• Title/Summary/Keyword: Kinetic Parameters

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Purification and Characterization of Protease Produced by Aspergillus wentti Isolated from Korean Traditional Meju (한국 전통 메주 유래의 Aspergillus wentti가 생성하는 Protease 의 정제 및 특성)

  • Lim, Seong-Il
    • Korean Journal of Food Science and Technology
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    • v.32 no.1
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    • pp.161-167
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    • 2000
  • The protease produced by a newly isolated Aspergillus wentti from Korean traditional Meju was purified and characterized. The optimal medium composition and culture conditions for maximum protease production were ; bran :1% glucose solution =1 : 1, pH 9.0, $30^{\circ}C$, and 4 days of fermentation. Protease was purified by QAE-Sephadex, SP-Sephadex ion exchange chromatography and Sephadex G-100 chromatography. The specific activity and the purification fold of the purified enzyme were 213 unit/mg protein and 27.3, respectively. The molecular weight of purified protease was found to be 32 kDa by SDS-PAGE. Km and Vmax value's for hammastein milk casein were $3.049{\times}10^{-4}\;M\;and\;151.1\;{\mu}g/min$, respectively. Kinetic parameters showed that the enzyme has higher affinity to casein than isolated soybean protein, hemoglobin and bovine serum albumin. Optimal pH and temperature for reaction of the purified enzyme were 9.0 and $50^{\circ}C$, respectively. The enzyme was stable at pH 4.0-11.0, below $40^{\circ}C$, and the activity was not stimulated by metal ions. 1mM phenylmethylsulfonyl fluoride inhibited the enzyme activity by 98.5%. It means that the enzyme is one of serine protease.

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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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Kinetics of Thermal Inactivation of Peroxidases and Polyphenol Oxidase in Pineapple (Ananas comosus)

  • Lee, Ting Hun;Chua, Lee Suan;Tan, Eddie Ti Tjih;Yeong, Christina;Lim, Chew Ching;Ooi, Siew Yin;Aziz, Ramlan bin Abdul;Aziz, Azila binti;Sarmidi, Mohd Roji bin
    • Food Science and Biotechnology
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    • v.18 no.3
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    • pp.661-666
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    • 2009
  • The heat tolerance and the inactivation kinetics of peroxidase (POD) and polyphenol oxidase (PPO) in pineapples (Ananas comosus) were studied in the temperature range $45-95^{\circ}C$. The kinetic parameters, such as deactivation rate constant (k), activation energy ($E_a$), and decimal reduction rate (D) of the thermal inactivation process, were determined. POD in pineapples showed biphasic inactivation behavior at temperatures range $45-75^{\circ}C$ but was monophasic at $85-95^{\circ}C$. This indicate that POD has 2 isozymes, namely heat labile and heat resistant, with $E_a$ of 68.79 and 93.23 kJ/mol, respectively. On the other hand, the heat denaturation of pineapple PPO could be described as simple monophasic first-order behavior with $E_a$ of 80.15 kJ/mol. Thus, the results of this study is useful in blanching technology where it shows a shortened time with higher temperature can be applied. The determination of the heat tolerance and inactivation POD and PPO, at different temperature range as done in the present work, was very important to improve the blanching process. This also will help to optimize the pineapple canning process which is one of the most important food industries in many tropical regions.

L-Lysine Production by 6-Azauracil Resistant Mutant of Corynebacterium glutamicum (6-Azaumcil 내성을 지닌 Corynebacterium glutamicum 변이주에 의한 L-Lysine의 생산)

  • 신현철;김성준전영중이재흥
    • KSBB Journal
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    • v.9 no.4
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    • pp.372-377
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    • 1994
  • To improve L-lysine yield, pyrimidine base analogue(6-azauracil)-resistant mutants were isolated from Corynebacterium glutamicum KFCC10672 Among them the best producer, C. glutamicum CH0516, was selected and tested for L-lysine production in a $7\ell$ fermentor. It was found that the product yield obtained with C. glutamicum CH0516 was higher than that of the parent strain by 3%. In order to elucidate the gain in productivity with the 6-azauracil-resistant mutant enzymatic kinetic parameters such as aspartokinase(AKase) and aspartate carbamoyltransferase (ATCase) were measured. The Km values of AKase with C. glutamicum KFCC10672 and CH0516 were 200.0 mM and 166.7 mM and those of ATCase were 0.13 mM and 0.27 mM, respectively. However, the specific enzyme activities of AKase of C. glutamlcum KFCC10672 and CH0516 were $3.89{\times}10^{-1}$ units/mg and $4.78{\times}10^{-1}$ units/mg, and those of ATCarse were 2.20 units/mg and 1.84 units/mg, respectively. It appears that some increase in product yield with C. gluramicum CH0516 is likely due to the increased Akase activity and decreased ATCase activity.

