Identification of a Gene Cluster Enabling Lactobacillus casei BL23 To Utilize myo-Inositol

• Published in: Applied and Environmental Microbiology Vol. 73; no. 12; pp. 3850 - 3858
• Main Authors: Yebra, María Jesús, Zúñiga, Manuel, Beaufils, Sophie, Pérez-Martínez, Gaspar, Deutscher, Josef, Monedero, Vicente
• Format: Journal Article
• Language: English
• Published: Washington, DC American Society for Microbiology 01.06.2007
• Subjects: Bacteria; Bacteriology; Base Sequence; Biochemistry, Molecular Biology; Biological and medical sciences; Blotting, Northern; Carboxy-Lyases - genetics; Carboxy-Lyases - metabolism; Deoxyribonucleic acid; DNA; DNA Primers - genetics; Food Microbiology; Fundamental and applied biological sciences. Psychology; Gene Expression Regulation, Bacterial - genetics; Genes; Genes, Bacterial - genetics; Genomics; Inositol - metabolism; Lactobacillus casei; Lactobacillus casei - genetics; Life Sciences; Likelihood Functions; Microbiology; Models, Genetic; Molecular biology; Molecular Sequence Data; Multigene Family - genetics; Operon - genetics; Phylogeny; Repressor Proteins - genetics; Sequence Analysis, DNA; Gene cluster; Lactic acid bacteria; Inositol; Lactobacillus casei; Bacteria; Lactobacillaceae; Identification; CARBON CATABOLITE REPRESSION; MYO-INOSITOL; IOLTABCDG1G2EJK CLUSTER; CATABOLITE CONTROL PROTEIN A; TRANSCRIPTIONAL FACTOR
• ISSN: 0099-2240; 1098-5336
• DOI: 10.1128/AEM.00243-07

Genome analysis of Lactobacillus casei BL23 revealed that, compared to L. casei ATCC 334, it carries a 12.8-kb DNA insertion containing genes involved in the catabolism of the cyclic polyol myo-inositol (MI). Indeed, L. casei ATCC 334 does not ferment MI, whereas strain BL23 is able to utilize this carbon source. The inserted DNA consists of an iolR gene encoding a DeoR family transcriptional repressor and a divergently transcribed iolTABCDG1G2EJK operon, encoding a complete MI catabolic pathway, in which the iolK gene probably codes for a malonate semialdehyde decarboxylase. The presence of iolK suggests that L. casei has two alternative pathways for the metabolism of malonic semialdehyde: (i) the classical MI catabolic pathway in which IolA (malonate semialdehyde dehydrogenase) catalyzes the formation of acetyl-coenzyme A from malonic semialdehyde and (ii) the conversion of malonic semialdehyde to acetaldehyde catalyzed by the product of iolK. The function of the iol genes was verified by the disruption of iolA, iolT, and iolD, which provided MI-negative strains. By contrast, the disruption of iolK resulted in a strain with no obvious defect in MI utilization. Transcriptional analyses conducted with different mutant strains showed that the iolTABCDG1G2EJK cluster is regulated by substrate-specific induction mediated by the inactivation of the transcriptional repressor IolR and by carbon catabolite repression mediated by the catabolite control protein A (CcpA). This is the first example of an operon for MI utilization in lactic acid bacteria and illustrates the versatility of carbohydrate utilization in L. casei BL23.