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Identification of LAB and Fungi in Laru, a Fermentation Starter, by PCR-DGGE, SDS-PAGE, and MALDI-TOF MS

  • Ahmadsah, Lenny S.F.;Kim, Eiseul;Jung, Youn-Sik;Kim, Hae-Yeong
    • Journal of Microbiology and Biotechnology
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    • v.28 no.1
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    • pp.32-39
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    • 2018
  • Samples of Laru (a fermentation starter) obtained from the upper part of Borneo Island were analyzed for their lactic acid bacteria (LAB) and fungal diversity using both a culture-independent method (PCR-DGGE) and culture-dependent methods (SDS-PAGE and MALDI-TOF MS). Pediococcus pentosaceus, Lactobacillus brevis, Saccharomycopsis fibuligera, Hyphopichia burtonii, and Kodamaea ohmeri were detected by all three methods. In addition, Weissella cibaria, Weissella paramesenteroides, Leuconostoc citreum, Leuconostoc mesenteroides, Lactococcus lactis, Rhizopus oryzae/Amylomyces rouxii, Mucor indicus, and Candida intermedia were detected by PCR-DGGE. In contrast, Lactobacillus fermentum, Lactobacillus plantarum, Pichia anomala, Candida parapsilosis, and Candida orthopsilosis were detected only by the culture-dependent methods. Our results indicate that the culture-independent method can be used to determine whether multiple laru samples originated from the same manufacturing region; however, using the culture-independent and the two culture-dependent approaches in combination provides a more comprehensive overview of the laru microbiota.

A Study on the Pattern of Proteins on Electrophoresis in the Porcine Follicular Fluid During Atresia (폐쇄에 따른 돼지 여포액내 단백질의 전기영동 양상에 관한 연구)

  • Kim, Jong-Heup;Yoon, Yong-Dal;Kim, Moon-Kyoo
    • Clinical and Experimental Reproductive Medicine
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    • v.13 no.1
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    • pp.39-51
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    • 1986
  • This experiment has been done to evaluate the relationship between the follicular atresia and the protein patterns on electrophoreais of the follicular fluids in porcine ovary. The protein concentration of the follicular fluids was lower than that of serum, and gradually decreased as the follicle siae became larger. The number of protein bands of follicular fluid on electrophoresis was less than that of serum, and gradually increased as follicle size became larger. Three specific bands were detected on disc PAGE and one band(M W. 75,000) on SDS PAGE in the follicular fluids, while not in serum. One band (A) at ${\beta}$-globulin region on disc PAGE became heavier, as follicles became atretic. Two bands less than(M. W. 20,000) were detected only in the large follicular fluid. Another band(M. W. 43,000) was not detected in necrotic group, whereas all other groups showed it. It could be concluded that the component and composition of the proteins follicular fluids changes according to the follicular size during atresia. Therefore detection of the changing pattern of proteins in the follicular fluid can be used as a basic criterion for the identification of follicular atretic stage.

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Memory Compaction Scheme with Block-Level Buffer for Large Flash Memory

  • Chung, Weon-Il;Li, Liangbo
    • International Journal of Contents
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    • v.6 no.4
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    • pp.22-29
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    • 2010
  • In flash memory, many previous garbage collection methods only merge blocks statically and do not consider the contents of buffer. These schemes may cause more unnecessary block erase operations and page copy operations. However, since flash memory has the limitation of maximum rate and life cycle to delete each block, an efficient garbage collection method to evenly wear out the flash memory region is needed. This paper proposes a memory compaction scheme based on block-level buffer for flash memory. The proposed scheme not only merges the data blocks and the corresponding log block, but also searches for the block-level buffer to find the corresponding buffer blocks. Consequently, unnecessary potential page copying operations and block erasure operations could be reduced, thereby improving the performance of flash memory and prolonging the lifetime of flash memory.