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Gene Cloning, Purification and Characterization of Xylanase 10A from Paenibacillus woosongensis in Escherichia coli (Paenibacillus woosongensis로부터 대장균에 Xylanase 10A의 유전자 클로닝과 정제 및 특성분석)

  • Yoon, Ki-Hong
    • Microbiology and Biotechnology Letters
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    • v.48 no.2
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    • pp.158-166
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    • 2020
  • A gene coding for the xylanase was cloned from Paenibacillus woosongensis, followed by determination of its complete nucleotide sequence. This xylanase gene, designated as xyn10A, consists of 1,446 nucleotides encoding a polypeptide of 481 amino acid residues. Based on the deduced amino acid sequence, Xyn10A was identified to be a modular enzyme composed of a catalytic domain highly homologous to the glycosyl hydrolase family 10 xylanase and a putative carbohydrate-binding module (CBM) in the C-terminus. By using DEAE-sepharose and phenyl-sepharose column chromatography, Xyn10A was purified from the cellfree extract of recombinant Escherichia coli carrying a P. woosongensis xyn10A gene. The N-terminal amino acid sequence of the purified Xyn10A was identified to exactly match the sequence immediately following the signal peptide predicted by the Signal5.0 server. The purified Xyn10A was a truncated protein of 33 kDa, suggesting the deletion of CBM in the C-terminus by intracellular hydrolysis. The purified enzyme had an optimum pH and temperature of 6.0 and 55-60℃, respectively, with the kinetic parameters Vmax and Km of 298.8 U/mg and 2.47 mg/ml, respectively, for oat spelt xylan. The enzyme was more active on arabinoxylan than on oat spelt xylan and birchood xylan with low activity for p-nitrophenyl-β-xylopyranoside. Xylanase activity was significantly inhibited by 5 mM Cu2+, Mn2+, and SDS, and was noticeably enhanced by K+, Ni2+, and Ca2+. The enzyme could hydrolyze xylooligosaccharides larger than xylobiose. The predominant products resulting from xylooligosaccharide hydrolysis were xylobiose and xylose.

Prediction of Axillary Lymph Node Metastasis in Early Breast Cancer Using Dynamic Contrast-Enhanced Magnetic Resonance Imaging and Diffusion-Weighted Imaging

  • Jeong, Eun Ha;Choi, Eun Jung;Choi, Hyemi;Park, Eun Hae;Song, Ji Soo
    • Investigative Magnetic Resonance Imaging
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    • v.23 no.2
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    • pp.125-135
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    • 2019
  • Purpose: The purpose of this study was to evaluate dynamic contrast-enhanced breast magnetic resonance imaging (DCE-MRI), and diffusion-weighted imaging (DWI) variables, for axillary lymph node (ALN) metastasis in the early stage of breast cancer. Materials and Methods: January 2011-April 2015, 787 patients with early stage of breast cancer were retrospectively reviewed. Only cases of invasive ductal carcinoma, were included in the patient population. Among them, 240 patients who underwent 3.0-T DCE-MRI, including DWI with b value 0 and $800s/mm^2$ were enrolled. MRI variables (adjacent vessel sign, whole-breast vascularity, initial enhancement pattern, quantitative kinetic parameters, signal enhancement ratio (SER), tumor apparent diffusion coefficient (ADC), peritumoral ADC, and peritumor-tumor ADC ratio) clinico-pathologic variables (age, T stage, multifocality, extensive intraductal carcinoma component (EIC), estrogen receptor, progesterone receptor, HER-2 status, Ki-67, molecular subtype, histologic grade, and nuclear grade) were compared between patients with axillary lymph node metastasis and those with no lymph node metastasis. Multivariate regression analysis was performed, to determine independent variables associated with ALN metastasis, and the area under the receiver operating characteristic curve (AUC), for predicting ALN metastasis was analyzed, for those variables. Results: On breast MRI, moderate or prominent ipsilateral whole-breast vascularity (moderate, odds ratio [OR] 3.45, 95% confidence interval [CI] 1.28-9.51 vs. prominent, OR = 15.59, 95% CI 2.52-96.46), SER (OR = 1.68, 95% CI 1.09-2.59), and peritumor-tumor ADC ratio (OR = 6.77, 95% CI 2.41-18.99), were independently associated with ALN metastasis. Among clinico-pathologic variables, HER-2 positivity was independently associated, with ALN metastasis (OR = 23.71, 95% CI 10.50-53.54). The AUC for combining selected MRI variables and clinico-pathologic variables, was higher than that of clinico-pathologic variables (P < 0.05). Conclusion: SER, moderate or prominent increased whole breast vascularity, and peritumor-tumor ADC ratio on breast MRI, are valuable in predicting ALN metastasis, in patients with early stage of breast cancer.

