Acquisition of flocculation phenotype by Kluyveromyces marxianus when overexpressing GAP1 gene encoding an isoform of glyceraldehyde-3-phosphate dehydrogenase.
The use of flocculating yeast strains has been considered as a convenient approach to obtain high cell densities in bioreactors with increasing productivity in continuous operations. In Kluyveromyces marxianus ATTC 10022, the GAP1 gene encodes an isoform of glyceraldehyde-3-phosphate dehydrogenase-p37-that is accumulated in the cell wall and is involved in flocculation. To test the use of p37 as a tool for engineering Kluyveromyces cells to display a flocculation phenotype, K. marxianus CCT 3172 was transformed with an expression vector containing GAP1. This vector is based on the pY37 previously described, harbouring a S11 Kluyveromyces origin of replication, and the expression of GAP1 is under the control of GAL1. Kluyveromyces cells overexpressing GAP1 acquired a flocculent phenotype together with the accumulation of p37 in the cell wall. The results support the use of GAP1 gene as a molecular tool for inducing flocculation.
Almeida
Catarina
C
Instituto de Biologia Molecular e Celular, Universidade do Porto, Rua do Campo Alegre, 823, 4150-180, Porto, Portugal.
Queirós
Odília
O
Wheals
Alan
A
Teixeira
José
J
Moradas-Ferreira
Pedro
P
D015152:Blotting, Northern; D015153:Blotting, Western; D002473:Cell Wall / Q000235:genetics* / Q000502:physiology; D005427:Flocculation; D005656:Fungal Proteins / Q000235:genetics* / Q000502:physiology; D005822:Genetic Vectors; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism; D007527:Isoenzymes / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism; D007716:Kluyveromyces / Q000201:enzymology* / Q000235:genetics / Q000502:physiology*; D016133:Polymerase Chain Reaction; D012331:RNA, Fungal / Q000737:chemistry / Q000235:genetics; D014170:Transformation, Genetic / Q000502:physiology*
D005656:Fungal Proteins; D007527:Isoenzymes; D012331:RNA, Fungal; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases