• Title/Summary/Keyword: Enzyme Profile

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Characterization of Polyphenoloxidase extracted from Solanum tuberosum Jasim

  • Jang, Jae-Won;Ma, Yu-Hyun;Shin, Ju-Mi;Song, Kyung-Bin
    • Food Science and Biotechnology
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    • v.14 no.1
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    • pp.117-122
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    • 2005
  • Polyphenoloxidase (PPO) was extracted from Solanum tuberosum Jasim by various chromatographic methods and was subsequently purified and characterized. PPO was purified upto 78-fold from the crude extract. SDS-PAGE profile of the enzyme showed a major subunit of PPO with molecular weight of 40 kDa. The optimum pH and temperature for the maximum activity of PPO was 6.5 and $25^{\circ}C$, respectively. The enzyme was found to be quite stable between 10 and $40^{\circ}C$, whereas it was almost inactivated at $70^{\circ}C$ when incubated for 30 min. Substrate specificity study indicated that catechol was the most suitable substrate for PPO isolated from purple-fleshed potato with a $K_m$ value of 21.1 mM. The most effective inhibitor was ascorbic acid, followed by L-cysteine, citric acid, EDTA, and boric acid. Studies on the effect of metal ion on PPO activity showed that magnesium and copper were inhibitory, while iron and zinc ions increased the activity of PPO.

Genetic Diversity of Korean Cyanobacteria determined by DNA polymorphisms within the Phycocyanin Locus (Phycocyanin locus내의 DNA Polymorphism에 의한 한국산 Cyanobacteria의 유전적 다양성)

  • 박진숙;권주리;유순애
    • Korean Journal of Microbiology
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    • v.36 no.4
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    • pp.249-253
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    • 2000
  • The genetic diversity among Korean cyanobacteria was assessed by restriction fragment length polymorphism(RFLP) analysis of PCR products from the phycocyanin locus. Strains of all the genera tested were successfully amplified, and the size of amplified fragments was approximately 700bp. The restriction patterns generated by AluI, MspI, and HaeIII were conserved for strains within each of genera studied and were specific to the genus level. Intrageneric delineation of strains was revealed by the enzyme, CfoI for members of genera Anabeana and Synechocystis. Phenogram derived from the different RFLP patterns revealed a coherent cluster among Anabeana, Chlorogloea, and Synechocystis strains. PC-RFLP methods provided useful tools for classification of cyanobacteria.

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Long-term Tolvaptan Treatment of Autosomal Dominant Polycystic Kidney Disease in Korea

  • Kim, Ha Yeon;Lee, Seung Jin;Kim, Byung Ki;Kim, Minah;Bae, Eun Hui;Ma, Seong Kwon;Kim, Soo Wan
    • Electrolytes & blood pressure
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    • v.16 no.2
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    • pp.23-26
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    • 2018
  • A 22-year-old male patient was diagnosed with autosomal dominant polycystic kidney disease (ADPKD). He received conservative treatment with an angiotensin-converting enzyme inhibitor. Two years later, oral therapy, consisting of 60 mg tolvaptan per day, was initiated. Compared with height-adjusted total kidney volume, the rate of kidney growth reduced significantly from 7.33% to 0.66% annually, since commencement of the tolvaptan therapy. The liver enzyme profile and serum sodium level and osmolality were constantly within normal ranges. In Korea, this is the first reported case of a patient with ADPKD who received tolvaptan treatment for more than 1 year. This case demonstrates that long-term tolvaptan treatment appears to be safe, well tolerated, and effective for ADPKD.

A Subgroup IB Isolate of Cucumber mosaic virus Isolated from Lagenaria leucantha var. gourda (표주박(Lagenaria leucantha var. gourda)에서 분리한 서브그룹 IB계통의 Cucumber mosaic virus)

  • Oh, Sun-Mi;Hong, Jin-Sung;Ryu, Ki-Hyun;Lee, Gung-Pyo;Choi, Jang-Kyung
    • Research in Plant Disease
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    • v.15 no.3
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    • pp.254-258
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    • 2009
  • An isolate of Cucumber mosaic virus (CMV), called as Lag-CMV, was identified from Lagenaria leucantha var. gourda showing mosaic symptom, and its properties was compared to Fny-CMV (subgroup IA) and As-CMV (subgroup IB) by host reaction in several indicator plants, dsRNA analysis, RT-PCR analysis, restriction enzyme profile of the PCR products and nucleotide sequence of coat protein gene. Lag-CMV was similar to As-CMV used as a control CMV by the induced chlorotic spot on inoculated leaves and mosaic symptoms on upper leaves of N. tabacum. cv. Xanthi nc. In the cucumber and zucchini squash, Lag-CMV and As-CMV induced a mild mosaic symptoms than that of Fny-CMV. Size and shapes of local lesions on Chenophodium amaranticolor and Vigna unguiculata induced by Lag-CMV was similar those by Fny-CMV or As-CMV. In experiments of dsRNA profiles and RT-PCR analysis of coat protein gene, Lag-CMV was come within subgroup I CMV. Moreover, restriction enzyme analysis using EcoRI, SalI, MspI, XhoI, and HindIII of the RTPCR products and nucleotide sequence analysis of the coat protein gene showed that Lag-CMV belong to a member of CMV subgroup IB of the same to As-CMV.