Bio Toxicity Assessment and Kinetic Model of 6 Heavy Metals Using Luminous Bacteria (발광미생물을 이용한 중금속 6종의 생물독성 평가 및 모델링)

  • Kim, Ilho;Lee, Jaiyeop
    • Journal of the Korean Society of Urban Environment
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    • v.18 no.4
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    • pp.547-555
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    • 2018
  • In addition to North America and Europe, Korea is also responding to the toxic damage caused by the production and distribution of chemicals. Methods for assessing bio-toxicity of harmful substances have been widely introduced, but it is required of quantitative and speedy information for modeling. For 6 heavy metals, as zinc, copper, chrome, cadmium, mercury and lead, bio-toxicity assessment and kinetics model were constructed using Vibrio fischeri which is widely used luminous bacteria. The degree of luminescence activity and the toxicity of heavy metals were relative limunescence unit, RLU measured as by using a photomultiplier embedded device. The toxicity was assessed by the concentration levels giving under 20% lethality and lethal concentration, $EC_{50}$. In the results, the toxicity order were followed from mercury, lead, copper, chrome, zinc and cadmium. $EC_{{50},{\infty}}$ obtained by trends of $EC_{50}$ by time follows had highly linear agreement with main parameters of bio-toxicity modelling. The average error rates of the reproduced lethality obtained from DAM and TDM model on the basis of body residue, were 10.2% for mercury, lead, copper, chrome and 20.0 for the all 6 methals.

Risk assessment for norovirus foodborne illness by raw oyster (Ostreidae) consumption and economic burden in Korea

  • Yoo, Yoonjeong;Oh, Hyemin;Lee, Yewon;Sung, Miseon;Hwang, Jeongeun;Zhao, Ziwei;Park, Sunho;Choi, Changsun;Yoon, Yohan
    • Fisheries and Aquatic Sciences
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    • v.25 no.5
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    • pp.287-297
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    • 2022
  • The objective of this study was to evaluate the probability of norovirus foodborne illness by raw oyster consumption. One hundred fifty-six oyster samples were collected to examine the norovirus prevalence. The oyster samples were inoculated with murine norovirus and stored at 4℃-25℃. A plaque assay determined norovirus titers. The norovirus titers were fitted with the Baranyi model to calculate shoulder period (h) and death rate (Log PFU/g/h). These kinetic parameters were fitted to a polynomial model as a function of temperature. Distribution temperature and time were surveyed, and consumption data were surveyed. A dose-response model was also searched through literature. The simulation model was prepared with these data in @RISK to estimate the probability of norovirus foodborne. One sample of 156 samples was norovirus positive. Thus, the initial contamination level was estimated by the Beta distribution (2, 156), and the level was -5.3 Log PFU/g. The developed predictive models showed that the norovirus titers decreased in oysters under the storage conditions simulated with the Uniform distribution (0.325, 1.643) for time and the Pert distribution (10, 18, 25) for temperature. Consumption ratio of raw oyster was 0.98%, and average consumption amount was 1.82 g, calculated by the Pert distribution [Pert {1.8200, 1.8200, 335.30, Truncate (0, 236.8)}]. 1F1 hypergeometric dose-response model [1 - (1 + 2.55 × 10-3 × dose)-0.086] was appropriate to evaluate dose-response. The simulation showed that the probability of norovirus foodborne illness by raw oyster consumption was 5.90 × 10-10 per person per day. The annual socioeconomic cost of consuming raw oysters contaminated with norovirus was not very high.

