• Title/Summary/Keyword: ChiCW

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Analysis of the Involvement of Chitin-Binding Domain of ChiCW in Antifungal Activity, and Engineering a Novel Chimeric Chitinase with High Enzyme and Antifungal Activities

  • Huang, Chien-Jui;Guo, Shu-Huei;Chung, Shu-Chun;Lin, Yu-Ju;Chen, Chao-Ying
    • Journal of Microbiology and Biotechnology
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    • v.19 no.10
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    • pp.1169-1175
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    • 2009
  • An antifungal chitinase, ChiCW, produced by Bacillus cereus 28-9 is effective against conidial germination of Botrytis elliptica, the causal agent of lily leaf blight. ChiCW as a modular enzyme consists of a signal peptide, a catalytic domain, a fibronectin type-III-like domain, and a chitin-binding domain. When two C-terminal domains of ChiCW were truncated, $ChiCW{\Delta}FC$ (lacking the chitin-binding domain and fibronectin type III-like domain) lost its antifungal activity. Since $ChiCW{\Delta}C$ (lacking the chitin-binding domain) could not be expressed in Escherichia coli as $ChiCW{\Delta}FC$ did, a different strategy based on protein engineering technology was designed to investigate the involvement of the chitin-binding domain of ChiCW ($ChBD_{ChiCW}$) in antifungal activity in this study. Because ChiA1 of Bacillus circulans WL-12 is a modular enzyme with a higher hydrolytic activity than ChiCW but not inhibitory to conidial germination of Bo. elliptica and the similar domain composition of ChiA1 and ChiCW, the C-terminal truncated derivatives of ChiA1 were generated and used to construct chimeric chitinases with $ChBD_{ChiCW}$. When the chitin-binding domain of ChiA1 was replaced with $ChBD_{ChiCW}$, the chimeric chitinase named ChiAAAW exhibited both high enzyme activity and antifungal activity. The results indicate that $ChBD_{ChiCW}$ may play an important role in the antifungal activity of ChiCW.

Functions of the C-Terminal Region of Chitinase ChiCW from Bacillus cereus 28-9 in Substrate-Binding and Hydrolysis of Chitin

  • Huang, Chien-Jui;Chen, Chao-Ying
    • Journal of Microbiology and Biotechnology
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    • v.16 no.12
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    • pp.1897-1903
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    • 2006
  • In order to investigate the functions of the C-terminal region of chitinase ChiCW of Bacillus cereus 28-9, a C-terminal truncated enzyme, ChiCW$\Delta$FC, was expressed in Escherichia coli and purified to homogeneity for biochemical characterization. Compared with ChiCW, ChiCW$\Delta$FC exhibited higher chitinase activity at high temperature and pH, but expressed lower hydrolytic and binding activities toward insoluble substrates. In addition, kinetic properties indicated that ChiCW$\Delta$MC hydrolyzed oligomeric and polymeric substrates less efficiently than ChiCW. These results suggest that the C-terminal region of ChiCW plays important roles in substrate binding and hydrolysis of chitin. In addition, the biological meaning of C-terminal proteolytic modification of ChiCW is discussed.

Synergistic Interactions Between Chitinase ChiCW and Fungicides Against Plant Fungal Pathogens

  • Huang, Chien-Jui;Chen, Chao-Ying
    • Journal of Microbiology and Biotechnology
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    • v.18 no.4
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    • pp.784-787
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    • 2008
  • Antifungal activity of ChiCW and synergistic interactions between ChiCW with fungicides were investigated. Conidial germinations of phytopathogenic fungi, Alternaria brassicicola, Botrytis elliptica, and Colletotrichum gloeosporioides, were inhibited by ChiCW but A. longipes was not. In addition, ChiCW showed synergistic effect with fungicides Switch (cyprodinil+fludioxonil) and tebuconazole to inhibit fungal conidial germinations. The level of synergism of ChiCW with tebuconazole was higher than that with Switch. The results indicate that ChiCW may exhibit a higher level of synergism with fungicides that have a primary effect upon membranes.

