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Effect of site-directed mutagenesis of citB on the expression and activity of Bacillus subtilis aconitase.
Mikrobiologiia. 2010 Nov-Dec; 79(6):774-8.M

Abstract

Bacillus subtilis aconitase, encoded by the citB gene, is a bifunctional enzyme, which can not only interconvert citrate and isocitrate, but also has the RNA binding function similar to the eukaryotic protein IRP-1 (iron regulatory protein 1). Homology analysis between eukaryotic aconitase and B. subtilis aconitase indicates that the amino acids 741-745 probably have important function for the B. subtilis aconitase. To analyse the exact effect of these amino acids for aconitase activity, a site-directed mutagenesis of the citB is constructed, in which, the Arg741 and Gln745 are both changed into Glu. The resulting strain exhibits an increased enzymatic activity of aconitase comparing to that of the wild-type strain. Western blotting shows that the aconitase protein expression level is significantly increased in the mutant strain. By beta-galactosidase activity assay, the transcription level of citB is also increased. These results indicate that the mutation of citB gene has significant effect on B. subtilis aconitase transcription, expression and enzymatic activity.

Authors+Show Affiliations

Gao W
The Key Laboratory of Molecular Microbiology and Technology, Ministry of Education of China, 94 Weijin Road, College of Life Sciences, Nankai University, Tianjin 300071, China.
Dai S
No affiliation info available
Liu Q
No affiliation info available
Xu H
No affiliation info available
Bai Y
No affiliation info available
Qiao M
No affiliation info available

MeSH

Aconitate HydrataseAmino Acid SequenceAmino Acid SubstitutionBacillus subtilisBacterial ProteinsGene Expression Regulation, BacterialGenes, BacterialIsocitratesMolecular Sequence DataMutagenesis, Site-DirectedMutationSequence Homology, Nucleic AcidTranscription, Geneticbeta-Galactosidase

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

21446632

Citation

Gao, Weihua, et al. "Effect of Site-directed Mutagenesis of citB On the Expression and Activity of Bacillus Subtilis Aconitase." Mikrobiologiia, vol. 79, no. 6, 2010, pp. 774-8.
Gao W, Dai S, Liu Q, et al. Effect of site-directed mutagenesis of citB on the expression and activity of Bacillus subtilis aconitase. Mikrobiologiia. 2010;79(6):774-8.
Gao, W., Dai, S., Liu, Q., Xu, H., Bai, Y., & Qiao, M. (2010). Effect of site-directed mutagenesis of citB on the expression and activity of Bacillus subtilis aconitase. Mikrobiologiia, 79(6), 774-8.
Gao W, et al. Effect of Site-directed Mutagenesis of citB On the Expression and Activity of Bacillus Subtilis Aconitase. Mikrobiologiia. 2010 Nov-Dec;79(6):774-8. PubMed PMID: 21446632.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Effect of site-directed mutagenesis of citB on the expression and activity of Bacillus subtilis aconitase. AU - Gao,Weihua, AU - Dai,Sen, AU - Liu,Quanli, AU - Xu,Haijin, AU - Bai,Yanlin, AU - Qiao,Mingqiang, PY - 2011/3/31/entrez PY - 2011/3/31/pubmed PY - 2011/7/20/medline SP - 774 EP - 8 JF - Mikrobiologiia JO - Mikrobiologiia VL - 79 IS - 6 N2 - Bacillus subtilis aconitase, encoded by the citB gene, is a bifunctional enzyme, which can not only interconvert citrate and isocitrate, but also has the RNA binding function similar to the eukaryotic protein IRP-1 (iron regulatory protein 1). Homology analysis between eukaryotic aconitase and B. subtilis aconitase indicates that the amino acids 741-745 probably have important function for the B. subtilis aconitase. To analyse the exact effect of these amino acids for aconitase activity, a site-directed mutagenesis of the citB is constructed, in which, the Arg741 and Gln745 are both changed into Glu. The resulting strain exhibits an increased enzymatic activity of aconitase comparing to that of the wild-type strain. Western blotting shows that the aconitase protein expression level is significantly increased in the mutant strain. By beta-galactosidase activity assay, the transcription level of citB is also increased. These results indicate that the mutation of citB gene has significant effect on B. subtilis aconitase transcription, expression and enzymatic activity. SN - 0026-3656 UR - https://www.unboundmedicine.com/medline/citation/21446632/Effect_of_site_directed_mutagenesis_of_citB_on_the_expression_and_activity_of_Bacillus_subtilis_aconitase_ L2 - https://biocyc.org/gene?orgid=BSUB&id=BSU18000 DB - PRIME DP - Unbound Medicine ER -