Biochemical and molecular characterization of Staphylococcus xylosus lipase.
The Staphylococcus xylosus strain secretes a non-induced lipase in culture medium: S. xylosus lipase (SXL). Pure SXL is a monomeric protein (43 kDa). The 23 N-terminal amino acid residues were sequenced. This sequence is identical to that of Staphylococcus simulans lipase (SSL); in addition, it exhibits a high degree of homology with Staphylococcus aureus lipase (SAL NCTC 8530) sequences. The cloning and sequencing of gene part encoding the mature lipase shows one nucleotide difference with SSL, which corresponds to the change of one residue at a position 311. The lipase activity is maximal at pH 8.2 and 45 degrees C. SXL is able to hydrolyse triacylglycerols without chain length specificity. The specific activity of about 1900 U/mg was measured using tributyrin or triolein as substrate at pH 8.2 and at 45 degrees C in the presence of 2 mM CaCl2. In contrast to some previously characterized staphylococcal lipases, Ca2+ is not required to trigger the activity of SXL. SXL was found to be stable between pH 5 and pH 8.5. The enzyme maintains 50% of its activity after a 15-min incubation at 60 degrees C. Using tripropionin or vinyl esters as substrates, SXL does not present the interfacial activation phenomenon. Unlike many lipases, SXL is able to hydrolyse its substrate in the presence of bile salts or amphiphilic proteins. SXL is a serine enzyme, which is inhibited by THL.
Mosbah
Habib
H
Laboratoire de biochimie et de génie enzymatique des lipases, ENIS, BPW 3038 Sfax, Tunisia.
Sayari
Adel
A
Mejdoub
Hafedh
H
Dhouib
Hafedh
H
Gargouri
Youssef
Y
D000595:Amino Acid Sequence; D001483:Base Sequence; D001647:Bile Acids and Salts / Q000494:pharmacology; D002118:Calcium / Q000494:pharmacology; D003001:Cloning, Molecular; D004655:Emulsions; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D007783:Lactones / Q000494:pharmacology; D008049:Lipase / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D008969:Molecular Sequence Data; D000077403:Orlistat; D013210:Staphylococcus / Q000201:enzymology*; D013696:Temperature
D001647:Bile Acids and Salts; D004655:Emulsions; D007783:Lactones; D000077403:Orlistat; D008049:Lipase; D002118:Calcium