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A Bacteriocin of 5-kDa in Size Secreted by Bacillus subtilis 168 (Bacillus subtilis 168 균주가 분비하는 5 kDa 크기의 Bacteriocin)

  • Kwon, Gun-Hee;Lee, Hwang-A;Kim, Jeong-Hwan
    • Microbiology and Biotechnology Letters
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    • v.38 no.2
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    • pp.163-167
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    • 2010
  • Bacillus subtilis 168 secreted antimicrobial substance(s) into culture medium and culture supernatant inhibited growth of some Gram positive bacteria. B. cereus and Listeria monocytogenes were the most sensitive organisms. The antimicrobial activity was destroyed when culture supernatant was treated by protease and proteinase K, indicating the proteinous nature of the substance (bacteriocin). The molecular weight of the bacteriocin was estimated to be 5 kDa by Tricine SDS-PAGE. B. cereus ATCC 14579 cells were killed when exposed to the bacteriocin, indicating that the mode of inhibition was bacteriocidal. These results show that B. subtilis 168 could be useful as a starter for fermented foods such as cheonggukjang where B. cereus contamination is a major concern.

Decolorization of dyes by a purified laccase from Coprinus comatus (정제된 먹물버섯(Coprinus comatus) laccase에 의한 염료 탈색)

  • Kim, Su Yeon;Choi, Ji Young;Choi, Hyoung T.
    • Korean Journal of Microbiology
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    • v.53 no.2
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    • pp.79-82
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    • 2017
  • An inky cap, Coprinus comatus synthesizes and secretes a laccase in the liquid yeast extract peptone dextrose medium. We have successfully purified the enzyme through the ion-exchange chromatography and the preparative gel electrophoresis. The estimated molecular weight was 67 kDa by the SDS-PAGE analysis. Optimum pH was pH 4.3 and optimum temperature was $25^{\circ}C$. The Km value was 0.45 mM and the Vmax was 0.0132 OD/min/unit for o-tolidine. Purified laccase showed 49.3% decolorizing activity against remazol brilliant blue R (RBBR) and 41.6% decolorizing activity against Poly R-478 after 12 h incubation.

Purification and biochemical characterization of two novel antigens from Leishmania major promastigotes

  • Zeinali, Majid;Ardestani, Sussan K.;Kariminia, Amina
    • Parasites, Hosts and Diseases
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    • v.45 no.4
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    • pp.287-293
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    • 2007
  • The identification and characterization of antigens that elicit human T cell responses is an important step toward understanding of Leishmania major infection and ultimately in the development of a vaccine. Micropreparative SDS-PAGE followed by electro transfer to a PVDF membrane and elution of proteins from the PVDF, was used to separate 2 novel proteins from L. major promastigotes, which can induce antibodies of the IgG2a isotype in mice and also are recognized by antisera of recovered human cutaneous leishmaniasis subjects. Fractionation of the crude extract of L. major revealed that all detectable proteins of interest were present within the soluble Leishmania antigens (SLA). Quantitation of these proteins showed that their expression in promastigotes is relatively very low. Considering the molecular weight, immunoreactivity, chromatographic and electrophoretic behavior in reducing and non-reducing conditions, these proteins are probably 2 isoforms of a single protein. A digest of these proteins was resolved on Tricine-SDS-PAGE and immunoreactive fragments were identified by human sera. Two immunoreactive fragments (36.4 and 34.8 kDa) were only generated by endoproteinase Glu-C treatment. These immunoreactive fragments or their parent molecules may be ideal candidates for incorporation in a cocktail vaccine against cutaneous leishmaniasis.

