The phospholipase activity of Staphylococcus hyicus lipase strongly depends on a single Ser to Val mutation

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• 作者：van Kampen ; M.D. ; Simons ; J.-W.F.A. ; Dekker ; N. ; Egmond ; M.R. ; Verheij ; H.M.
• 关键词：Phospholipase activity ; Staphylococcus hyicus ; C-terminal ; GPLRP2 ; guinea pig lipase related protein 2 ; SHL ; Staphylococcus hyicus lipase ; SAL ; Staphylococcus aureus lipase ; SEL ; Staphylococcus epidermidis lipase ; PGL ; Pseudomonas glumae lipase ; C16thioPC
• 刊名：Chemistry and Physics of Lipids
• 出版年：1998
• 出版时间：June, 1998
• 年：1998
• 卷：93
• 期：1-2
• 页码：39-45
• 全文大小：91.5 K

文摘

Site-directed mutagenesis and domain exchange were used to investigate the role of the C-terminal domains of Staphylococcus hyicus lipase (SHL) and S. aureus lipase (SAL) in substrate selectivity. The introduction of a single point mutation coding for the substitution of Val for Ser356 in SHL yields an enzyme which has retained full lipase activity, although with more than 12-fold lower phospholipase activity. Starting with this S356V variant of SHL the C-terminal 40 amino acids were replaced by the corresponding SAL sequence. Although 23 amino acid changes were introduced simultaneously the impact on the phospholipase/lipase activity ratio was only 4-fold. We therefore conclude that in the C-terminal domain it is Ser356 which mainly determines phospholipase activity. The introduction of a Val357 to Ser substitution in SAL did not turn SAL into a phospholipase, showing that residues from other domains contribute to this activity as well. The results are discussed in view of the sequence homology of lipases and (lyso)phospholipases.