• Title/Summary/Keyword: Enzymatic pattern

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Isotope-Dilution Mass Spectrometry for Quantification of Urinary Active Androgens Separated by Gas Chromatography

  • Lee, Su-Hyeon;Choi, Man-Ho;Lee, Won-Yong;Chung, Bong-Chul
    • Mass Spectrometry Letters
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    • v.1 no.1
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    • pp.29-32
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    • 2010
  • Cross reacting antibodies can cause an overestimation of the results of immunoassays. Therefore, alternative methods are needed for the accurate quantification of steroids. Gas chromatography combined with isotope-dilution mass spectrometry (GC-IDMS) is developed to quantify urinary active androgens, testosterone, epitestosterone and dihydrotestosterone, which are clinically relevant androgens to both hair-loss and prostate diseases. The method devised involves enzymatic hydrolysis with $\beta$-glucuronidase, solid-phase extraction, liquid-liquid extraction using methyl tert-butyl ether and subsequent conversion to pentafluorophenyldimethylsilyl-trimethylsilyl (flophemesyl-TMS) derivatives for sensitive and selective analysis in selected-ion monitoring mode. Flophemesyl-TMS derivatization not only eliminates matrix interference but also has a good peak resolution within a 6 min-run. A selective and sensitive GC technique with flophemesyl-TMS derivatives also allows accurate quantitative analysis of three active androgens when combined with IDMS. The limit of quantification of the three analytes was <50 pg/mL, and extraction recoveries ranged from 91.9 to 102.1%. The precision and accuracy were 1.2~6.5% and 89.0~106.7%, respectively. This GC-IDMS method can be useful for evaluating the drug efficacy and monitoring the biological processes responsible for male-pattern baldness and prostate diseases.

한국동물학회 제 14회 대회기록: 유전학의 제문제

  • Ojima, Yoshio
    • The Korean Journal of Zoology
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    • v.13 no.4
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    • pp.112-126
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    • 1970
  • This paper deals with cytogenetical and cytochemical studies of the carp (Cyprinus carpio), the funa (Carassius carassius) and their hybrids. When kept under a confined condition, the carp and the funa mate andcan produce hybrids. Reciprocal crosses are also possible with similar results. The hybrids grow regularly with no observed abnormalities in the course of their development. They rank intermediate between the parent species in several characters. The hybrid males are completely sterile, while a hybrid female laid eggs in backcrossing. The spermatogenetic activity in hybrid testes is greatly disturbed. The chromosomes as observed in spermatogonial devision of hybrids are 100 in number, being the total sum of the haploid numbers of the parents, 50 for the carp and 50 for the funa. Meiosis in the hybrid testes is highly disturbed being arrested at early stages of the meiotic prophase. Most of the germ-cells undergo pycnotic degeneration during the period from late leptotene, and no spermatozoa are produced. In some hybrid specimens, the gonads show mosaic structures composed of testicular and ovarian elements, anevidence suggesting that sterility is associated with intersexuality caused by genetic unbalance between the parent species. The DNA amount in spermatogonial nuclei of thehybrids is approximately the same as that of liver nuclei, showing the 2n value. The DNA amount in the pachytene nuclei of the hybrids is less than the 4n value, while the parent species have the reduced amount of DNA in their pachytene nuclei. A consideration was made that the reduced amount of DNA in the hybrid cells may cause the disturbance of cellular activity leading to the subsequent degeneration of cells. Some aspects of enzymatic pattern in the carp, funa and their hybrids are. going on.

