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Proteomic Analysis of Bacillus Thuringiensis at Different Growth Phases by Using an Automated Online Two-dimensional Liquid Chromatography-tandem Mass Spectrometry Strategy

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Date 2012 May 29
PMID 22636013
Citations 14
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Abstract

The proteome of a new Bacillus thuringiensis subsp. kurstaki strain, 4.0718, from the middle vegetative (T(1)), early sporulation (T(2)), and late sporulation (T(3)) phases was analyzed using an integrated liquid chromatography (LC)-based protein identification system. The system comprised two-dimensional (2D) LC coupled with nanoscale electrospray ionization (ESI) tandem mass spectrometry (MS/MS) on a high-resolution hybrid mass spectrometer with an automated data analysis system. After deletion of redundant proteins from the different batches and B. thuringiensis subspecies, 918, 703, and 778 proteins were identified in the respective three phases. Their molecular masses ranged from 4.6 Da to 477.4 Da, and their isoelectric points ranged from 4.01 to 11.84. Function clustering revealed that most of the proteins in the three phases were functional metabolic proteins, followed by proteins participating in cell processes. Small molecular and macromolecular metabolic proteins were further classified according to the Kyoto Encyclopedia of Genes and Genome and BioCyc metabolic pathway database. Three protoxins (Cry2Aa, Cry1Aa, and Cry1Ac) as well as a series of potential intracellular active factors were detected. Many significant proteins related to spore and crystal formation, including sporulation proteins, help proteins, chaperones, and so on, were identified. The expression patterns of two identified proteins, CotJc and glutamine synthetase, were validated by Western blot analysis, which further confirmed the MS results. This study is the first to use shotgun technology to research the proteome of B. thuringiensis. Valuable experimental data are provided regarding the methodology of analyzing the B. thuringiensis proteome (which can be used to produce insecticidal crystal proteins) and have been added to the related protein database.

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References
1.
Fu Z, Sun Y, Xia L, Ding X, Mo X, Li X . Assessment of protoxin composition of Bacillus thuringiensis strains by use of polyacrylamide gel block and mass spectrometry. Appl Microbiol Biotechnol. 2008; 79(5):875-80. DOI: 10.1007/s00253-008-1488-0. View

2.
Gould G . History of science--spores. J Appl Microbiol. 2006; 101(3):507-13. DOI: 10.1111/j.1365-2672.2006.02888.x. View

3.
Fedhila S, Nel P, Lereclus D . The InhA2 metalloprotease of Bacillus thuringiensis strain 407 is required for pathogenicity in insects infected via the oral route. J Bacteriol. 2002; 184(12):3296-304. PMC: 135110. DOI: 10.1128/JB.184.12.3296-3304.2002. View

4.
Zhang L, Huang E, Lin J, Gelbic I, Zhang Q, Guan Y . A novel mosquitocidal Bacillus thuringiensis strain LLP29 isolated from the phylloplane of Magnolia denudata. Microbiol Res. 2009; 165(2):133-41. DOI: 10.1016/j.micres.2009.03.002. View

5.
Lovgren A, Zhang M, Engstrom A, Dalhammar G, Landen R . Molecular characterization of immune inhibitor A, a secreted virulence protease from Bacillus thuringiensis. Mol Microbiol. 1990; 4(12):2137-46. DOI: 10.1111/j.1365-2958.1990.tb00575.x. View