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Arabinoxylo- and Arabino-Oligosaccharides-Specific α-ʟ-Arabinofuranosidase GH51 Isozymes from the Amylolytic Yeast Saccharomycopsis fibuligera

  • Park, Tae Hyeon;Choi, Chang-Yun;Kim, Hyeon Jin;Song, Jeong-Rok;Park, Damee;Kang, Hyun Ah;Kim, Tae-Jip
    • Journal of Microbiology and Biotechnology
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    • v.31 no.2
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    • pp.272-279
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    • 2021
  • Two genes encoding probable α-ʟ-arabinofuranosidase (E.C. 3.2.1.55) isozymes (ABFs) with 92.3% amino acid sequence identity, ABF51A and ABF51B, were found from chromosomes 3 and 5 of Saccharomycopsis fibuligera KJJ81, an amylolytic yeast isolated from Korean wheat-based nuruk, respectively. Each open reading frame consists of 1,551 nucleotides and encodes a protein of 517 amino acids with the molecular mass of approximately 59 kDa. These isozymes share approximately 49% amino acid sequence identity with eukaryotic ABFs from filamentous fungi. The corresponding genes were cloned, functionally expressed, and purified from Escherichia coli. SfABF51A and SfABF51B showed the highest activities on p-nitrophenyl arabinofuranoside at 40~45℃ and pH 7.0 in sodium phosphate buffer and at 50℃ and pH 6.0 in sodium acetate buffer, respectively. These exoacting enzymes belonging to the glycoside hydrolase (GH) family 51 could hydrolyze arabinoxylo-oligosaccharides (AXOS) and arabino-oligosaccharides (AOS) to produce only ʟ-arabinose, whereas they could hardly degrade any polymeric substrates including arabinans and arabinoxylans. The detailed product analyses revealed that both SfABF51 isozymes can catalyze the versatile hydrolysis of α-(1,2)- and α-(1,3)-ʟ-arabinofuranosidic linkages of AXOS, and α-(1,2)-, α-(1,3)-, and α-(1,5)-linkages of linear and branched AOS. On the contrary, they have much lower activity against the α-(1,2)- and α-(1,3)-double-substituted substrates than the single-substituted ones. These hydrolases could potentially play important roles in the degradation and utilization of hemicellulosic biomass by S. fibuligera.

Construction of a Novel Shuttle Vector for Tetragenococcus species based on a Cryptic Plasmid from Tetragenococcus halophilus

  • Min Jae Kim;Tae Jin Kim;Yun Ji Kang;Ji Yeon Yoo;Jeong Hwan Kim
    • Journal of Microbiology and Biotechnology
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    • v.33 no.2
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    • pp.211-218
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    • 2023
  • A cryptic plasmid (pTH32) was characterized from Tetragenococcus halophilus 32, an isolate from jeotgal, Korean traditional fermented seafood. pTH32 is 3,198 bp in size with G+C content of 35.84%, and contains 4 open reading frames (ORFs). orf1 and orf2 are 456 bp and 273 bp in size, respectively, and their translation products showed 65.16% and 69.35% similarities with RepB family plasmid replication initiators, respectively, suggesting the rolling-circle replication (RCR) mode of pTH32. orf3 and orf4 encodes putative hypothetical protein of 186 and 76 amino acids, respectively. A novel Tetragenococcus-Escherichia coli shuttle vector, pMJ32E (7.3 kb, Emr), was constructed by ligation of pTH32 with pBluescript II KS(+) and an erythromycin resistance gene (ErmC). pMJ32E successfully replicated in Enterococcus faecalis 29212 and T. halophilus 31 but not in other LAB species. A pepA gene, encoding aminopeptidase A (PepA) from T. halophilus CY54, was successfully expressed in T. halophilus 31 using pMJ32E. The transformant (TF) showed higher PepA activity (49.8 U/mg protein) than T. halophilus 31 cell (control). When T. halophilus 31 TF was subculturd in MRS broth without antibiotic at 48 h intervals, 53.8% of cells retained pMJ32E after 96 h, and only 2.4% of cells retained pMJ32E after 14 days, supporting the RCR mode of pTH32. pMJ32E could be useful for the genetic engineering of Tetragenococcus and Enterococcus species.

