• Title/Summary/Keyword: enzymatic digestion

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Comparative Study on Development of Mouse Embryos in Three Commercial Media and Hatching Rates of Mouse Embryos with/without Pronase (3개의 배양액내에서 생쥐배아의 발달과 Pronase로 처리한 생쥐배아 부화율의 비교 연구)

  • Lee, Jeong-Heon;Go, Hee-Jeong;Chae, Geu-Jeong;Lee, Ki-Suk;Kim, Jong-Duk
    • Clinical and Experimental Reproductive Medicine
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    • v.28 no.3
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    • pp.235-245
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    • 2001
  • Objectives: The purpose of this present study was to compare mouse embryo development in 3 commercial media and hatching competence of mouse embryo with or without enzymatic treatment. Methods: Collected 375 mouse embryos were divided into three groups, and then cultured in IVF-20 (G2), Medicult IVF (M3), P-1 (blastocyst M), respectively. Three day mouse morulae were cultured in G2 media treated with pronase. The results were analyzed using Chi-square test, and considered statistically significant when p<0.01. Results: The developmental rate of 2 cell mouse embryo after 72 hours was highest in IVF-20 (G2) among conventional 3 media. The hatching rate of mouse morulae was low when clultured in G2 media without pronase during 48 hours. However, it was higher when cultured in media treated with $1{\mu}g/ml$, $2.5{\mu}g/ml$, $5{\mu}g/ml$ pronase, respectively. Conclusions: Using good media and digestion of zona pellucida with enzymatic treatment improve development and hatching rate of embryo. Therefore, implantation and pregnancy rate could be improved.

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Microbial and Enzymatic Properties Related to Brewing of Traditional Ewhaju (전통 이화주의 양조와 관련된 미생물 및 효소적 특성)

  • 김정옥;김종군
    • Korean journal of food and cookery science
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    • v.9 no.4
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    • pp.266-271
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    • 1993
  • In order to investigate the traditional value of Ewhaju (traditional wine) and to establish the brewing condition, studies on of traditional background and field inquiry were carried out. Scientific evaluation and possibility of revealation of Ewhaju were searched by the experiments of microbial and enzymatic properties of brewed Ewhaju and Nuruk by traditional method. In flora of microorganisms in Nuruk of Ewhaju, Aspergillus oryzae and Hansenula sP. were isolated, and, showed a level of 1.2$\times$$10^6$ CHU/g, respectiveln but other microorganisms were not grown in diluted cultivation test. The a-and f-amylase activity of Nuruk were 30.74 and 34.4, respectively and their activities of two amylases were 19.28 and 18.8 at first stage of brewing, 21.21 and 19.80 at 100 day after brewing, and 20.25 and 19.90 at one year aged Ewhaju, respectively. The brewed Ewhaju could be remained with high quality long period without teat treatment or addition of preservatives, also, stored Ewhaju contains remarka-bly high activity of amylases which might contribute to digestion.

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Properties of Lintnerized Waxy Rice Starches (산처리에 의한 찹쌀 전분의 성질 변화)

  • Park, Yang-Kyun;Kim, Sung-Kon;Kim, Kwan
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.22 no.5
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    • pp.596-602
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    • 1993
  • The Characteristics of Shinsunchalbyeo(Japonica) and Hangangchalbyeo($J{\times}Idica$) starches including physicochemical properties, differential scanning calorimetry(DSC) and enzymatic digestion of lintnerized starches were investigated. Degree of hydrolysis of Hangangchalbyeo starch with 2.2N HCI for 48hr was higher than that of shinsunchalbyeo starch. Absorbance at ${\lambda}_{max}$ 680nm, and ${\lambda}_{max}$ of iodine stained starch decreased upon acid treatment. But water binding capacity, swelling power and solubility considerably increased as hydrolysis progressed. Relative crystallinity of two starches increased with acid treatment, and that of Shinsunchalbyeo starch was higher than that of Hangangchalbyeo starch. DSC data continuously decreased for lintnerization periods, and those of Shinsunchalbyeo starch. DSC data continuously decreased for lintnerization periods, and those of Shinsunchalbyeo starch have higher than those of Hangangchalbyeo starch. The onset temperature of starch by DSC continuously decreased by treatment, but conclusion temperature increased until 24hr and then decreased. The enthalpy for gelatinization decreased for both starches. Degree of hydrolysis of lintnerized Shinsunchalbyeo starch with glucoamylase was slightly higher than that of Hangangchalbyeo starch.

