Table 1 Plasmids and strains used in present study

Plasmids

pGAPZmazF

Vector for constitutive secreted protein expression; Zeo^r

Invitrogen

pPIC9K

Vector for inducible secreted protein expression; HIS4; kan^r & Amp^r

Invitrogen

pGAPZmazF (E,N)

pGAPZmazF based expression vector, containing EcoR I and Not I at multiple cloning site; Zeo^r

Our lab

pGAPZH

pGAPZ based expression vector, carrying the HIS4 fragment from pPIC9K; Zeo^r

Our lab

pGAPZ-XI-His

pGAPZ containing XI from Orpinomyces spp. and HIS4 fragment from pPIC9K; Zeo^r

This study

pGAPZ-XK

pGAPZ containing XK from P. pastoris GS115; Zeo^r

This study

pGAPKH-3Sman

pGAPKH containing three copies of alkaline β-mannanase gene, Kan^r

[33]

Strains

E. coli DH5α

Takara

P. pastoris GS115

his4⁻, Mut⁺

Invitrogen

Orpinomyces spp.

Our lab

GS-XI

GS115 integrated with the recombinant plasmid pGAPZ-XI-His

This study

GS-XK

GS115 integrated with the recombinant plasmid pGAPZ-XK

This study

GS-XI-XK

GS115 integrated with the recombinant plasmid pGAPZ-XI-His and pGAPZ-XK

This study

GS115^SB50

GS115 with 50 generations of evolution

This study

GS-XI^SB50

GS-XI with 50 generations of evolution

This study

GS-XK^SB50

GS-XK with 50 generations of evolution

This study

GS-XI-XK^SB50

GS-XI-XK with 50 generations of evolution

This study

GS-3Sman

GS115 integrated with the recombinant plasmid pGAPKH-3Sman

This study

GS-XI-3Sman

GS-XI^SB50 integrated with the recombinant plasmid pGAPKH-3Sman

This study