ABSTRACT
The lactic acid bacterium Tetragenococcus halophilus, which is used as a starter to brew soy sauce, comprises both cluster-forming strains and dispersed strains. The cluster-forming strains are industrially useful for obtaining clear soy sauce, because the cell clusters are trapped by filter cloth when the soy sauce mash is pressed. However, the molecular mechanism underlying cell cluster formation is unknown. Whole genome sequence analysis and subsequent target sequence analysis revealed that the cluster-forming strains commonly have functional defects in N-acetylglucosaminidase CseA, a peptidoglycan hydrolase. CseA is a multimodular protein that harbors a GH73 domain and six peptidoglycan-binding LysM domains. Recombinant CseA hydrolyzed peptidoglycan and promoted cell separation. Functional analysis of truncated CseA derivatives revealed that the LysM domains play an important role in efficient peptidoglycan degradation and cell separation. Taken together, the results of this study identify CseA as a factor that greatly affects the cluster formation in T. halophilus.
Subject(s)
Acetylglucosaminidase/metabolism , Bacterial Proteins/metabolism , Enterococcaceae/enzymology , Fermentation/genetics , Peptidoglycan/metabolism , Soy Foods/microbiology , Acetylglucosaminidase/chemistry , Acetylglucosaminidase/genetics , Bacterial Adhesion/genetics , Bacterial Proteins/chemistry , Bacterial Proteins/genetics , Cloning, Molecular , Enterococcaceae/classification , Enterococcaceae/genetics , Escherichia coli/genetics , Escherichia coli/metabolism , Food Analysis , Gene Expression , Genetic Complementation Test , Genetic Vectors/chemistry , Genetic Vectors/metabolism , Humans , Hydrolysis , Lactic Acid/biosynthesis , Peptidoglycan/chemistry , Protein Domains , Recombinant Proteins/chemistry , Recombinant Proteins/genetics , Recombinant Proteins/metabolism , Whole Genome SequencingABSTRACT
Individual olfactory sensory neurons express a single odorant receptor gene from either class I genes residing in a single cluster on a single chromosome or class II genes spread over multiple clusters on multiple chromosomes. Here, we identify an enhancer element for mouse class I genes, the J element, that is conserved through mammalian species from the platypus to humans. The J element regulates most class I genes expression by exerting an effect over ~ 3 megabases within the whole cluster. Deletion of the trans J element increases the expression frequencies of class I genes from the intact J allele, indicating that the allelic exclusion of class I genes depends on the activity of the J element. Our data reveal a long-range cis-regulatory element that governs the singular class I gene expression and has been phylogenetically preserved to retain a single cluster organization of class I genes in mammals."Each olfactory sensory neuron expresses a single odorant receptor gene from either class I or class II genes. Here, the authors identify an enhancer for mouse class I genes, that is highly conserved, and regulates most class I genes expression by acting over ~ 3 megabases within the whole cluster."