Small CNN-RNN Engraft Model Study for Sequence Pattern Extraction in Protein Function Prediction Problems

  • Lee, Jeung Min;Lee, Hyun
    • Journal of the Korea Society of Computer and Information
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    • v.27 no.8
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    • pp.49-59
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    • 2022
  • In this paper, we designed a new enzyme function prediction model PSCREM based on a study that compared and evaluated CNN and LSTM/GRU models, which are the most widely used deep learning models in the field of predicting functions and structures using protein sequences in 2020, under the same conditions. Sequence evolution information was used to preserve detailed patterns which would miss in CNN convolution, and the relationship information between amino acids with functional significance was extracted through overlapping RNNs. It was referenced to feature map production. The RNN family of algorithms used in small CNN-RNN models are LSTM algorithms and GRU algorithms, which are usually stacked two to three times over 100 units, but in this paper, small RNNs consisting of 10 and 20 units are overlapped. The model used the PSSM profile, which is transformed from protein sequence data. The experiment proved 86.4% the performance for the problem of predicting the main classes of enzyme number, and it was confirmed that the performance was 84.4% accurate up to the sub-sub classes of enzyme number. Thus, PSCREM better identifies unique patterns related to protein function through overlapped RNN, and Overlapped RNN is proposed as a novel methodology for protein function and structure prediction extraction.

Association of Cytochrome-17 (MspA1) Gene Polymorphism with Risk of Gall Bladder Stones and Cancer in North India

  • Dwivedi, Shipra;Agrawal, Sarita;Singh, Shraddha;Madeshiya, Amit Kumar;Singh, Devendra;Mahdi, Abbas Ali;Chandra, Abhjeet
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.13
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    • pp.5557-5563
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    • 2015
  • Background: Cholelithiasis is associated in 54%-98% of patients with carcinoma of the gallbladder, and a high incidence among females suggests a role of female hormones in the etiology of the disease. Cytochrome $P450C17{\alpha}$ (CYP-17) is a key enzyme involved in estrogen metabolism and polymorphisms in CYP-17 are associated with altered serum levels of estrogens. Thus, we investigated whether the CYP-17 MspA1 gene polymorphism might impact on risk of gall bladder cancers or gallstones, as well as to determine if this gene polymorphism might be linked with estrogen serum levels and lipid profile among the North Indian gall bladder cancer or gallstone patients. Materials and Methods: CYP-17 gene polymorphisms (MspA1) were genotyped with PCR-RFLP in cancer patients (n=96), stone patients (n=102), cancer + stone patients (n=52) and age/sex matched control subjects (n= 256). Lipid profile was estimated using a commercial kit and serum estrogen was measured using ELISA. Results: The majority of the patients in all groups were females. The lipid profile and estrogen level were significantly higher among the study as compared to control groups. The frequency of mutant allele A2 of CYP17 MspA1 gene polymorphism was higher among cancer (OR=5.13, 95% CI+3.10-8.51, p=0.0001), stone (OR=5.69, 95%CI=3.46-9.37, p=0.0001) and cancer + stone (OR=3.54, 95%CI=1.90-6.60, p=0.0001) when compared with the control group. However there was no significant association between genotypes of CYP17 MspA1 gene polymorphism and circulating serum level of estrogen and lipid profile. Conclusions: A higher frequency of mutant genotype A1A2 as well as mutant allele A2 of CYP-17 gene polymorphism is significantly associated with risk of gallbladder cancer and stones. Elevated levels of estrogen and an altered lipid profile can be used as predictors ofgall bladder stones and cancer in post menopausal females in India.

Proteomic Changes in Odae Polished White Rice Grown at Different Cultivation Conditions (재배환경에 따라 변화하는 오대벼 백미의 단백질체 분석)

  • Lee, Ju-Young;Lee, Jin-Woo;Kim, Young-Ran;Yeom, Yu-Jin;Lim, Jin-Kyu
    • Journal of Applied Biological Chemistry
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    • v.55 no.2
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    • pp.79-83
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    • 2012
  • It has been known that the proteome profiles in the period of growth and development of rice are changed by the growth conditions including temperature, soil, and fertilization. In this study, the proteome profiles of Odae polished white rice grown in Chulwon and Chilgog were compared on 2-dimensional(D) gels. The differentially expressed proteins were selected from the 112 identified total proteins and classified into functional groups. The most significantly differentially expressed proteins were stress responsive proteins; Ent-kaur-16-ene synthase, which is responsible for synthesizing a plant hormone gibberellin, was expressed in Chulwon rice and heat shock proteins were in Chilgog rice, respectively. Xylanase inhibitor protein, which inhibits the enzyme xylanase produced by pathogenic fungi and Bacilli, was expressed significantly high in Chilgog rice grown at high temperature. Differential expressions of transporter proteins were observed both in Chulwon and Chilgog rice. Regarding the facts that Chilgog rice contained relatively higher amount of proteins than Chulwon rice and Chulwon rice showed large number of proteins were differentially expressed, it can be concluded that different cultivation conditions could change the protein expression profiles in rice in various ways, including elevation of protein amount or differential expressions of specific proteins, etc. The results suggest that the characteristics of the profiles of the proteome in the polished white rice are definitely changed by the environmental factors including high temperature. The results can be utilized for the development of the proper cultivation conditions for the production of high quality rice with good palatability.

