title: Yeast Kluyveromyces lactis as host for expression of the bacterial lipase: cloning and adaptation of the new lipase gene from Serratia sp.
abstract: Many microbial lipases have been successfully expressed in yeasts, but not in industrially attractive Kluyveromyces lactis, which among other benefits can be cultivated on a medium supplemented with whey--cheap and easily available industrial waste. A new bacterial lipase from Serratia sp. was isolated and for the first time expressed into the yeast Kluyveromyces lactis by heterologous protein expression system based on a strong promoter of Kluyveromyces marxianus triosephosphate isomerase gene and signal peptide of Kluyveromyces marxianus endopolygalacturonase gene. In addition, the bacterial lipase gene was synthesized de novo by taking into account a codon usage bias optimal for K. lactis and was expressed into the yeast K. lactis also. Both resulting strains were characterized by high output level of the target protein secreted extracellularly. Secreted lipases were characterized for activity and stability.
authors.affiliation: Sector of Applied Biocatalysis, Institute of Biotechnology, Vilnius University, V.A. Graičiūno str. 8, 02241, Vilnius, Lithuania.
authors.forename: Rimantas
authors.initials: R
authors.lastname: Šiekštelė
authors.affiliation: Sector of Applied Biocatalysis, Institute of Biotechnology, Vilnius University, V.A. Graičiūno str. 8, 02241, Vilnius, Lithuania.
authors.forename: Aušra
authors.initials: A
authors.lastname: Veteikytė
authors.affiliation: JSC Biocentras, V.A. Graičiūno str. 10, 02241, Vilnius, Lithuania.
authors.forename: Bronius
authors.initials: B
authors.lastname: Tvaska
authors.affiliation: Sector of Applied Biocatalysis, Institute of Biotechnology, Vilnius University, V.A. Graičiūno str. 8, 02241, Vilnius, Lithuania. inga.matijosyte@bti.vu.lt.
authors.forename: Inga
authors.initials: I
authors.lastname: Matijošytė
mesh_terms: D003001:Cloning, Molecular; D003062:Codon / Q000235:genetics; D004795:Enzyme Stability; D005798:Genes, Bacterial / Q000235:genetics; D005818:Genetic Engineering; D007716:Kluyveromyces / Q000235:genetics* / Q000378:metabolism; D008049:Lipase / Q000235:genetics* / Q000378:metabolism*; D011096:Polygalacturonase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D011401:Promoter Regions, Genetic / Q000235:genetics; D021382:Protein Sorting Signals / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D012705:Serratia / Q000201:enzymology* / Q000235:genetics*; D014305:Triose-Phosphate Isomerase / Q000235:genetics
chemical_list: D003062:Codon; D021382:Protein Sorting Signals; D011993:Recombinant Fusion Proteins; C081378:solysime; D008049:Lipase; D011096:Polygalacturonase; D014305:Triose-Phosphate Isomerase
keywords: Kluyveromyces lactis; Lipase; Serratia sp.; Whey; de novo synthesis