$Ca^{2+}$ is Required to Make Functional Malate Synthase in Corynebacterium glutamicum

  • Kim, Hyung-Joon;Kim, Jae-Ho;Lee, Heung-Shick
    • Journal of Microbiology and Biotechnology
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    • v.7 no.6
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    • pp.435-437
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    • 1997
  • The role of $Ca^{2+}$ in making functional malate synthase in Corynebacterium glutamicum was investigated using the cloned DNA coding for the enzyme. Introduction of cloned aceB into C. glutamicum overexpressed malate synthase as judged by SDS-PAGE. However, the increase in enzyme activity of the expressed malate synthase did not match the level of overexpression observed in SDS-PAGE. Addition of $Ca^{2+}$ to the growth medium specifically increased the activity. The malate synthase could be stained with ruthenium red in a $Ca^{2+}$-specific manner. This agrees with the previous observation which reported a potential $Ca^{2+}$-binding domain in the N-terminal region of the protein.

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Chemical and Sensory Properties of New Gluten-free Food Products: Rice and Corn Tarhana

  • Yalcin, Erkan;Celik, Sueda;Koksel, Hamit
    • Food Science and Biotechnology
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    • v.17 no.4
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    • pp.728-733
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    • 2008
  • New gluten-free food product (tarhana) was produced using rice and com flours. Chemical and sensory properties of the tarhana samples were investigated and compared with those of traditional wheat tarhana. Generally, sensory analysis results indicated that utilization of com and rice flours in tarhana resulted in acceptable soup properties in terms of some of the sensory properties. The changes in electrophoretic properties of the proteins of the tarhana samples were also studied during the tarhana production. Sodium dodecyl sulfate-polyacryamide gel electrophoresis (SDS-PAGE) results showed that relative intensities of some of the protein bands in tarhana samples generally decreased during fermentation. The decrease was more obvious at the larger molecular weight region. Corn and rice tarhana seem to be promising food products for the celiac patients who have limited choice of cereal based foods.

A Web Surfing Assistant for Improved Web Accessibility (웹 접근성 향상을 위한 웹 서핑 도우미)

  • Lee SooCheol;Lee Sieun;Hwang Eenjun
    • Journal of KIISE:Software and Applications
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    • v.31 no.9
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    • pp.1180-1195
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    • 2004
  • Due to the exponential increase of information, search and access for the Web information or service takes much time. Web information is represented through several web pages using hyperlinks and each web page is contains several topics. However. most existing web tools don't reflect such web authoring tendencies and treat it as an independent information unit. This inconsistency yields inherent problems in web browsing and searching. In this paper, we propose a web surfing assistant called LinkBroker that provides collodion pages. They are composed of relevant information extracted from several web pages that have table and frame structure in order to improve accessibility to web information. Especially, the system extracts a set of web pages that are logically connected and groups those pages using table and frame tags. Then, essential information blocks in each page of a group are extracted to construct an integrated summary page. It Provides a comprehensive view to user and one cut way to access distributed information. Experimental results show the effectiveness and usefulness of LinkBroker system.

Document Image Layout Analysis Using Image Filters and Constrained Conditions (이미지 필터와 제한조건을 이용한 문서영상 구조분석)

  • Jang, Dae-Geun;Hwang, Chan-Sik
    • The KIPS Transactions:PartB
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    • v.9B no.3
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    • pp.311-318
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    • 2002
  • Document image layout analysis contains the process to segment document image into detailed regions and the process to classify the segmented regions into text, picture, table or etc. In the region classification process, the size of a region, the density of black pixels, and the complexity of pixel distribution are the bases of region classification. But in case of picture, the ranges of these bases are so wide that it's difficult to decide the classification threshold between picture and others. As a result, the picture has a higher region classification error than others. In this paper, we propose document image layout analysis method which has a better performance for the picture and text region classification than that of previous methods including commercial softwares. In the picture and text region classification, median filter is used in order to reduce the influence of the size of a region, the density of black pixels, and the complexity of pixel distribution. Futhermore the classification error is corrected by the use of region expanding filter and constrained conditions.