Identification of an Antifungal Chitinase from a Potential Biocontrol Agent, Bacillus cereus 28-9

  • Huang, Chien-Jui;Wang, Tang-Kai;Chung, Shu-Chun;Chen, Chao-Ying
    • BMB Reports
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    • v.38 no.1
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    • pp.82-88
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    • 2005
  • Bacillus cereus 28-9 is a chitinolytic bacterium isolated from lily plant in Taiwan. This bacterium exhibited biocontrol potential on Botrytis leaf blight of lily as demonstrated by a detached leaf assay and dual culture assay. At least two chitinases (ChiCW and ChiCH) were excreted by B. cereus 28-9. The ChiCW-encoding gene was cloned and moderately expressed in Escherichia coli DH5$\alpha$. Near homogenous ChiCW was obtained from the periplasmic fraction of E. coli cells harboring chiCW by a purification procedure. An in vitro assay showed that the purified ChiCW had inhibitory activity on conidial germination of Botrytis elliptica, a major fungal pathogen of lily leaf blight.

Clinical features of the gingiva according to maxillary anterior teeth form in adult (성인 상악 전치 형태에 따른 치은의 임상적 소견)

  • An, Chi-Hyun;Heo, Soo-Rye;Cho, Ik-Hyun;Kim, Hyung-Seop
    • Journal of Periodontal and Implant Science
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    • v.35 no.2
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    • pp.359-369
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    • 2005
  • It has been suggested that morphologic characteristics of the periodontium are partly related to the shape and form of the teeth. Furthermore, the severity of symptoms of periodontal disease have been proposed to differ among these various morphologic entities or "biotypes". The purpose of the present study was to examine the relationship between the form of the crowns in the maxillary anterior tooth segment and (1) a group of morphological characteristics and (2) the thickness of the gingiva. The thickness of gingiva was measured by ultrasonic device(SDM). 100 subjects devoid of symptoms of destructive periodontal disease were examined regarding, e.g., probing depth, gingival recession, width of keratinized gingiva, thickness of the keratinized gingiva. From maxillary study cast, the width(at the apical third-CW) and the length(CL) of the crowns of the 6 anterior teeth were determined. A CW/CL-ratio was calculated for each tooth and averaged for each tooth region. The individual mean CW/CL-ratio values for the central incisors were ranked. The 10 subjects ranked highest and the 10 ranked lowest were selected as having either a long-narrow(group N) or a short-wide(group W) form of the crown of the tooth. The data for each of the examined parameters were averaged for each tooth region in each subject and mean values for subjects in groups W and N were compared using the Student t-test. Stepwise multiple regression analysis, including data from the whole sample, was performed for each tooth region with the thickness of the free gingiva as the dependent variable. The results from the analyses demonstrated that individuals with a long-narrow form of the central incisors displayed, compared to individuals with a short-wide crown, form (l) a narrow zone of keratinized gingiva, (2) a pronounced "scalloped" contour of the gingival margin. There was no significant difference between groups N and W with respect to the thickness of the keratinized gingiva. The CW/CL-ratio data revealed that a certain form of the crowns in the central incisors was accompanied by a similar form in the lateral incisors and canine tooth region. The regression analyses demonstrated that the thickness of the keratinized gingiva in central, lateral incisors and canines was significantly related to the width of the keratinized gingiva.

An Estimation on the Field Application of Consolidants According to Rock Quality (암석 재질에 따른 보존처리제의 현장 적용성 평가)

  • Kim, Jae-Hwan;Han, Min-Su;Lee, Jang-Jon;Song, Chi-Young;Lee, Jae-Man;Kim, Min-Ji;Lee, Myeong-Sung
    • 보존과학연구
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    • s.30
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    • pp.79-91
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    • 2009
  • Stone cultural heritages are mostly situated in field, so they are damaged from mechanical, chemical and biological weathering, and their degree of strength is also weaken. Therefore, the scientific conservation of the stone cultural heritages are necessary in order to the long-term maintenance and safety conservation. In this study, we attempt to estimate on the field application of consolidants which are Wacker OH 100, Remmers KSE 300 and 1T1G according to the quality of the rocks. Based on the ultrasonic velocity, conditions of weathering damage before treatment are mainly ranked completely weathering condition (CW) or partly highly weathering condition (HW). After the first treatment, weathering damage conditions are changed by the high weathering condition (HW). These results shows that the internal pores of stone are filled with consolidant materials, so average ultrasonic velocities increases after treatment. And Remmers KSE 300 which is consolidant material is highest rate of increase of average ultrasonic velocities.

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