The Dramatization of Habitus: A Bourdieun Reading of Pygmalion

  • Hwang, Hoon-Sung
    • Journal of English Language & Literature
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    • v.55 no.3
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    • pp.383-398
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    • 2009
  • Based on the Greek myth of Pygmalion and the fairy tale of Cinderella, Shaw's Pygmalion demonstrates a masterful coalescence of these two narrative motifs into a coherent plot scheme. Even more significant is his keen insight into the conflicts created at the tripartite intersection of human activity concerning language/class/culture, which, as the leitmotif, revolves around lessons in language learning. This play basically deals with human transformation and by its very nature, Higgins's experimentation with transforming Eliza cannot stop at language alone. Her cultural transformation ripples over into the realms of gesture and even a unique way of living (modus vivendi) intimately associated with taste and manners, which Bourdieu terms as habitus. By acquiring a new fashion and language, Eliza is reborn as a new lady aspiring to be filled with a newly acquired habitus. While separating her from her old Cockney style, Higgins inculcates Queen's English in Eliza, in which process her changed speech styles gradually transforms and restructures her deportment and manners, finally generating new practices, perceptions and attitudes. The gist of Pygmalion is however less Eliza's ascent into the middle class than her battle for symbolic capital waged at the level of language. By problematizing his contemporary practice of habitus conventionalized and warped by class distinctions based on economic, social and cultural capitals, Shaw creates a new humanist model of man founded on spiritual and rational virtues. In conclusion, Eliza is not a frigid Galatea but a dynamic character that goes through a brilliant transformation of three stages: 1) linguistic; 2) cultural, and 3) humanist. Finally she is built into a "consort battleship" on an equal standing with her sculptor. The process of her character-building cannot be illuminated without resorting to the dynamic notion of habitus, which highlights the process of inculcation, structuring, generation and transposing. Given the overwhelming weight of the heroine's role and the dynamic process of her transformation as the major plot scheme, this play should be christened Galatea in lieu of Pygmalion.

Exploring the molecular characteristics, detoxification functions, and immune responses of two glutathione S-transferases in redlip mullet (Liza haematochelia)

  • Jeongeun Kim;Welivitiye Kankanamge Malithi Omeka;Qiang Wan;Jehee Lee
    • Fisheries and Aquatic Sciences
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    • v.27 no.5
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    • pp.314-328
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    • 2024
  • The mechanism for the elimination of xenobiotics undergoes three different phases of reactions in organisms. Among these, glutathione S-transferases (GSTs) are classified as phase II detoxification enzymes, catalyzing the conjugation of electrophilic substrates to glutathione or reduced hydroperoxides. This study aimed to investigate the molecular characteristics, detoxification functions, and immune responses of GST omega (LhGSTO1) and kappa (LhGSTK1) in redlip mullet. The open reading frames of LhGSTO1 (720 bp) and LhGSTK1 (687 bp) encoded proteins of 239 and 228 amino acids, respectively. Sequence analysis revealed that LhGSTO1 and LhGSTK1 possessed GSH-binding sites in their N-terminal domains. Substrate-binding sites in the C-terminal domain were exclusively identified in LhGSTO1. In the tissue-specific transcription profile analysis, both LhGSTO1 and LhGSTK1 were ubiquitously expressed in all tissues of healthy mullets. Temporal expression analysis of LhGSTO1 and LhGSTK1 in the blood showed that their expression was significantly modulated by polyinosinic:polycytidylic (poly I:C), lipopolysaccharide (LPS), and Lactococcus garvieae. Different chemical and cellular assays were performed to assess the detoxification and cellular protective abilities of the two proteins. A substrate specificity test using the recombinant proteins revealed that both proteins possessed specific activity toward 1-chloro-2,4-dinitrobenzene (CDNB). In the disk diffusion assay, the smallest clearance zones were observed for LhGSTO1 and LGSTK1 against CdCl2. In the cell protection assay, both LhGSTO1 and LhGSTK1 showed significant Cd detoxification ability compared to the control. Collectively, these results demonstrate that GST omega and kappa are involved in host defense against immune stimulants and xenobiotics in redlip mullet.

