High level secretion of cellobiohydrolases by Saccharomyces cerevisiae

Table 2 Fusion genes created with T. emersonii cbh1 for expression in S. cerevisiae

Origin of CBM	Position attached	Expression plasmid	Recombinant yeast strain abbreviation	Primers used for construction (5'-3')
T. reesei cbh1 ^a	C-terminus	pMI529	Sc[Tecbh1-TrCBM-C]; M0759^c	399Trcbh1-L GCGACGAGTCAACCCTCCAGGTGGTAACAGAGGTACTACCAC 400Trcbh1-R GCGACTCGAGGGCGCGCCTACAAACATTGAGAGTAGTATGGGTTTA
H. grisea cbh1 ^a	C-terminus	pTeHg	Sc[Tecbh1-HgCBM-C]	Te-CBH-F TATAGAATTCTTAATTAAATGCTAAGAAGAGCTTTACTATTG Te-CBH-R TATACGTCTCTGGACCGAATTTAATGTTGGAGTA Hg-CBM-F TATACGTCTCGGTCCAATCGGTTCCACAGT Hg-CBM-R TATACTCGAGGCGCGCCTTATAAACATTGAGAGTACCAGTC
C. thermophilum cbh1 ^a	C-terminus	pMI566	Sc[Tecbh1-CtCBM-C]	392ENO1p-F CAGGATCCCAATTAATGTGAGTTACC 393TeCt-R ACAGTGGATCCGATTGGACCGAATTTAATGTTGG
T. reesei cbh2 ^a	N-terminus	pMI528	Sc[Tecbh1-TrCBM-N]	406TEM CBH1 NCBM-L CCTCCGAATTCATGCTAAGAAGAGCTTTACTATTGA-GCTCTTCTGCTATCTTGGCCGTTAAGGCTCAAGCCTGCTCCTCTGTTTGG 407TEM CBH1 NCBM-R AAACTTCAAGTCACGTGGACATTGAGAGTCACAG
T. reesei cbh2 ^b	N-terminus	pDLG117	Sc[Tecbh1-TrCBM-N2]	NCBM-L GAATTCATAATGGTCTCCTTC NCBM-R AAAGCTCTCGAGTTAAGAAGC NCBM-OL2 CGGTACCGGCTTGTTGAGAGTAAGTAGCAGTACCGG
T. reesei cbh1 ^b	C-terminus	pDLG118	Sc[Tecbh1-TrCBM-C2]	CCBM-L GAATTCATAATGGTCTCCTTC CCBM-R AATCAAAAGCTCTCGAGTTAC CCBM-OL2n GTTACCACCTGGAGGGTTAGAAGCAGTGAAAGTGGAG

^a Native secretion signal; ^bT. reesei xyn2 secretion signal; ^cdiploid ura3Δ/ura3Δ FUR1/fur1Δ strain which has a functional xylose pathway i.e. over expressed pentose pathway genes and Piromyces xylA, and gre3 deleted.

ISSN: 2731-3654