Studies on the Immobilization of Enzymes and Microoganism Part 1. Immobilizing Method of Glucose Oxidase by Gamma Radiation (효소 및 미생물의 고정화에 관한 연구 제1보. 방사선조사에 의한 Glucose Oxidase의 고정화법)

  • Kim, Sung-Kih
    • Microbiology and Biotechnology Letters
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    • v.7 no.1
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    • pp.1-8
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    • 1979
  • A new method for immobilization of glucose oxidate by the aerobic gamma radiation of synthetic monomers was developed. The radiocopolymerization was conducted aerobically at -70 to -8$0^{\circ}C$ with the mixture of several polyfunctional esters, acrylates and native enzyme. The retained activity of immobilized glucose oxidase was about 50 to 55% when a NK 23G ester, acrylamide-bis and water mixture (1:1:2) in cold toluene treated with 450 krad of gam-ma radiation. The radiation dose did not influence significantly to the enzyme activity. The solvents used to prepare the beads of glucose oxidase and monomers were toluene, n-hexane, petoleum ether and chloroform. 0.05M tris-gycerol (pH 7.0) was a more suitable bugger solution for immobilizing the enzyme than was 0.02M phosphate. Immobilization of glucose oxidase shifted the optimum pH for its reaction from 6.0 to 6.5. The pH profile for the immobilized enzyme showed a broad range of optimum activity while the native enzyme gave a sharp pick for its optimum pH value. The immobilized enzyme reaction temperature was at the range of 30~4$0^{\circ}C$.

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A Study of the Diversity and Profile for Extracellular Enzyme Production of Aerobically Cultured Bacteria in the Gut of Muraenesox cinereus (갯장어(Muraenesox cinereus) 장으로부터 호기적 조건에서 분리된 미생물의 다양성 및 세포외 효소 생산능 분석에 관한 연구)

  • Lee, Yong-Jik;Oh, Do-Kyoung;Kim, Hye Won;Nam, Gae-Won;Sohn, Jae Hak;Lee, Han-Seung;Shin, Kee-Sun;Lee, Sang-Jae
    • Journal of Life Science
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    • v.29 no.2
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    • pp.248-255
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    • 2019
  • This research confirmed the diversity and characterization of gut microorganisms isolated from the intestinal organs of Muraenesox cinereus, collected on the Samcheonpo Coast and Seocheon Coast in South Korea. To isolate strains, Marine agar medium was basically used and cultivated at $37^{\circ}C$ and pH7 for several days aerobically. After single colony isolation, totally 49 pure single-colonies were isolated and phylogenetic analysis was carried out based on the result of 16S rRNA gene DNA sequencing, indicating that isolated strains were divided into 3 phyla, 13 families, 15 genera, 34 species and 49 strains. Proteobacteria phylum, the main phyletic group, comprised 83.7% with 8 families, 8 genera and 26 species of Aeromonadaceae, Pseudoalteromonadaceae, Shewanellaceae, Enterobacteriaceae, Morganellaceae, Moraxellaceae, Pseudomonadaceae, and Vibrionaceae. To confirm whether isolated strain can produce industrially useful enzyme or not, amylase, lipase, and protease enzyme assays were performed individually, showing that 39 strains possessed at least one enzyme activity. Especially the Aeromonas sp. strains showed all enzyme activity tested. This result indicated that isolated strains have shown the possibility of the industrial application. Therefore, this study has contributed for securing domestic genetic resources and the expansion of scientific knowledge of the gut microbial community in Muraenesox cinereus of South Korea.

In Situ PCR on the Glass Slide Using the Conventional DNA Thermal Cycler (기존의 핵산증폭기를 사용한 유리슬라이드상에서의 원위치 중합효소 연쇄반응)

  • 오정균;장진수;이재영
    • Korean Journal of Microbiology
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    • v.39 no.3
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    • pp.197-200
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    • 2003
  • In order to establish effective in situ PCR on the glass slide using the conventional DNA thermal cycler, several parameters should be considered. These include full accessibility of PCR reagents into the cells, prevention of diffusing PCR products out of the cells, loss of PCR reagents by nonspecific adherence onto the glass slide, dryness of PCR reagents by heat, and heat conductivity from the heat block to the glass slide. Especially, to guarantee the full accessibility of PCR reagents to sample, relatively higher concentration of PCR reagents (particularly 4.5 mM of $Mg^{++}$) was required while 5 to 10 units/50 ${\mu}l$ reaction of Taq enzyme was enough as long as the step of pre-PCR incubation was included. Dryness of sample was prevented by addition of distilled water into the empty slots in the heat block, thereby providing the reproducible temperature-time profile of PCR. Observed temperature was lower than the programmed temperature by 3 to $4^{\circ}C$.