Development and Evaluation of Sustainable Housing Teaching-Learning Process Plan for Achieving the Global SDGs by Home Economics in Middle School (중학교 가정교과의 SDGs 교육을 위한 지속가능한 주생활 교수·학습 과정안 개발 및 평가)

  • Kim, Eunkyung;Cho, Jaesoon
    • Journal of Korean Home Economics Education Association
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    • v.32 no.2
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    • pp.77-97
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    • 2020
  • The purpose of this study was to develop and evaluate the sustainable housing teaching-learning process plan aimed to achieve the global SDGs through home economics class in middle school that is based on the ADDIE model. The overall objective of the plan was to contribute to cultivating students' sustainable housing values and to creating sustainable lifestyle through everyday practice. The plan consisting of 4 lessons contained various activity and visual resources(4 individual and 4 team activity sheets, 4 reading texts, 1 homework sheet and 1 evaluation sheet, and 7 videos) for students and (4 sets of ppt and 4 reading texts) for teachers. The theme and team activities of each lesson were related to 2~7 targets of 2~3 SDGs, in total 11 targets of 5 SDGs. The plan was implemented to 4 classes of 127 senior students at Y middle school in Cheongju city during the period from the 29th of August to the 18th of September, 2019. The results showed that students were very positive and highly satisfied with not only practical contents but also adequacy of resources and activities of the whole 4-lessons, so that they actively participated in the lessons more than usual and looked forward to learning more about it. They thoroughly enjoyed various team activities such as brain writing, mandal art, visual thinking, making UCC, and planning the sustainable village as well as writing a short reflective journal at the end of each lesson. Students also reported that they highly accomplished the goal of each lesson and the overall objective. It could be concluded that the teaching-learning process plan of 4-lessons could contribute to cultivating students' sustainable housing values and to creating sustainable lifestyle through practicing everyday life. It indicates that home economics is one of the major subjects to contribute to the attainment of global issue of SDGs for OECD education 2030 and to educate the practically acting global citizen.

cDNA Cloning and Stage-Dependant Expression of Arylphorin Gene from Chinese Oak Silkworm, Antheraea pernyi (작잠(Antheraea pernyi) 아릴포린(Arylphorin) 유전자의 cDNA 클로닝 및 아릴포린 유전자의 발육시기 의존성 발현양상)

  • Lee, Sang-Mong;Hwang, Jae-Sam;Park, Nam-Sook;Kim, Yong-Gyun;Kim, Keun-Ki;Son, Hong-Joo;Park, Hyun-Chul;Jin, Byung-Rae
    • Journal of Life Science
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    • v.20 no.8
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    • pp.1193-1200
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    • 2010
  • The cDNA cloning and developmental profiles of the mRNA for A. pernyi arylphorin was determined. The complete A. pernyi arylphorin cDNA sequence comprised 2,234 bp (without the poly $A^+$ tail), including an open reading frame of 2,112 bp beginning with a methionine ATG at bp34. The A. pernyi arylphorin contained 704 amino acids which are highly enriched in aromatic amino acids, phenylalanine and tyrosine. The calculated molecular mass of the A. pernyi arylphorin from the ORF was 83,439 Da. The deduced amino acid sequence of A. pernyi arylphorin showed 78, 71, 62 and 64% identity with those of H. cecropia, M. sexta $\alpha$ subunit, M. sexta $\beta$ subunit and B. mori storage protein. In Northern blot analysis, the A. pernyi arylphorin mRNA only in the fat body of the 5th instar larvae was responsible for gene expression of the protein, and the synthetic activity of the mRNA was detected strongly in the early larvae, but not in the middle or late-stage larvae. In addition, a very weak signal in mRNA activity was detected in pupal stages, but this was considered to be inactive mRNA after reviewing the results of the labeling experiment of this protein.

