration. The browning effects of the mixed juice of fruits and vegetables during storage were caused by ascorbic acid oxidation and non-enzymatic browning reaction, which could be reduced by change and packing with nitrogen gas instead of atmosphere inside bottles.

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Conditions for Rapid Processing of Modified Fish Sauce using Enzymatic Hydrolysis and Improvement of Product Quality 1. Fish Sauce from Mackerel Waste and Its Quality (효소분해법에 의한 개량어장유의 속성제조 및 품질에 관한 연구 1. 고등어 폐기물을 이용한 어장유의 속성제조 및 품질)

  • HAN Bong-Ho;BAE Tae-Jin;CHO Hyun-Duk;KIM Jong-Chul;KIM Byeong-Sam;CHOI Soo-Il
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.23 no.2
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    • pp.109-124
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    • 1990
  • A rapid processing method for fish sauce of high quality stability and favorable flavor was investigated using mackerel waste as starting material. The chopped waste was homogenized with water and hydrolyzed by commercial proteolytic enzymes such as Complex enzyme-2000($2.18\cdot10^4$ U/g solid, Pacific Chem. Co.) and Alcalase ($1.94\cdot10^4$ U/g solid, Novo) in a cylindrical vessel with 4 baffles and 6-bladed turbine impeller. Optimal pH and temperature for the hydrolysis with Complex enzyme-2000 were 8.0 and $50^{\circ}C$, and those with Alcalase were 9.0 and $55^{\circ}C$. In both cases, the reasonabe amount of added water and enzyme concentration based on the waste weight were $40\%,\;3\%$ and hydrolyzing time was 100 min. Thermal treatment of the hydrolysate with $6\%$ of invert sugar for 2 hours at $90^{\circ}C$ was adequated to inactivation of the enzymes and pasteurization of the hydrolysate. Flavor, taste and color of the hydrolysate were improved during the thermal treatment in which the browning reaction products might participate and result in antioxidative and bactericidal effects. Combined use of $0.005\%$ of Caryophylli flos with $6\%$ of invert sugar was also effective for the improvement of taste. Yield of the fish sauce based on the total nitrogen of the raw waste was $93.7\~94.9\%$, and $87.6\~87.9\%$ of the total nitrogen in the fish sauce was in the from of amino nitrogen. The pH, salinity and histamine content of the fish sauce prepared with $15\%$ of table salt were $6.1\~6.2$, $14.0\~14.5\%$ and less than $10mg\%$, respectively. The fish sauce was stable on bacterial growth during the storage of 60 days at $26\pm3^{\circ}C$ and the quality was also maintained.

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Conditions for Rapid Processing of Modified Fish Sauce using Enzymatic Hydrolysis and Improvement of Product Quality 2. Fish Sauce from Sardine Waste and Its Quality (효소분해법에 의한 개량어장유의 속성제조 및 품질에 관한 연구 2. 정어리 폐기물을 이용한 어장유의 속성제조 및 품질)

  • BAE Tae-Jin;HAN Bong-Ho;CHO Hyun-Duk;KIM Jong-Chul;KIM Byeong-Sam;CHOI Soo-Il
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.23 no.2
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    • pp.125-136
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    • 1990
  • To develope a rapid processing method for fish sauce, processing conditions of fish sauce from sardine waste was investigated. The chopped waste was homogenized and hydrolyzed by commercial proteolytic enzymes such as Complex enzyme-2000($2.18\cdot10^4$ U/g solid) and Alcalase($1.94\cdot10^4$ U/g solid) in a cylindrical vessel with 4 baffles and 6-bladed turbine impeller. Optimal temperature for the case of hydrolysis with Complex enzyme-2000 was 50 and that with Alcalase was $55^{\circ}C$. In both cases, the reasonable pH, amount of water for homo-genization, enzyme concentration and hydrolyzing time were 8.0, $40\%$ (W/W), $3\%$ and 100 min, respectively. Heating of the filtrated hydrolysate for 2 hours at $90^{\circ}C$ with $6\%$ of invert sugar was suitable for pasteurization of the hydrolysate and inactivation of enzymes. Flavor, taste and color of the hydrolysate was improved during the thermal treatment in which the browning reaction products might participate and result in antioxidative and bactericidal effects. Combined use of $0.005\%$ of Caryophylli flos with invert sugar was also effective for the improvement of taste. Yield of the fish sauce based on the total nitrogen in the raw sardine waste was $91.2\~92.3\%$ and $87.2\~87.8\%$ of the total nitrogen in the fish sauce was in the form of amino nitrogen. The pH, salinity and histamine content of the fish sauce prepared with $15\%$ of table salt were $6.1\~6.2$, $14.2\~14.4\%$ and less than $10mg\%$, respectively. The fish sauce was stable during the storage of 60 days at $26\pm3^{\circ}C$ on bacterial growth and its quality was also maintained.