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Physicochemical Property Changes of Sweet Potato Starch by Ultra Fine Pulverization

  • Kim, Hee-Sun;Park, Hye-Young;Han, Gwi-Jung;Kim, Myung-Hwan
    • Food Engineering Progress
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    • v.15 no.2
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    • pp.169-174
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    • 2011
  • This study was performed to analyze the effects of ultra fine pulverization (UFP) on the physicochemical properties of sweet potato starch (SPS). The average diameter and specific surface area of the SPS was decreased from 22.94 to 10.25 $\mu$m and from 0.879 to 1.909 $m^2$ /g throughout UFP, respectively, and the damaged starch content was increased from 13.7 to 99.2%. The pulverized sweet potato starch (PSPS) had higher swelling power, solubility, and transmittance values than the SPS. X-ray diffractograms revealed that the SPS had a C-type pattern, which disappeared in PSPS. The rapid visco analysis (RVA) characteristics, peak viscosity, break down, and set back of SPS ceased to exist in PSPS. According to differential scanning calorimetry (DSC) curves, the peak temperature ($T_p$) and gelatinization enthalpy ($\Delta$E) of SPS were $71.95^{\circ}C$ and 10.40 J/g, respectively, while these remained undetected in PSPS. The enzymatic digestibilities of SPS and PSPS were 61.7 and 84.7%, respectively.

Varying Inocula Permutations (Aspergillus oryzae and Bacillus amyloliquefaciens) affect Enzyme Activities and Metabolite Levels in Koji

  • Gil, Hye Jeong;Lee, Sunmin;Singh, Digar;Lee, Choong Hwan
    • Journal of Microbiology and Biotechnology
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    • v.28 no.12
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    • pp.1971-1981
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    • 2018
  • In this study, we investigated the altered enzymatic activities and metabolite profiles of koji fermented using varying permutations of Aspergillus oryzae and/or Bacillus amyloliquefaciens. Notably, the protease and ${\beta}$-glucosidase activities were manifold increased in co-inoculated (CO) koji samples (co-inoculation of A. oryzae and B. amyloliquefaciens). Furthermore, gas chromatography-mass spectrometry (GC-MS)-based metabolite profiling indicates that levels of amino acids, organic acids, sugars, sugar alcohols, fatty acids, nucleosides, and vitamins were distinctly higher in CO, SA (sequential inoculation of A. oryzae, followed by B. amyloliquefaciens), and SB (sequential inoculation of B. amyloliquefaciens, followed by A. oryzae). The multivariate principal component analysis (PCA) plot based on GC-MS datasets indicated a clustered pattern for MA and MB (koji samples inoculated either with A. oryzae or B. amyloliquefaciens) across PC2 (20.0%). In contrast, the CO, SA, and SB metabolite profiles displayed segregated patterns across PLS1 (22.2%) and PLS2 (21.1%) in the partial least-square discriminant analysis (PLS-DA) model. Intriguingly, the observed disparity in the levels of primary metabolites was engendered largely by higher relative levels of sugars and sugar alcohols in MA, SA, and CO koji samples, which was commensurate with the relative amylase activities in respective samples. Collectively, the present study emphasizes the utility of integrated biochemical and metabolomic approaches for achieving the optimal permutation of fermentative inocula for industrial koji preparation.

Biological Potential of Novel Specific Casein-Derived Peptides

  • Kim, Da Young;Yoo, Jung Sik;Cho, Yoon Ah;Yoon, Ho Sik;Kim, Cheol-Hyun
    • Journal of Dairy Science and Biotechnology
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    • v.39 no.1
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    • pp.36-50
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    • 2021
  • This study aimed to evaluate the biological potential of functional food, i.e., specific peptides obtained from the hydrolysis of milk protein, by assessing their antioxidant and antibacterial properties. For the preparation of casein hydrolysates, commercial enzymes were added to 10% casein solution in a 1:200 (w/v) ratio, and samples were collected each hour. Based on the assessment of the degree of hydrolysis (DH) of casein hydrolysates, it was observed that the concentration of all enzymatic hydrolysates increased rapidly from 30 to 40 minutes. However, no change was observed in their concentrations after 150 minutes. Protamex® and Neutrase® exhibited the highest DH when compared to other enzymes. Furthermore, SDS-PAGE was performed for analyzing the proteolytic pattern of each enzyme, except for Flavourzyme®, and peptides in the size range of 20-25 kDa were identified. Subsequently, peptides produced by two enzymes were isolated using a preparative liquid chromatography system. Overall, NF3, NF4, PF5, and PF6 showed higher antioxidant potential than other peptide fractions. Moreover, NF7 and PF3 exhibited the highest antibacterial activity. In this study, we evaluated the biological potential of novel casein-derived peptides that may find application in the food and healthcare industry.