Practical problem-based teaching·learning process plan to develop and apply to enhance safety awareness in middle school students (중학생의 주생활 안전의식 함양을 위한 실천적 문제 중심 가정과 교수·학습 과정안 개발 및 적용)

  • Song, Eunmi;Cho, Jaesoon
    • Journal of Korean Home Economics Education Association
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    • v.29 no.1
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    • pp.15-33
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    • 2017
  • The purpose of this study was to develop and evaluate a practical problem-based teaching learning process plan for safety in residential environment to raise safety awareness of middle school students. The plan consisting of 4 lessons has been developed and implemented according to the ADDIE model. Various activity materials (26 student's activity sheets and 8 reading texts, and 8 teacher's reading texts) and visual materials (4 sets of pictures & photos and 8 moving pictures) as well as questionnaire were developed for the 4-session lessons. The plans were implemented by the researcher to 4 classes 121 freshmen of M boy's middle school in Kyeongbuk during December 21st to 29th, 2015. Students were highly enjoyed and satisfied with the whole 4-lessons in the aspects such as the level of participation in the lesson, understanding of the contents, adequacy of materials and activities, and usefulness in own's daily life. Students also reported that they were highly aware to practice the contents learned from the lessons in daily family life at home with one's family and recommended to teach the lessons to other schools, too. It can be concluded that the teaching learning process plan for safety in residential environment would raise safety awareness of middle school students through the Home Economics subject.

Development and Application of Practical Problem-based Teaching·Learning Process for Interacting with Neighbors (이웃과 더불어 살아가는 주생활을 위한 실천적 문제 중심 교수·학습 과정안 개발 및 적용)

  • Woo, Yeseul;Cho, Jaesoon
    • Journal of Korean Home Economics Education Association
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    • v.30 no.3
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    • pp.67-90
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    • 2018
  • The purpose of this study was to develop and apply the practical problem-based teaching·learning process plan for 'interacting with neighbors' of home economics subject. The plan consisting of 3 lessons has been developed and implemented according to the ADDIE model. Various activity materials (7 student's activity sheets, 3 reading texts, 1 homework sheet, 3 sets of ppt, 6 videos, and 3 teacher's reading texts) as well as questionnaire were developed for the 3-session lessons. The plans were implemented by the researcher to 204 freshmen, 8 classes, of C middle school in Seoul during september, 2017. The result, of students' lower level of actual participation in interacting with neighbors comparing to their interests in, supported the need of this study. Students were satisfied with the whole 3-lessons in the aspects such as beneficial usage of the contents in their daily life and in building the sense of community, as well as adequacy of materials and activities. Students also reported that they would highly aware to the importance of interacting with neighbors and to practice the contents learned from the lessons in daily life at community. They had an opportunity to reflect one's own attitude to neighbors and recommended to teach it to other schools, too. It can be concluded that the teaching·learning process plan for 'interacting with neighbors' would raise students' housing values living together and attain the overall objective and achievement standards of 2015 home economics middle school curriculum.

