Single-Step Partial Purification of Intracellular β-Galactosidase from Kluyveromyces lactis Using Microemulsion Droplets.

Partial purification of β-galactosidase from the crude extract of Kluyveromyces lactis was carried out using water-in-isooctane microemulsions formed by the anionic surfactant, sodium di-ethylhexyl sulfosuccinate (Aerosol OT). In order to obtain the crude extract, yeast cells of K. lactis were disrupted by a cell disrupter and separated.

Characterization of Yarrowia lipolytica XPR2 multi-copy strains over-producing alkaline extracellular protease - a system for rapidly increasing secretory pathway cargo loads.

Upon transfer to alkaline extracellular protease (AEP) induction medium, strain 773-2 (50 integrated copies of XPR2), derived from highly inbred strain E129, grew for at least 10 h before AEP production began, and then growth rate decreased before increasing again; by then, cells had lost copies of XPR2 (Le Dall et al., 1994). Slowing of growth following AEP induction suggested that increased secretory pathway cargo load was affecting cell growth and that such a system had potential for secretion stress studies.

Isolation and characterization of the TRP1 gene from the yeast Yarrowia lipolytica and multiple gene disruption using a TRP blaster.

The TRP1 gene encoding N-(5'-phosphoribosyl)-anthranilate isomerase was isolated from the yeast Yarrowia lipolytica, in which only a few genetic marker genes are available. The Y. lipolytica TRP1 gene (YlTRP1) cloned by complementation of Y.

Dipeptidyl aminopeptidase processing and biosynthesis of alkaline extracellular protease from Yarrowia lipolytica.

Alkaline extracellular protease (AEP) from Yarrowia lipolytica is synthesized as a precursor with a 157 aa prepro-region. Signal peptide cleavage was shown to occur after Ala15 by N-terminal amino acid radiosequencing of the largest intracellular AEP precursor. AEP proteolytic activity was not required for AEP processing.