The Correlativity of Ulva lactuca Fractions, LPS, Enzymatic Activity and the Evaluation of Water Fraction (갈파래 분획, LPS, 효소활성의 상관성과 수층분획의 가치평가)

  • Nam, Chun-Suk;Kang, Kum-Suk;Ha, Jong-Myung;Lee, Sang-Hyeon;Lee, Jae-Hwa;Lee, Dong-Geun;Jang, Jeong-Su;Kang, Hwan-Yul;Ha, Bae-Jin
    • Journal of Life Science
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    • v.16 no.6
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    • pp.984-988
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    • 2006
  • Lipopolysaccharide(LPS) was posttreated after the 14 day-pretreatment of Ulva lactuca fractions(ULF), and their correlativity to enzymatic activity alteration was investigated in the liver of rats. ULF was intraperitoneally administered into rats at dose of $1m{\ell}/kg$ of 100 mg/kg concentration for 14 days. On the day 15, $1m{\ell}/kg$ of LPS was injected. The corelativity was examined by measuring the changed values of superoxide dismutase(SOD) in mitochondrial fraction and catalase(CAT), glutathione peroxidase(GPx) in liver homogenate. The results showed that LPS treatment decreased the high values of SOD, CAT, GPx to the low values, but ULF pretreatment increased the low values of SOD, CAT, GPx to the high values. It was suggested that ULF, LPS and antioxidative enzymes like SOD, CAT, GPx had the corelativity of the high-low-high pattern and that the ULF pretreatment played the proper preventive role in the protection against the LPS treatment-induced enzymatic inactivity in the water fraction.

Cognitive Ability Enhancement Effects in Rats by B. mori Fibroin Enzymatic Hydrolysate (견 피브로인 효소 가수분해물의 동물 인지기능 향상 효과)

  • Yeo Joo-Hong;Lee Kwang-Gill;Kweon HaeYong;Woo Soon-Ok;Han Sang-Mi;Lee Yong-Woo;Kim Jin-Il;Kim Sung-Su;Demura Makoto
    • Journal of Sericultural and Entomological Science
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    • v.46 no.1
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    • pp.23-27
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    • 2004
  • We were investigated the cognitive ability enhancement in rats using B. mori fibroin molecular controlled hydrolysate by preparative recycle HPLC system. Also, some of its physicochemical properties and free amino acid components were investigated, too. Nuclear magnetic resonance (NMR) spectrum are measured different pattern between high (HF) and low (LF) molecular controlled B. mori fibroin samples, respectively. The in vitro test by neuron primary cell culture are showed dependent on molecular weight (Mw) of B. mori fibroin in the order of LF > MF > HF, too. However, the memorial effects in rats by in vivo test are large dependent molecular weight on that maximum 50% than control. Also, the memorial effect was higher than the enzymatic relative and acid hydrolysate, which was 50.0$\pm$2.1 and 25.9$\pm$0.32%, respectively.

Derivatization of Rice Wine Meal Using Commercial Proteases and Characterization of Its Hydrolysates (단백질가수분해효소를 이용한 탁주박의 가수분해 및 그 분해물의 특성)

  • Kim, Chang-Won;Choi, Hyuk-Joon;Han, Bok-Kyung;Yoo, Seung-Seok;Kim, Chang-Nam;Kim, Byung-Yong;Baik, Moo-Yeol
    • Korean Journal of Food Science and Technology
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    • v.43 no.6
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    • pp.729-734
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    • 2011
  • With the goal of transforming rice protein from an insoluble to a soluble form to increase the industrial utilization of rice wine meal (RWM), RWM was derivatized using commercial proteases and the RWM hydrolysates were characterized. Eight commercial proteases were used individually or in combination for hydrolysis of RWM. The degree of hydrolysis was assessed by determining the soluble protein in supernatant using the Lowry assay, protein in precipitates using a semimicro Kjeldahl procedure, and gravimetrically by the weight difference before and after hydrolysis. Protamex, Alcalase and Protease N proteases were most effective for hydrolysis of RWM. Although these assessment methodologies displayed some variation, they generally showed a similar pattern. When the aforementioned three proteases were simultaneously used to treat RWM, no significant difference was observed between the three assays (p<0.05) indicating an absence of enzymatic synergy.

