(Below N is a link to NCBI taxonomic web page and E link to ESTHER at designed phylum.) > cellular organisms: NE > Bacteria: NE > Proteobacteria: NE > Gammaproteobacteria: NE > Moraxellales: NE > Moraxellaceae: NE > Psychrobacter: NE > Psychrobacter celer: NE
LegendThis sequence has been compared to family alignement (MSA) red => minority aminoacid blue => majority aminoacid color intensity => conservation rate title => sequence position(MSA position)aminoacid rate Catalytic site Catalytic site in the MSA MTLSLYIRAALTPRTFLPILGSISTGVLLMTMQTISAQAAGSFYNCANAT GCQKVDSKWWTSSHTDTKYPLVMTHGLVGFTKLLGIDYFYGIPQTLMKGG SEVYATKTSSFNDSRVRGEQLLRQVKTISAVSGSPKVNLLGHSHGGHDIR YVAGVAPQYVASVTAVASPGQGAALADWVVEQIDNDRINGEYNTATKAAL TLFGFLGDFMDLNAGIPIDELQDQDGFGAVTALTTDYTKDFNREFSAAMP TSYCGQPTAIKVNGISYYSFSGVDQVTNVLDPTDYMMLATSQAYDDPNDG LVSACSSRLGYVIRDDYKMNHLDSVNQLLGLVAWGQPNPKTVYRTQVNRL KNANL
The lipase gene from Psychrobacter celer PU3 was cloned into pET-28a(+) expression vector and overexpressed in E. coli BL21 (DE3) pLysS cells. The purified Psychrobacter celer lipase (PCL) was characterized as an alkaline active enzyme and has a molecular mass of around 30 kDa. The PCL was active even at a low temperature and the optimum range was observed between 10 and 40 degreesC temperatures. MALDI-TOF and phylogenetic analysis ensued that Psychrobacter celer PU3 lipase (PCL) was closely related to P. aureginosa lipase (PAL). MD simulation results suggests that temperature change did not affect overall structure of PCL, but it may alter temperature- dependent PCL structural changes. R(1) (129-135 AA) and R(2) (187-191 AA) regions could be important for temperature-dependent PCL function as they fluctuate much at 35 degreesC temperature. PMSF completely inhibited PCL lipase activity and it demonstrates the presence of serine residues in the active site of PCL. PCL is moderately halophilic and most of the tested organic solvents found to be inhibiting the lipase activity except the solvents ethanol and methanol. PCL activity was increased with surfactants (SDS and CTAB) and bleaching agents (hydrogen peroxide). The effect of different metal ions on PCL resulted that only mercuric chloride was found as the enhancer of the lipase activity. Antibiofilm property of PCL was evaluated against pathogenic Vibrio parahaemolyticus isolated from the diseased shrimp and MIC value was 500 U. PCL significantly altered the morphology and biofilm density of V. parahaemolyticus and the same was observed through scanning electron microscope (SEM) and confocal laser scanning microscope (CLSM) imaging. RT-PCR analysis revealed that the mRNA expression level of biofilm, colony morphology and major toxin-related (aphA, luxS, opaR, tolC, toxR) genes of V. parahaemolyticus were significantly downregulated with PCL treatment.