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Inhibition of MMP-2 and MMP-9 activities by solvent-partitioned Sargassum horneri extracts

  • Karadeniz, Fatih;Lee, Seul-Gi;Oh, Jung Hwan;Kim, Jung-Ae;Kong, Chang-Suk
    • Fisheries and Aquatic Sciences
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    • v.21 no.6
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    • pp.16.1-16.7
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    • 2018
  • Background: Matrix metalloproteinases (MMPs) are linked with several complications such as metastasis of cancer progression, oxidative stress, and hepatic fibrosis. Brown seaweeds are being extensively studied for their bioactive molecule content against cancer progression. In this context, Sargassum horneri was reported to possess various bioactivities including antiviral, antimicrobial, and anti-inflammatory partly due to its phenolic compound content. Methods: In this study, potential of S. horneri was evaluated through anti-MMP effect in HT1080 fibrosarcoma cells. S. horneri crude extract was fractionated with organic solvents, namely, water ($H_2O$), n-buthanol (n-BuOH), 85% aqueous methanol (85% aq. MeOH), and n-hexane. The non-toxicity of fraction samples (Sargassum horneri solvent-partitioned extracts (SHEs)) was confirmed by cell-viability assay. SHEs were tested for their ability to inhibit MMP enzymatic activity through gelatin digestion evaluation and cell migration assay. Expressions of MMP-2 and MMP-9 and tissue inhibitors of MMP (TIMPs) were evaluated by reverse transcription and Western blotting. Results: All fractions inhibited the enzymatic activities of MMP-2 and MMP-9 according to gelatin zymography. Except $H_2O$ fraction, fractions hindered the cell migration significantly. All tested fractions suppressed both mRNA and protein levels of MMP-2, MMP-9, TIMP-1, and TIMP-2. Conclusion: Overall, current results suggested that S. horneri has potential to be a good source for anti-MMP agents, and further investigations are underway for better understanding of the action mechanism and isolation and elucidation of the bioactive molecules.

Composition and Diversity of Gut Bacteria Associated with the Eri Silk Moth, Samia ricini, (Lepidoptera: Saturniidae) as Revealed by Culture-Dependent and Metagenomics Analysis

  • MsangoSoko, Kondwani;Gandotra, Sakshi;Chandel, Rahul Kumar;Sharma, Kirti;Ramakrishinan, Balasubramanian;Subramanian, Sabtharishi
    • Journal of Microbiology and Biotechnology
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    • v.30 no.9
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    • pp.1367-1378
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    • 2020
  • The polyphagous eri silk moth, Samia ricini, is associated with various symbiotic gut bacteria believed to provide several benefits to the host. The larvae of S. ricini were subjected to isolation of gut bacteria using culture-dependent 16S rRNA generic characterization, metagenomics analysis and qualitative enzymatic assays. Sixty culturable aerobic gut bacterial isolates comprising Firmicutes (54%) and Proteobacteria (46%); and twelve culturable facultative anaerobic bacteria comprising Proteobacteria (92%) and Firmicutes (8%) were identified inhabiting the gut of S. ricini. The results of metagenomics analysis revealed the presence of a diverse community of both culturable and un-culturable gut bacteria belonging to Proteobacteria (60%) and Firmicutes (20%) associated with seven orders. An analysis of the results of culturable isolation indicates that these bacterial isolates inhabited all the three compartments of the gut. Investigation on persistence of bacteria coupled with metagenomics analysis of the fifth instar suggested that bacteria persist in the gut across the different instar stages. In addition, enzymatic assays indicated that 48 and 75% of culturable aerobic, and 75% of anaerobic gut bacterial isolates had cellulolytic, lipolytic and nitrate reductase activities, thus suggesting that they may be involved in food digestion and nutritional provision to the host. These bacterial isolates may be good sources for profiling novel genes and biomolecules for biotechnological application.

