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Start Over Topic pichia pastoris Publication Year Range Last 50 years
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1. α-Fucosidases with different substrate specificities from two species of Fusarium

Author

Jonathan D. Walton, Janet M. Paper, John S. Scott-Craig, Mareike Bongers, Richard E. Wiemels, Ahmed Faik, David Cavalier, and Melissa S. Borrusch

Subjects
alpha-L-Fucosidase, Genetics, Subfamily, Sequence Homology, Amino Acid, Sequence analysis, Molecular Sequence Data, Fungal genetics, food and beverages, Sequence alignment, Sequence Analysis, DNA, General Medicine, Biology, biology.organism_classification, Applied Microbiology and Biotechnology, Substrate Specificity, Pichia pastoris, Microbiology, Fusarium, Fusarium oxysporum, Glycosides, DNA, Fungal, Sequence Alignment, Gene, Trichoderma reesei, Biotechnology
Abstract

Two fungal-secreted α-fucosidases and their genes were characterized. FoFCO1 was purified from culture filtrates of Fusarium oxysporum strain 0685 grown on l-fucose and its encoding gene identified in the sequenced genome of strain 4287. FoFCO1 was active on p-nitrophenyl-α-fucoside (pNP-Fuc), but did not defucosylate a nonasaccharide (XXFG) fragment of pea xyloglucan. A putative α-fucosidase gene (FgFCO1) from Fusarium graminearum was expressed in Pichia pastoris. FgFCO1 was ∼1,800 times less active on pNP-Fuc than FoFCO1, but was able to defucosylate the XXFG nonasaccharide. Although FgFCO1 and FoFCO1 both belong to Glycosyl Hydrolase family 29, they share

Published
2012

2. Secretory lipase from the human pathogen Leishmania major: Heterologous expression in the yeast Pichia pastoris and biochemical characterization

Author

Madiha Bou Ali, Youssef Gargouri, Saoussen Ben Ayed, Aymen Bali, Yassine Ben Ali, Dhafer Laouini, École Nationale d'Ingénieurs de Sfax | National School of Engineers of Sfax (ENIS), Université de Sfax - University of Sfax, Laboratoire de Transmission, Contrôle et Immunobiologie des Infections - Laboratory of Transmission, Control and Immunobiology of Infection (LR11IPT02), Institut Pasteur de Tunis, Réseau International des Instituts Pasteur (RIIP)-Réseau International des Instituts Pasteur (RIIP), and The authors would like to thank Mr. Anouar Fendri, Professor of English, who kindly proofread this paper.

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
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.

Published
2018