This study addresses the influence of upstream region sequence on the strength of has operon promoter in highly encapsulated S. equi subsp. zooepidemicus (SEZ). For this purpose, seven different strains were constructed. Each strain carries a point mutation in one of the following positions upstream of the has promoter: -43, -44, -49, and -50 bp. To facilitate measuring of the recombinant promoter relative strength, ß-glucuronidase gene was used as a reporter gene. Three mutations located in positions -49 and -50: AT, GT, and AG, positively impacted has promoter strength when compared to the wild type sequence GG. Conversely, two other mutations: TG and TT, exhibited a slight inhibitory effect. Further, three different strains carrying chromosomal mutations in the has promoter region were constructed. In two cases, the has operon is under the control of a stronger promoter and in the third strain the has operon is controlled by a weaker promoter. The laboratory fermenter scale cultivations confirmed the increase of hyaluronan yields for SEZPhasAG and SEZPhas2G, resulting 116 and 105 %, respectively. As expected, the yield of the hyaluronic acid of SEZPhas2B strain fell to 41 %.
A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding
Epidemiologic analysis of group A streptococcal serotypes associated with severe systemic infections, rheumatic fever, or uncomplicated pharyngitis
Specific binding of the activator Mga to promoter sequences of the emm and scpA genes in the group A streptococcus
Structural dissection and functional analysis of the complex promoter of the streptokinase gene from Streptococcus equisimilis H46A
Hyaluronic acid capsule and the role of streptococcal entry into keratinocytes in invasive skin infection
Relative contributions of hyaluronic acid capsule and M protein to virulence in a mucoid strain of the group A Streptococcus.
Role of putative virulence factors of Streptococcus pyogenes in mouse models of long-term throat colonization and pneumonia.
Structure of the has operon promoter and regulation of hyaluronic acid capsule expression in group A Streptococcus
Chlorella virus PBCV-1 encodes functional glutamine: fructose-6-phosphate amidotransferase and UDP-glucose dehydrogenase enzymes
The role of hyaluronic acid capsular material of Streptococcus equi subsp. zooepidemicus in mediating adherence to HeLa cells and in resisting phagocytosis
The streptococcal hyaluronan synthases are inhibited by sulfhydryl-modifying reagents, but conserved cysteine residues are not essential for enzyme function
Site-directed mutation of conserved cysteine residues does not inactivate the Streptococcus pyogenes hyaluronan synthase
Increase in hyaluronic acid production by Streptococcus equi subsp. zooepidemicus strain deficient in beta-glucuronidase in laboratory conditions
Mutation of two intramembrane polar residues conserved within the hyaluronan synthase family alters hyaluronan product size
Evolution of the hyaluronic acid synthesis (has) operon in Streptococcus zooepidemicus and other pathogenic streptococci.
Hyaluronan molecular weight is controlled by UDP-N-acetylglucosamine concentration in Streptococcus zooepidemicus.
Understanding plasmid effect on hyaluronic acid molecular weight produced by Streptococcus equi subsp. zooepidemicus
Structure of the has operon promoter and the effect of mutations on the has promoter strength in Streptococcus equi subsp. zooepidemicus
The role of hyaluronic acid precursor concentrations in molecular weight control in Streptococcus zooepidemicus
Bioinformatics in Biomedicine
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