The review on standard method of microplastics in soil and groundwater (토양, 지하수 중 미세플라스틱 분석법에 관한 고찰)

  • JongBeom Kwon;Hyeonhee Choi;Sunhwa Park
    • Analytical Science and Technology
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    • v.37 no.3
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    • pp.174-188
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    • 2024
  • This review summarized research trends regarding sample collection methods, pretreatment method, and types of analysis devices for microplastics (MPs) in soil and groundwater matrices. Soil sampling considers the selection of sampling location, depth, and volume. The typically sampling depth is within 15 cm (topsoil), and about 1 kg of mixed each sample. Among spot sampling and continuous flow sampling, groundwater sampling mainly used a continuous flow sampling, with collection rates 2 to 6 L/min in the range of 300~1,000 L, and followed by immediate on-situ filtration. Pretreatment method, applied to soil and groundwater, consist of organic digestion and density separation. In the organic digestion method, H2O2 is recommended among H2O2, acidic, alkaline, and enzymatic method. NaCl is primarily used as a reagent in density separation. However, depending on the density of MPs, other regents can be selectively used like ZnCl2, ZnBr2, and etc. Representative analysis device includes Fourier Transform Infrared (FTIR) and Raman spectroscopy for non-destructive analysis and Pyrolysis Gas Chromatography Mass Spectrometry (Py-GC/MS) for destructive analysis. µ-FTIR and Raman can count MPs of larger than 10 and 1 ㎛, and analyze MPs materials. However, it is need to sufficiently remove interference, like organic matter, in spectroscopic analysis using essential pretreatment method. Py-GC/MS is being continuously researched because it doesn't require complex pretreatment method and allows quantitative analysis of specific materials.

Processing of Sardine Sauce from Sardine Scrap (정어리잔사를 이용한 정어리간장의 제조)

  • LEE Eung-Ho;CHO Soon-Yeong;HA Jae-Ho;OH Kwang-Soo;KIM Chang-Yang
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.17 no.2
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    • pp.117-124
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    • 1984
  • Sarine scrap usually comprises about $40\%$ of the raw fish in processing. The purpose of this study is to establish the desirable methods for proteinaceous materials from the sardine scrap through autolysis or enzymatic digestion and to convert them into useful by-products such as sardine sauce. Sardine scrap was chopped and mixed with equal weight of water, and be hydrolyzed them by autolysis and/or by addition of commercial proteolytic enzyme and various concentrations of sodium chloride. The optimal conditions for hydrolysis of sardine scrap were revealed in temperature at $55^{\circ}C$ and 4 hours digestion with bromelain($0.4\%$) and commercial complex enzyme ($6.0\%$), and those conditions were also applicated in autolysis. The maximum hydrolyzing rate of protein and released amino nitrogen were $82.5\%,\;5.2\%$ through autolysis, $84.3\%,\;5.8\%$ by bromelain digestion and $92.5\%,\;5.9\%$ by complex enzyme, respectively. In the products prepared from sardine scrap through autolysis or bromelain digestion, hypoxanthine was dominant, as $17.4 {\mu}mole/g$, dry matter for autolysis and $16.0 {\mu}mole/g$, dry matter, for bromelain digestion among the nucleotidcs and their related compounds, respectively. The abundant free amino acids were leucine, glutamic acid, lysnie, valine and alanine. The contents of those amino acids were $51.3\%,\;48.3\%$ of the total free amino acids, respectively. And the contents of 5'-IMP and TMAO were negligible but total creatinine was developed in value from $9.2\%\;to\;10\%$ of total extracted nitrogen. The flavor of sardine sauce prepared from sardine scrap by autolysis or enzyme digestion were not inferior to that of traditional Korean soy sauce by sensory evaluation.