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Immunostimulatory activity and structural characteristics of neutral polysaccharides isolated from ginseng leaves fermented by Cordyceps sinensis (동충하초균으로 발효한 인삼잎에서 분리한 중성다당의 면역활성 및 구조적 특성)

  • Cha, Ha Young;Park, Hye-Ryung;Shin, Kwang-Soon
    • Korean Journal of Food Science and Technology
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    • v.53 no.5
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    • pp.570-577
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    • 2021
  • This study elucidated the biological activities and structural characteristics of polysaccharides isolated from ginseng leaves fermented using Cordyceps sinensis (GLF). GLF comprised at least 18 glycosyl linkages, including 4-linked glucose residues (24.0%). To characterize the neutral polysaccharides in GLF, it was further fractionated by anion exchange chromatography, and the unabsorbed fraction (GLF1) was isolated. Peritoneal macrophages stimulated with GLF1 produced various cytokines in a dose-dependent manner. The properties and activities of the four subfractions (PHI, PHIA1-PHIA3) obtained after sequential enzymatic digestion were examined. PHI and PHIA3 primarily comprised glucose, whereas PHI exhibited an iodine-color reaction. Furthermore, the PHIA1-3 fractions indicated that cytokine production was completely inhibited. These results suggest at the immune activities of GLF1 may be due to the α-(1→4)-glucan branched at the C(O)6 position, which was produced by C. sinensis.

Synthesis and Functional Properties of Plastein from the Enzymatic Hydrolysates of Filefish Protein 2. General Properties and IR Spectrum of Plasteins (말쥐치육 단백질의 효소적 가수분해물을 이용한 Plastein의 합성 및 그 물성 2. Plastein의 일반적 성상과 IR Spectrum)

  • KIM Se-Kwon;LEE Eung-Ho
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.20 no.5
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    • pp.431-440
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    • 1987
  • In order to develop a new type of food source for the effective utilization of fish protein, plastein reaction was applied to improve the functional properties of filefish protein. Plasteins were synthesized from a peptic filefish protein hydrolysate by papain, pepsin, $\alpha-chymotrypsin$ and protease(from Streptomyces griceus) under the optimum conditions of previous paper). Also, L-glutamic acid diethylester and L-leucine ethylester were incorporated into plastein during the plastein reaction by papain. And, General composition, yield, molecular weight, amino acid composition, color and IR spectrum of plasteins were measured. The protein, ash and lipid content of the plasteins were $72\~78\%,\;7.4\~11.8\%\;and\;0.3\~0.9\%$ respectively. The yield of plasteins were papain $55.0\%,\;pepsin\;47.6\%,\;\alpha-chymotrypsin\;38.3\%,\;protease\;23.6\%$, glutamic acid-incorporated plastein (Glu-Plastein) $35.0\%$, and leucine-incorporated plastein (Leu-plastein) $45.7\%$. The glutamic acid and leucine content in Glu-plastein and Leu-plastein were $38.7\%,\;41,7\%$, respectively, while the contents in the peptic filefish protein hydrolysate were $16.01\%\;and\;8.16\%$, respectively. The amino acid compositions were similar to that of the original filefish muscle protein. The major molecular weights of the peptic hydrolysate estimated by gel filteration were 2,000 and 310, and those of plasteihs were 21,000 and 4,900 for papain, 24,000 for pepsin, 18,500 for $\alpha-chymotrypsin$ 6,700 for protease, 24,000 for Glu-plastein and 17,000 for Leu-plastein. The structural changes in freeze-dried filefish meat, the FPC and hydrolysate were not observed on the IR spectrum. But plasteins showed amide I band in $1,600\~l,700cm^{-1}$ range and resulted in a strong band in $800\~850\;cm^{-1},\;700\~750\;cm^{-1}\;and\;650\~700\;cm^{-1}$. The amide I band of Glu-plastein was wider than those of other plasteins and had also a small band at $1,440\;cm^{-1}$.

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