After construction, all expression vectors were verified
by DNA sequencing (GATC Biotech, Konstanz,
Germany). For Y. lipolytica transformation, vectors were
digested using NotI, thus generating a linear DNA with
Zeta sequences at both extremities, and purified. Then
the linear DNA fragments were introduced into the Zeta
docking platform of Y. lipolytica JMY1212 Zeta or randomly
into the genome of ∆pox strain using the lithium
acetate method [49]. Transformants were tested for
β-glucosidase activity on YNB glucose plate containing
pNP-βGlc and for growth on cellobiose using solid YNB
cellobiose plates. Clones displaying both activities were
retained for further analysis.