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Secretory lipase from the human pathogen Leishmania major: Heterologous expression in the yeast Pichia pastoris and biochemical characterization
- Source :
- Biochimie, Biochimie, Elsevier, 2018, 146, pp.119--126. ⟨10.1016/j.biochi.2017.12.002⟩
- Publication Year :
- 2018
- Publisher :
- HAL CCSD, 2018.
-
Abstract
- International audience; Leishmaniasis is a parasitic reticuloendotheliosis whose pathogen is a zooflagellate belonging to the genus Leishmania transmitted by the bite of an infected phlebotome. Recently, a unique secretory lipase from the human pathogen Leishmania donovani Ldlip3 has been identified and characterized. This lipase has a high identity with a putative triacylglycerol lipase of Leishmania major (Lmlip2). In the present study, Lmlip2 was expressed in the eukaryotic heterologous expression system Pichia pastoris as tagged enzyme of 308 amino acids. Maximal protein production was reached after 2 days of fermentation. Optimal Lmlip2 lipase activity was measured using the pH stat technique at pH 8 at 26 °C using vinyl esters and triacylglycerols (true lipids) as substrates. Moreover, biochemical characterization of Lmlip2 contained in culture supernatant, illustrates that L. major secreted lipase is active and stable at low temperatures especially 26°and prefer neutral pH; concerning substrate specificityLmlip2 presents a preference for short chains lipid substrates vinyl esters such as VC2, VC3 and VC4 likewise, it is capable to hydrolyze long chain triacylglycerols like olive oil. Metal ions and surfactants tested in this study decrease Lmlip2 activity. Further studies are needed to clarify the relation between the lipase activity and the virulence. Thus, it could lead to the identification of novel targets to block cutaneous Leishmaniasis in human hosts.
- Subjects :
- 0301 basic medicine
Models, Molecular
MESH: Leishmania major/genetics
MESH: Enzyme Stability
MESH: Hydrogen-Ion Concentration
Protein Conformation
Gene Expression
MESH: Amino Acid Sequence
MESH: Base Sequence
01 natural sciences
Biochemistry
Pichia
MESH: Protein Conformation
Enzyme Stability
MESH: Pichia/genetics
Leishmania major
MESH: Lipase/antagonists & inhibitors
Cloning, Molecular
Enzyme Inhibitors
Leishmania
MESH: Lipase/metabolism
biology
Chemistry
Temperature
General Medicine
Hydrogen-Ion Concentration
MESH: Temperature
Parasite
MESH: Biocatalysis
MESH: Models, Molecular
MESH: Gene Expression
Leishmania donovani
Triacylglycerol lipase
MESH: Sequence Alignment
MESH: Leishmania major/enzymology
Pichia pastoris
03 medical and health sciences
Humans
MESH: Cloning, Molecular
[SDV.MP.PAR]Life Sciences [q-bio]/Microbiology and Parasitology/Parasitology
[SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology
Amino Acid Sequence
Lipase
MESH: Humans
Base Sequence
MESH: Enzyme Inhibitors/pharmacology
010401 analytical chemistry
MESH: Lipase/chemistry
biology.organism_classification
Yeast
0104 chemical sciences
030104 developmental biology
MESH: Lipase/genetics
biology.protein
Biocatalysis
Heterologous expression
Sequence Alignment
Subjects
Details
- Language :
- English
- ISSN :
- 03009084
- Database :
- OpenAIRE
- Journal :
- Biochimie, Biochimie, Elsevier, 2018, 146, pp.119--126. ⟨10.1016/j.biochi.2017.12.002⟩
- Accession number :
- edsair.doi.dedup.....e746040dc0853d4dd4b53b5f22d4d85e