Browse > Article

Two- Dimensional Electrophoresis Analysis of Proteins; Bacillus subtilis LTD and Its Antifungal Activity Deficient Mutant  

Lee, Young-Keun (Radiation Application Research Division, Korea Atomic Energy Research Institute)
Dinh, Le Thi (Institute for Nuclear Science and Techniques)
Jang, Yu-Sin (Radiation Application Research Division, Korea Atomic Energy Research Institute)
Chung, Hye-Young (Radiation Application Research Division, Korea Atomic Energy Research Institute)
Chang, Hwa-Hyoung (Radiation Application Research Division, Korea Atomic Energy Research Institute)
Publication Information
Korean Journal of Environmental Biology / v.22, no.4, 2004 , pp. 487-493 More about this Journal
Abstract
To investigate the antifungal activity related protein in pesticidal bacteria, a bacterial strain LTD was isolated from soil collected at Gimje in Jeonbuk province, Korea, and identified as Bacillus subtilis LTD based on a API50 CHB kit and 168 rDNA sequencing. It has an antifungal activity against 9 plant pathogenic fungi in a paper disc assay. The antifungal activity- deficient mutant, B. subtilis mLTD was induced at a 5 kGy dose of $^{60}Co$ gamma radiation. Using the two-dimensional electrophoresis and the matrix assisted laser desorption ionization time-of-flight mass spectrometry, the comparison analysis of proteins between the wild and mutant were performed. A major intracellular serine proteinase IspA (MW: 32.5 kDa), a NAD (P) H dehydrogenase (MW: 20.0 kDa), and a stage II sporulation protein AA, SpoIIAA (MW: 14.3kDa) were detected only in the B. subtilis LTD. These results suggested that the functions of these proteins found only in the B. subtilis LTD could. be closely related to the antifungal activity against plant pathogenic fungi.
Keywords
antifungal activity; bacillus subtilis LTD; gamma radiation; mutant; 2-DE;
Citations & Related Records
Times Cited By KSCI : 1  (Citation Analysis)
연도 인용수 순위
1 Cohen AM, K Rumpel, GH Coombs and JM Wastling. 2002. Characterisation of global protein expression by twodimensional electrophoresis and mass spectrometry: proteomics of Toxoplasma gondii. Int. J. Parasitol. 32: 39-51   DOI   ScienceOn
2 Emmert EAB and J Handelsman. 1999. Biocontrol of plant disease: a (Gram-) positive perspective. FEMS MicrobioI. Lett. 171:1-9   DOI   ScienceOn
3 Glick BR. 1995. The enhancement of plant growth by free living bacteria. Can. J. Microbiol. 41:109-117   DOI   ScienceOn
4 Herish S, K Manjula and AR Podile. 1998. Furarium udum is resistant to the mycolytic activity of a biocontrol strain of Bacillus subtilis AF 1. FEMS Microbiol. Ecol. 25:385-390   DOI   ScienceOn
5 Kim ST, KS Cho, YS Jang and KY Kang. 2001. Twodimensional electrophoretic analysis of rice proteins by polyethylene glycol fractionation for protein arrays. Electrophoresis 22:2103-2109   DOI   ScienceOn
6 Laemmli UK. 1970. Cleavage of structural proteins during assembly of the head of bacteriophage T4. Nature 277:680-685
7 Nordhoff E, V Egelhofer, P Giavalosco, H Eickhoff, M Horn, T Przewieslik, D Theiss, U Schneider, H Lehrach and J Gobom. 2001. Large-gel two-dimensional electrophoresis-matrix assisted laser desorption lionization-time of flight-mass spectrometry: an analytical challenge for studying complex protein mixtures. Electrophoresis 22:2844-2855   DOI   ScienceOn
8 O'Farrell PH. 1975. High resolution two-dimensional electrophoresis of proteins. J. BioI. Chern. 250:4007-402l