Susceptibility of Anthonomus grandis (Cotton Boll Weevil) and Spodoptera frugiperda (Fall Armyworm) to a Cry1Ia-type Toxin from a Brazilian Bacillus thuringiensis Strain

  • Grossi-De-Sa, Maria Fatima;De Magalhaes, Mariana Quezado;Silva, Marilia Santos;Silva, Shirley Margareth.Buffon;Dias, Simoni Campos;Nakasu, Erich Yukio Tempel;Brunetta, Patricia Sanglard Felipe;Oliveira, Gustavo Ramos;De Oliveira Neto, Osmundo Brilhante;De Oliveira, Raquel Sampaio;Soares, Luis Henrique Barros;Ayub, Marco Antonio Zachia;Siqueira, Herbert Alvaro Abreu;Figueira, Edson L.Z.
    • BMB Reports
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    • v.40 no.5
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    • pp.773-782
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    • 2007
  • Different isolates of the soil bacterium Bacillus thuringiensis produce multiple crystal (Cry) proteins toxic to a variety of insects, nematodes and protozoans. These insecticidal Cry toxins are known to be active against specific insect orders, being harmless to mammals, birds, amphibians, and reptiles. Due to these characteristics, genes encoding several Cry toxins have been engineered in order to be expressed by a variety of crop plants to control insectpests. The cotton boll weevil, Anthonomus grandis, and the fall armyworm, Spodoptera frugiperda, are the major economically devastating pests of cotton crop in Brazil, causing severe losses, mainly due to their endophytic habit, which results in damages to the cotton boll and floral bud structures. A cry1Ia-type gene, designated cry1Ia12, was isolated and cloned from the Bt S811 strain. Nucleotide sequencing of the cry1Ia12 gene revealed an open reading frame of 2160 bp, encoding a protein of 719 amino acid residues in length, with a predicted molecular mass of 81 kDa. The amino acid sequence of Cry1Ia12 is 99% identical to the known Cry1Ia proteins and differs from them only in one or two amino acid residues positioned along the three domains involved in the insecticidal activity of the toxin. The recombinant Cry1Ia12 protein, corresponding to the cry1Ia12 gene expressed in Escherichia coli cells, showed moderate toxicity towards first instar larvae of both cotton boll weevil and fall armyworm. The highest concentration of the recombinant Cry1Ia12 tested to achieve the maximum toxicities against cotton boll weevil larvae and fall armyworm larvae were 230 ${\mu}g/mL$ and 5 ${\mu}g/mL$, respectively. The herein demonstrated insecticidal activity of the recombinant Cry1Ia12 toxin against cotton boll weevil and fall armyworm larvae opens promising perspectives for the genetic engineering of cotton crop resistant to both these devastating pests in Brazil.

Cloning and Characterization of Cellulase Gene (cel5C) from Cow Rumen Metagenomic Library (소 반추위 메타게놈에서 새로운 섬유소분해효소 유전자(cel5C) 클로닝 및 유전산물의 특성)

  • Kim, Min-Keun;Barman, Dhirendra Nath;Kang, Tae-Ho;Kim, Jung-Ho;Kim, Hoon;Yun, Han-Dae
    • Journal of Life Science
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    • v.22 no.4
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    • pp.437-446
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    • 2012
  • A metagenomic library of cow rumen in the pCC1FOS phage vector was screened in $E.$ $coli$ EPI300 for cellulase activity on carboxymethyl cellulose agar plates. One clone was partially digested with $Sau$3AI, ligated into the $Bam$HI site of the pBluescript II SK+ vector, and transformed into $E.$ $coli$ $DH5{\alpha}$. We obtained a 1.5 kb insert DNA, designated $cel$5C, which hydrolyzes carboxymethyl cellulose. The cel5C gene has an open reading frame (ORF) of 1,125 bp encoding 374 amino acids. It belongs to the glycosyl hydrolase family 5 with the conserved domain LIMEGFNEIN. The molecular mass of the Cel5C protein induced from $E.$ $coli$ $DH5{\alpha}$, as analyzed by CMC SDS-PAGE, appeared to be approximately 42 kDa. The enzyme showed optimum cellulase activity at pH 4.0, and $50^{\circ}C$. We examined whether the $cel$5C gene comes from the 49 identified cow rumen bacteria using PCR. No PCR bands were identified, suggesting that the $cel$5C gene came from the unidentified cow rumen bacteria.