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Molecular Weight and Structural Changes of Enzymatic Hydrolysate from Bombyx mori Fibroin (효소처리에 의한 Bombyx mori fibroin 가수분해물의 분자량 및 구조 변화)

  • Yeo, Joo-Hong;Park, Kyung-Ho;Lee, Kwang-Gill;Woo, Soon-Ok;Kweon, Hae-Yong;Han, Sang-Mi;Lee, Heui-Sam;Lee, Jin-Ah;Lee, In-Seok;Cho, Yun-Hi
    • Korean Journal of Food Science and Technology
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    • v.40 no.4
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    • pp.470-473
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    • 2008
  • This study examined the enzymatic digestion, average molecular weight distribution and structural changes of Bombyx mori silk gland fibroin using gel penneation chromatography and nuclear magnetic resonance methods. The pure-separation of calcium chloride-treated fibroin hydrolysates was carried out by gel filtration chromatography. Also, the effects of fibroin's enzymatic hydrolysis were investigated using an edible enzyme. The average molecular weight of three hydrolysate samples (silk gland fibroin (SF), SF-calcium chloride (SFC), and SFC-enzyme) were measured to compare their characteristics. The molecular weights of SF, SFC, and SFC-enzyme were approximately 108,000, 65,000, and 1,000 Da, respectively. Finally, we determined the structural characteristic changes of the different enzymatically digested samples by $^{13}C$ nuclear magnetic resonance methods. For the enzymatically digested fibroin, the glycine $^{13}C^{\alpha}$ resonance indicated that the amino acid was dramatically changed and/or separated out; however, this was not shown for the normal Bombyx mori silk gland fibroin.

BIOCHEMICAL CHARACTERIZATION OF EMBRYONIC CHICK CALVARIAL CELLS

  • Yu, Jae-Hyung;Kim, Jung-Kun;Cha, Kyung-Suk
    • The korean journal of orthodontics
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    • v.25 no.6 s.53
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    • pp.697-704
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    • 1995
  • Chicken calvarial bone is known to contain various cell types, but their exact composition is unknown. By characterizing the chicken calvarial bone biochemically, it can be used to study biochemical, histochemical actions of bone cells in general. Calvaria of 18-day-old white leg horn embryo was aseptically dissected and bone cell populations were isolated by sequential enzymatic digestion. Histochemical study for osteoclast-like bone cell. population was performed with tartrate resistant acid phosphatase(TRAP) stain and for osteoblast-like bone cell population, alkaline phosphatase(ALP) stain was performed. Biochemical study for osteoblast-like bone cell population was performed using alkaline phosphatase(ALP) assay. Following conclusions were obtained from this study. 1. TRAP positive multi and mononuclear cells were mostly observed in group I and II, indicating that osteoclast-like bone cell population is mostly found in these groups. 2. All the cultured groups showed almost equal ALP activities and were positive for ALP stain, indicating that osteoblast-like bone cell population is evenly dispersed in all culture groups. 3. Experimental group treated with $1,25(OH)_{2}D_3$ showed increase in ALP activity in contrast to the control group, confirming previous studies that $1,25(OH)_{2}D_3$ increases ALP activities in in vitro bone cultures. 4. Results from von Kossa's stain indicated that in vitro bone formation had occured after 3 weeks of culture with beta-glycero phosphate.

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Effects of a ${\delta}-opioid$ Agonist on the Brainstem Vestibular Nuclear Neuronal Activity of Rats

  • Kim, Tae-Sun;Huang, Mei;Jang, Myung-Joo;Jeong, Han-Seong;Park, Jong-Seong
    • The Korean Journal of Physiology and Pharmacology
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    • v.9 no.3
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    • pp.137-141
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    • 2005
  • This study was undertaken to investigate the effects of [$D-Ala^2$, D-Leu^5$]-enkephalin (DADLE) on the spontaneous activity of medial vestibular nuclear neurons of the rat. Sprague-Dawley rats, aged 14 to 16 days, were anesthetized with ether and decapitated. After enzymatic digestion, the brain stem portion of medial vestibular nuclear neuron was obtained by micropunching. The dissociated neurons were transferred to a recording chamber mounted on an inverted microscope, and spontaneous action potentials were recorded by standard patch-clamp techniques. The spontaneous action potentials were increased by DADLE in 12 cells and decreased in 3 cells. The spike frequency and resting membrane potential of these cells were increased by DADLE. The depth of afterhyperpolarization was not affected by DADLE. The potassium currents were decreased in 20 cells and increased in 5 cells. These results suggest that DADLE increases the neuronal activity of the medial vestibular nuclear neurons by altering resting membrane potential.