Functional Activities of Low Molecular Weight Peptides Purified from Enzymatic Hydrolysates of Seaweeds (해조류 효소가수분해물질로부터 정제한 저분자 Peptide의 기능성)

  • Lee, Jung-Min;You, Sang-Guan;Kim, Sang-Moo
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.34 no.8
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    • pp.1124-1129
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    • 2005
  • Functional activities of low molecular weight substances purified from pepsin hydrolysates of four different seaweeds; Costaria costata, Enteromorpha prolifera, Grateloupia filicina and Porphyra tenera, were inves-tigated. Each pepsin hydrolysate of Costaria costata, Enteromorpha prolifera, and Grateloupia filicina resulted in three peptide peaks on Bio-Rad P2 gel chromatography pattern, while that of Porphyra tenera showed 2 peaks. Peak 1 of Porphyra tenera showed the highest antioxidative activity followed by peak 2 of Porphyra tenera and peak 2 of Costaria costata in order Peak 1 of Porphyra tenera showed the highest ACE inhibitory activity followed by peak 3 and peak 2 of Enteromorpha prolifera in order. Peak 1 and peak 2 of Porphyra tenera, and peak 2 of Enteromorpha prolifera showed the highest antityrosinase activity followed by peak 3 of Enteromorpha prolifera. Peak 1 of Enteromorpha prolifera showed the highest antitumor activity followed by peak 2 of Costaria costata, peak 3 of Enteromorpha prolifera, and peak 3 of Grateloupia filicina in order. Porphyra tenera showed the highest functional activities, which is thought to be due to its high protein content. Structure and amino acid sequence of low molecular weight peptide of Porphyra tenera should be analyzed in the further study.

Enzymatic Modification of Soy Proteins: Effects of Functional Properties of Soy Isolate upon Proteolytic Hydrolysis (대두단백질(大豆蛋白質)의 효소적(酵素的) 변형(變形) : 분리대두단백질(分離大豆蛋白質)의 기능성(機能性)에 미치는 단백질가수분해(蛋白質加水分解)의 영향(影響))

  • Kang, Yeung-Joo
    • Korean Journal of Food Science and Technology
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    • v.16 no.2
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    • pp.211-217
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    • 1984
  • To study affinity of proteolytic enzymes to soy proteins, the physicochemical and functional properties of enzymatically modified protein products, kinetic parameters and degree of hydrolysis were measured using trypsin, alcalase (serine type protease) and pronase. Bacterial alcalase and pronase showed much greater affinity to soy protein than animal intestinal trypsin. This effect was very significant when unheated soy isolate was used as a substrate. Specific activities of these enzymes decreased with the increment of substrate concentration (over 2.0%, w/v) when heat denatured soy protein was used as a substrate. However, the decrease in specific activity was negligible at substrate concentrations lower than 2.0%. Polyacrylamide gel electrophoretic results showed that the pattern of 2S protein band changed distinctly in alcalase hydrolysis as compared with those of trypsin and pronase. Protein solubilities of alcalase and pronase hydrolyzates increased by 25-30%, at their pI (pH 5.0) over the control. Virtually no change was observed in solubility by trypsin hydrolysis. Heat coagulability and calcium-tolerance of the protein increased by enzymatic hydrolysis. No clear tendency, however, was observed for emulsion properties, foam expansion and the amount of free -SH groups. The enzyme treatment considerably decreased foam stability.

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