9 Phae CG and M Shoda. 1991. Investigation of optimal condition for foam separation of iturin, an antifungal peptide produced by Bacillus subtilis. J. Ferment. Bioeng. 71:118-121   DOI   ScienceOn
10 Thacker J. 1999. Repair of ionizing radiation damage in mammalian cells. Alternative pathways and their fidelity. C. R. Acad. Sci. 322:103-108   DOI   ScienceOn
11 Cho SJ, SK Lee, BJ Cha, YH Kim and KS Shin. 2003. Detection and characterization of the Gloeosporium gloeosporioides growth inhibitory compound iturin A from Bacillus subtilis strain KS03. FEMS Microbiol. Lett. 223:47- 51   DOI   ScienceOn
12 Hutchinson F. 1985. Chemical changes induced in DNA by ionizing radiation. Prog. Nucleic Acid Re. 32:115-154.   DOI
13 Lee Y-K, J-S Kim, H-Y Chung, Y-S Jang and B-1 Jang. 2003a. Two-dimensional electrophoresis analysis of proteins between Bacillus licheniformis DM3 and its antifungal deficient mutant. Kor. J. Environ. Agric. 22:203-209   DOI
14 Baker KF. 1987. Evolving concepts of biological control of plant pathogens. Ann. Rev. Phytopathol. 25:67 -85   DOI   ScienceOn
15 Lee Y-K, J-S Kim, Y-S Jang, K-S Cho and H-H Chang. 2003b. DNA microarray analysis of gene expression in antifungal bacterium of Bacillus lentimorbus WJ5. Kor. J. Microbiol. 39:141-147
16 Perkins DN, DJ Pappin, DM Ceasy and JS Cotrell. 1999. Probability-based protein identification by searching sequence databases using mass spectrometry data. Electrophoresis 20:3551-3567   DOI   ScienceOn
17 Strauch MA. 1993. Regulation of Bacillus subtilis gene expression during the transition from exponential growth to stationary phase. Prog. Nucleic Acid Re. 46:121-153   DOI
18 Fravel DR. 1988. Role of antibiosis in the biocontrol of diseases. Ann. Rev. Phytopathol. 26:75-91   DOI   ScienceOn
19 Tang WH. 1994. Yield-increasing bacteria (YIB) and biocontrol of sheath blight of rice. pp. 267-278. In MH Ryder, PM Stephens, and GD Bowen (eds.), Improving plant productivity with rhizosphere bacteria. Common Wealth Scientific and Industrial Research Organization, Adelaide
20 Leong J. 1986. Siderophores: their biochemistry and possible role in the biocontrol of plant pathogen. Ann. Rev. Phytopathol. 24:187-209   DOI   ScienceOn
21 Kerscher SJ. 2000. Diversity and origin of alternative NADH: ubiquinone oxidoreductases. Biochim. Biophys. Acta 1459:274-283   DOI   ScienceOn
22 Shoda M. 2000. Bacterial control of plant diseases. J. Biosci. Bioeng. 89:515-52l   DOI   PUBMED   ScienceOn
23 Chen FC, LF Shen, MC Tsai and KF Chak. 2003. The IspA protease's involvement in the regulation of the sporulation process of Bacillus thuringiensis is revealed by proteomic analysis. Biochem. Biophys. Res. Comm. 312:708-715   DOI   ScienceOn
24 Kloepper JW, R Lifshitz and RM Zablotowicz. 1989. Free living bacterial inocula for enhancing crop productivity. Trends Biotechnol. 7:9-43
25 Lee YK, HH Chang, JS Kim, JK Kim and KS Lee. 2000. Lignocellulolytic mutants of Pleurotus ostreatus induced by gamma-ray radJation and their genetic similarities. Rad. Phys. Chern. 57:145-150   DOI   ScienceOn
26 Phillips ZEV and MA Strauch. 2002. Bacillus subtilis sporulation and stationary phase gene expression. Cell. Mol. Life Sci. 59:392-402   DOI   ScienceOn
27 Woods RJ and AK Pikaev. 1994. Radiation dosimetry. pp.106-120. In Applied Radiation Chemistry: Radiation Processing. John Wiley & Sons, Inc., New York
28 Baum JA and T Malvar. 1995. Regulation of insecticidal crystal protein production in Bacillus thuringiensis. Mol. Microbiol. 18:1-12   DOI   ScienceOn