Alterations in Protein and Amino acid Contents During Growth on Culture Cells of Wild Viola (Viola patrinii DC.) (야생 흰 제비꽃(Viola patrinii DC.) 배양세포의 생성과정 중 단백질 및 아미노산의 함량변화)

  • 정용모;임현희;조영수;정정한;이재헌;서정해;권오창
    • Korean Journal of Plant Tissue Culture
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    • v.27 no.3
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    • pp.181-184
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    • 2000
  • Proteins in friable and compact calli of Viola patrinii DC. were analysed. The protein contents in friable calli were lower than those in compact calli. In suspension culture, it increased to the maximum after 3 weeks culture from inoculation and decreased after 4 weeks culture. Several strong lavel signals were detected with the SDS-PAGE analysis. The polypeptides of 28, 31 and 35 KD were observed from the friable cell culture, from the compact cell culture strong band of 30 KD was determined, indicating that these polypeptides may be the major protein occurring during their cultures. Changes in amino acid contents during culture of the viola suspension cells were investigated the amino acid contents were greatest between two and three weeks culture of the viola suspension cells.

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Molecular cloning and foreign gene expression of restriction endonuclease fragments of the Hc nuclear polyhedrosis virus DNA (Hc nuclear polyhedrosis virus DNA 제한효소절편의 molecular cloning 과 외래 유전자 발현)

  • Lee, Keun-Kwang
    • Journal of fish pathology
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    • v.8 no.1
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    • pp.31-36
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    • 1995
  • Hc nuclear polyhedrosis virus DNA genome was digested with EcoRI endonuclease, these partial fragments were recombined into the pUC8 plasmid vector and transformed in E. coli JM 83 cell. The genome DNA has 24 EcoRI fragments and 12 fragments of them were subcloned. The four recombinants were named as eNP-O, eNP-Q, eNP-R and eNP-S. The expression of foregin gene of the recombinants was investigated by analysing protein patterns on the SDS-PAGE. The eNP-O, eNP-Q and eNP-R were expressed a different weight of protein as comparision with potypeptide bands of E. coli JM 83 host cell.

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An Evaluation of Changes in the Allergenicity of Kochujang upon Preparation Using Aloe Extract

  • Son, Bo-Kyung;Huh, Yoon-Ee;Kim, Jung-Yun;Noh, Geon-Woong;Lee, Sang-Sun
    • Nutritional Sciences
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    • v.9 no.4
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    • pp.317-322
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    • 2006
  • Soybeans are well-known as allergenic foods. Koreans consume large amounts of soybean foods, such as kochujang, which have gone through the fermentation process. To lower the allergenicity of these foods, we prepared hypo allergenic kochujang with aloe extract (AK). A sensory evaluation was conducted along with a clinical evaluation that used a double-blind, placebo-controlled food challenge (DBPCFC) test These tests were designed to evaluate the acceptability of the fermented foods. In comparison to normal kochujang (NK), AK elicited a higher sensory test score, and the rate of positive reactions in atopic dermatitis patients during the DBPCFC test was reduced. Methods of protein extraction, protein quantitation with sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE), and protein identification using two-dimensional (2D) gel electrophoresis were performed for both NK and AK to compare the functional factors. We found a reduction in the levels of high molecular proteins even though the bands of the proteins had not entirely disappeared, indicating that the boiling and fermentation process changed the soybean protein patterns. The rate of the reduction of high molecular proteins was more effective in the AK. In conclusion, AK can be recognized as a food with hypoallergenic effect.

Studies on the Isolation of Phycobiliprotein from S. platensis (S. platensis로부터 phycobiliprotein의 분리에 관한 연구)

  • Kim Jum-Ji;Kim Yoon-Kyoun;Lee Mi-Young
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.5 no.5
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    • pp.484-489
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    • 2004
  • Isolation of phycobiliprotein from S. platensis was performed by using $30-60{\%}$ ammomium sulfate fractionation, Sephadex G-100 gel filtration and DEAE-Sephacel anionic exchange chromatography. Isolated phycobiliprotein was determined to be a c-phycocyanin with a maximum absorption wavelength at 620 nm. This phycobiliprotein consisted of $({\alpha}$ and $({\beta}$ subunit when analyzed through SDS-PAGE. The molecular weights of $({\alpha}$ and $({\beta}$ subunit were 14.5 kDa and 16 kDa, respectively. The native molecular weight of phycobiliprotein through gel filtration was about 100 kDa. These results show that the structure of phycobiliprotein from S. platensis might be aggregated form of $({\alpha}{\beta})_{3}-trimer$.

