Limits...
LytM Fusion with SH3b-Like Domain Expands Its Activity to Physiological Conditions

View Article: PubMed Central - PubMed

ABSTRACT

Staphylococcus aureus remains one of the most common and at the same time the most dangerous bacteria. The spreading antibiotic resistance calls for intensification of research on staphylococcal physiology and development of new strategies for combating this threatening pathogen. We have engineered new chimeric enzymes comprising the enzymatically active domain (EAD) of autolysin LytM from S. aureus and the cell wall binding domain (CBD) from bacteriocin lysostaphin. They display potent activity in extended environmental conditions. Our results exemplify the possibility of exploring autolytic enzymes in engineering lysins with desired features. Moreover, they suggest a possible mechanism of autolysin physiological activity regulation by local ionic environments in the cell wall.

No MeSH data available.


Related in: MedlinePlus

Enzyme activity kinetics in low temperature. Studied enzymes were incubated with S. aureus NCTC 8325-4 reference strain in 50 mM glycine buffer, pH 8.0, supplemented with 100 mM NaCl (except for LytM_EAD) on ice for 1 hr and their activity was observed as a turbidity reduction, OD595 was measured every 10 min.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC5036312&req=5

f4: Enzyme activity kinetics in low temperature. Studied enzymes were incubated with S. aureus NCTC 8325-4 reference strain in 50 mM glycine buffer, pH 8.0, supplemented with 100 mM NaCl (except for LytM_EAD) on ice for 1 hr and their activity was observed as a turbidity reduction, OD595 was measured every 10 min.

Mentions: We have previously shown that, in contrast to lysostaphin, enzymatically active domain of LytM is able to lyse bacteria over a wide range of temperatures from 0°C to 45°C. At 4°C, LytM_EAD is over four times more active than lysostaphin (European Patent No. 2699254). We have tested whether Chimera remains as active at 4°C as its parental LytM domain. Surprisingly, it turned out that the fusion of CBD domain with LytM_EAD strongly reduces the activity of chimeric enzyme at low temperature compared to LytM_EAD. Moreover, the chimeric enzyme was even less active than Lss (Fig. 4).


LytM Fusion with SH3b-Like Domain Expands Its Activity to Physiological Conditions
Enzyme activity kinetics in low temperature. Studied enzymes were incubated with S. aureus NCTC 8325-4 reference strain in 50 mM glycine buffer, pH 8.0, supplemented with 100 mM NaCl (except for LytM_EAD) on ice for 1 hr and their activity was observed as a turbidity reduction, OD595 was measured every 10 min.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC5036312&req=5

f4: Enzyme activity kinetics in low temperature. Studied enzymes were incubated with S. aureus NCTC 8325-4 reference strain in 50 mM glycine buffer, pH 8.0, supplemented with 100 mM NaCl (except for LytM_EAD) on ice for 1 hr and their activity was observed as a turbidity reduction, OD595 was measured every 10 min.
Mentions: We have previously shown that, in contrast to lysostaphin, enzymatically active domain of LytM is able to lyse bacteria over a wide range of temperatures from 0°C to 45°C. At 4°C, LytM_EAD is over four times more active than lysostaphin (European Patent No. 2699254). We have tested whether Chimera remains as active at 4°C as its parental LytM domain. Surprisingly, it turned out that the fusion of CBD domain with LytM_EAD strongly reduces the activity of chimeric enzyme at low temperature compared to LytM_EAD. Moreover, the chimeric enzyme was even less active than Lss (Fig. 4).

View Article: PubMed Central - PubMed

ABSTRACT

Staphylococcus aureus remains one of the most common and at the same time the most dangerous bacteria. The spreading antibiotic resistance calls for intensification of research on staphylococcal physiology and development of new strategies for combating this threatening pathogen. We have engineered new chimeric enzymes comprising the enzymatically active domain (EAD) of autolysin LytM from S. aureus and the cell wall binding domain (CBD) from bacteriocin lysostaphin. They display potent activity in extended environmental conditions. Our results exemplify the possibility of exploring autolytic enzymes in engineering lysins with desired features. Moreover, they suggest a possible mechanism of autolysin physiological activity regulation by local ionic environments in the cell wall.

No MeSH data available.


Related in: MedlinePlus