Engineering towards a complete heterologous cellulase secretome in Yarrowia lipolytica reveals its potential for consolidated bioprocessing

Table 2 Strains and plasmids used for expressing singular cellulolytic enzymes in this study

Expression plasmids	Plasmid description (species and GenBank accession number for expressed enzyme)	MW kDa	Source
pINA1296 (pYLSC1)	Hybrid promoter (hp4d); secretion signal (XPR2 pre- region); selection marker gene (LEU2)		Madzak et al., 2000
pNREL101	Tr EGII (P07982) in SfiI/XbaI cut pYLSC1	42	This study
pNREL102	Tr CBHII (P07987) in SfiI/HindIII cut pYLSC1	47	This study
CBHI constructs
pNREL106	Tr CBH1 (P62694) in SfiI/XbaI cut pYLSC1	52	This study
pNREL151	Chimeric CBHI (Te CBH1-Tr Linker-Tr CBM1; AAL89553 and P62694) in SfiI/Xbal cut pYLSC1	53	This study
pNREL152	Ct CBHI (CAM98448) in SfiI/Xbal cut pYLSC1	55	This study
pNREL153	Hg CBHI (CAA35159) in SfiI/Xbal cut pYLSC1	54	This study

The strain used for the enzyme expression was Y. lipolytica Po1g (MATa,leu2-270,ura3-302::URA3,xpr2-332,axp-2) (Madzak et al., 2000) [27]. The theoretic molecular weight (MW) was calculated based on amino acid sequences (without signal peptide). Abbreviation: CBH, cellobiohydrolase; Ct, Chaetomium thermophilum; EG, endoglucanase; Hg, Humicola grisea; Te, Talaromyces emersonii; Tr, T. reesei.

ISSN: 2731-3654