Kinetic and Statistical Analysis of Adsorption and Photocatalysis on Sulfamethoxazole Degradation by UV/$TiO_2$/HAP System (UV/$TiO_2$/HAP 시스템에서 Sulfamethoxazole의 흡착과 광촉매반응에 대한 동역학적 및 통계적 해석)

  • Chun, Suk-Young;Chang, Soon-Woong
    • Journal of the Korean GEO-environmental Society
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    • v.13 no.5
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    • pp.5-12
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    • 2012
  • Antibiotics have been considered emerging compounds due to their continuous input and persistence in environment. Due to the limited biodegradability and widespread use of these antibiotics, an incomplete removal is attained in conventional wastewater treatment plants and relative large quantities are released into the environment. In this study, it was determined the adsorption and photocatalysis kinetics of antibiotics (Sulfamethoxazole, SMX) with various catalyst (Titanium dioxide; $TiO_2$, Hydroxyapatite; HAP) conditions under UV/$TiO_2$/HAP system. In addition, the statistical analysis of response surface methods (RSM) was used to determine the effects of operating parameters on UV/$TiO_2$/HAP system. $TiO_2$/HAP adsorbent were found to follow the pseudo second order reaction in the adsorption. In the result of applied intrapaticle diffusion model, the constants of reaction rate were $TiO_2$=$0.064min^{-1}$, HAP=$0.2866min^{-1}$ and $TiO_2$/HAP=$0.3708min^{-1}$, respectively.The result of RSM, term of regression analysis in analysis of variance (ANOVA) showed significantly p-value (p<0.05) and high coefficients for determination values($R^2$=96.2%, $R^2_{Adj}$=89.3%) that allowed satisfactory prediction of second order regression model. And the estimated optimal conditions for Y(Sulfamethoxazole removal efficiency, %) were $x_1$(initial concentration of Sulfamethoxazole)=-0.7828, $x_2$(amount of catalyst)=0.9974 and $x_3$(reation time)=0.5738 by coded parameters, respectively. According to the result of intraparticle diffusion model and photocatalysis experiments, it was shown that the $TiO_2$/HAP was more effective system than conventional AOPs(advanced oxidation processes, UV/$TiO_2$ system).

Comparison of Algorithms for Generating Parametric Image of Cerebral Blood Flow Using ${H_2}^{15}O$ PET Positron Emission Tomography (${H_2}^{15}O$ PET을 이용한 뇌혈류 파라메트릭 영상 구성을 위한 알고리즘 비교)

  • Lee, Jae-Sung;Lee, Dong-Soo;Park, Kwang-Suk;Chung, June-Key;Lee, Myung-Chul
    • The Korean Journal of Nuclear Medicine
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    • v.37 no.5
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    • pp.288-300
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    • 2003
  • Purpose: To obtain regional blood flow and tissue-blood partition coefficient with time-activity curves from ${H_2}^{15}O$ PET, fitting of some parameters in the Kety model is conventionally accomplished by nonlinear least squares (NLS) analysis. However, NLS requires considerable compuation time then is impractical for pixel-by-pixel analysis to generate parametric images of these parameters. In this study, we investigated several fast parameter estimation methods for the parametric image generation and compared their statistical reliability and computational efficiency. Materials and Methods: These methods included linear least squres (LLS), linear weighted least squares (LWLS), linear generalized least squares (GLS), linear generalized weighted least squares (GWLS), weighted Integration (WI), and model-based clustering method (CAKS). ${H_2}^{15}O$ dynamic brain PET with Poisson noise component was simulated using numerical Zubal brain phantom. Error and bias in the estimation of rCBF and partition coefficient, and computation time in various noise environments was estimated and compared. In audition, parametric images from ${H_2}^{15}O$ dynamic brain PET data peformed on 16 healthy volunteers under various physiological conditions was compared to examine the utility of these methods for real human data. Results: These fast algorithms produced parametric images with similar image qualify and statistical reliability. When CAKS and LLS methods were used combinedly, computation time was significantly reduced and less than 30 seconds for $128{\times}128{\times}46$ images on Pentium III processor. Conclusion: Parametric images of rCBF and partition coefficient with good statistical properties can be generated with short computation time which is acceptable in clinical situation.