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Cloning and Characterization of Cellulase Gene (cel5B) from Cow Rumen Metagenome

  • Kang, Tae-Ho;Kim, Min-Keun;Barman, Dhirendra Nath;Kim, Jung-Ho;Kim, Hoon;Yun, Han-Dae
    • Journal of agriculture & life science
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    • v.46 no.2
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    • pp.129-137
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    • 2012
  • A carboxymethyl cellulase gene, cel5B, was cloned, sequenced, and expressed in Escherichia coli. pRCS20 in E. coli was identified from metagenomic cosmid library of cow rumen for cellulase activity on a carboxymethyl cellulose agar plates. Cosmid clone (RCS20) was partially digested with Sau3AI, ligated into BamHI site of pBluescript II SK+ vector, and transformed into E. coli $DH5{\alpha}$. The insert DNA of 1.3 kb was obtained, designated cel5B, which has the activity of hydrolyzation of CMC. The cel5B gene had an open reading frame (ORF) of 1,059 bp encoding 352 amino acids with a signal peptide of 48 amino acids and the conserved region, VIYEIYNEPL, belongs to the glycosyl hydrolase family 5. The molecular mass of Cel5B protein expressed from E. coli $DH5{\alpha}$ exhibited to be about 34 kDa by CMC-SDS-PAGE. The optimal pH was 8.0, and the optimal temperature was about $50^{\circ}C$ for its enzymatic activity.

Studies on standardization and characterization of recombinant interferon alia

  • Kim, Gi-Hyun;Shin , Won;Jung , Ja-Young;Park, Young-Ju;Joung , Jee-Won;Oh, Il-Ung;Jin, Jae-Ho;Kim, Seo-Mi;Jung , Sang-Mi
    • Proceedings of the PSK Conference
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    • 2002.10a
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    • pp.329.2-330
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    • 2002
  • This study was intended to establish test methods equivalent to those of "Interferon alfa-2 concentrated solution" monograph in European Pharmacophoeia(EP). Two recombinant interferon alfa concentrated solutions manufactured in Korea were tested according to the monograph of EP. Tests of identification(biological activity. isoelectric focusing. SDS-PAGE under reducing condition, peptide mapping). related proteins. impurities of moducular masses differing from that of interferon alfa-2(SDS-PAGE under reducing and non-reducing condition). bacterial endotoxin, protein, potency, host-cell-derrved proteins. and host-cell-derived DNA were performed in the laboratories of manufacrues and division of biotechnology. KFDA. The results of this study showed that specitications of interfenon alfa concentrated solutions manufactured in Korea were within the aceptance criteria of EP. Based on the study. specitications and test methods for interferon alfa concentrated solution can be established according to the monograph of EP suggesting the revision of Minimum requirements for biological products

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Quality properties of whole milk powder on chicken breast emulsion-type sausage

  • Kang, Kyu-Min;Lee, Sol-Hee;Kim, Hack-Youn
    • Journal of Animal Science and Technology
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    • v.63 no.2
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    • pp.405-416
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    • 2021
  • The aim of the study was to determine the effect of whole milk powder (WMP) as heterologous proteins on chicken breast emulsion-type sausages. The quality properties of WMP on such chicken breast emulsion-type sausages were investigated by measuring the proximate composition, pH, color, cooking yield, protein solubility, and by applying other methods, such as texture profile analysis (TPA), microphotograph, sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and electronic nose. The crude fat, protein, and ash contents of 15% WMP samples were significantly higher than the control samples (p < 0.05). The redness of the cooked samples significantly increased with an increase in the WMP contents (p < 0.05). The cooking yield of WMP treated samples was significantly higher than the control sample (p < 0.05). Additionally, the hardness, gumminess, and chewiness of WMP treated samples were significantly higher than the control sample (p < 0.05). The sarcoplasmic and myofibrillar proteins of samples containing 15% WMP were significantly higher than the control samples (p < 0.05). The result of SDS-PAGE showed that the C protein, sarcoplasmic protein, actin, and tropomyosin increased with an increase in the WMP contents. The principal component analysis plot of WMP-treated samples was clearly different from that of the control samples. Based on these results, it was predicted that WMP could be useful as heterologous protein on emulsion-type sausage.