Detection of Mitotic Centromere-Associated Kinesin (MCAK) During Cell-Cycle Progression of Human Jurkat T Cells Using Polyclonal Antibody Raised Against Its N- Terminal Region Overexpressed in E. coli

  • Jun, Do-Youn;Rue, Seok-Woo;Kim, Byung-Woo;Kim, Young-Ho
    • Journal of Microbiology and Biotechnology
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    • v.13 no.6
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    • pp.912-918
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    • 2003
  • Mitotic centromere-associated kinesin (MCAK), which is a novel kinesin with a central motor domain, is believed to playa role in mitotic segregation of chromosome during the M phase of the cell cycle. In the present study, it is shown that a rabbit polyclonal antibody has been produced using the N-terminal region (187 aa) of human MCAK expressed in E. coli as the antigen. To express the N-terminal region in E. coli, the MCAK cDNA fragment encoding N-terminal 187 aa was obtained by PCR and was then inserted into the pET 3d expression vector. Molecular mass of the N-terminal region overexpressed in the presence of IPTG was 23.2 kDa on SDS-PAGE, and the protein was insoluble and mainly localized in the inclusion body that could be easily purified from the other cellular proteins. The N-terminal region was purified by electro-elution from the gel after the inclusion body was resolved on the SDS-PAGE. The antiserum obtained after tertiary immunization with the purified protein specifically recognized HsMCAK when subjected to Western blot analysis, and showed a fluctuation of the protein level during the cell cycle of human Jurkat T cells. Synchronization of the cell-cycle progression required for recovery of cells at a specific stage of the cell cycle was performed by either hydroxyurea or nocadazole, and subsequent release from each blocking at 2, 4, and 7 h. Northern and Western analyses revealed that both mRNA and protein of HsMCAK reached a maximum level in the S phase and declined to a basal level in the G1 phase. These results indicate that a polyclonal antibody raised against the N-terminal region (187 aa) of HsMCAK, overexpressed in E. coli, specifically detects HsMCAK (81 kDa), and it can analyze the differential expression of HsMCAK protein during the cell cycle.

Characterization of PpsR, a Transcriptional Repressor of the Expression of Photosysem Gene, from Rhodobacter sphaeroides

  • Cho, Seung-Hyun;Kang, Sa-Ouk
    • Proceedings of the Korean Biophysical Society Conference
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    • 2002.06b
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    • pp.51-51
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    • 2002
  • PpsR from the facultative photohetrotroph Rhodobacter sphaeroides is involved in repression of photo system gene expression. SDS-PAGE analysis showed that some portion of PpsR is oxidized so that intra- or inter-disulfide bond is formed between the two cysteins in each subunit. The disulfide bond was reduced by dithiothreitol and the binding activity to puc promoter region was increased.(omitted)

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Difference in Susceptibility of Tyrosine Residue to Oxidative Iodination between a Thioredoxin Box Region and a Hormonogenic Region

  • Sok, Dai-Eun;Charles J.Sih
    • Archives of Pharmacal Research
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    • v.24 no.5
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    • pp.446-454
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    • 2001
  • Peptide fragments, isolated from proteolytic cleavage of thyroglobulin at specific sites, were examined for the iodination of tyrosine residues. The 50 kDa polypeptide, which was prepared from digestion of bovine thyroglobulin and continuous preparative SDS-PAGE, was subjected to reduction with DTT and alkylation with iodoacetic acid to generate S-car-boxymethylated peptide derivative, which was further hydrohysed by endoproteinase-Asp-N. Peptide products were separated by RP-HPLC, and each fraction was analyzed by LC/ESI-MS and MALDI-MS analyses. Based on the specificity of endoproteinase-Asp-N andthe mass spectra data, a peptide fragment turned out to correspond to a peptide, DALCCVKCPEGSYFQ (1438-1452), characterized by the presence of a thioredoxin box (CVKC) and a tyrosine residue. In addition, another peptide fragment (1453-1465) containing a thioredoxin box (CIPC) and a tyrosine residue was also observed. However, any evidence of iodination of the tyrosine residue present in these peptides was not provided. Meanwhile, tyrosine residues in the peptides, DVEEALAGKYLAGRFA (1366-1381) and DYSGLLLAFQVFLL (1290-1303) were found to be iodinated; mono- or diiodinated tyrosine residues, characteristic of a hormogenic site, existed in both peptides. In addition, the tyrosine residue in the peptide (1218-1252), corresponding to a hormonogenic site was also iodinated. Thus, there was a sharp difference of the susceptibility to oxidative iodination between the tyrosine residue in a hormonogenic site and that in a thioredoxin region. From these results, it is suggested that polypeptide region adjacent to tyrosine residues may govern the susceptibility of tyrosine to oxidative iodination.

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