Cloning and Expression of Synthetic Genes Encoding the Broad Antimicrobial Spectrum Bacteriocins SRCAM 602, OR-7, E-760, and L-1077, by Recombinant Pichia pastoris

Table 1

Bacterial strains and plasmids used in this study.

Strain or plasmid	Description^a	Source and/or; reference^b
Strains
Enterococcus faecium T136	Enterocin A and B producer; MPA positive control	DNBTA
Pediococcus damnosus CECT4797	MPA indicator microorganism	CECT
Escherichia coli JM109	Selection of recombinant plasmids	Promega
Pichia pastoris X-33	Yeast producer	Invitrogen Life Technologies
Plasmids
pMA-T	Amp^r; carrier of synthetic genes	GeneArt Life Technologies
pPICZαA	Zeo^r; integrative plasmid carrying the secretion signal sequence from the S. cerevisiaeα-factor prepropeptide and functional sites for integration at the 5′ AOX1 locus of P. pastoris X-33	Invitrogen Life Technologies
pMATSRCAM602	Amp^r; pMA-T plasmid carrying the srcam602 synthetic gene with the P. pastoris codon usage	GeneArt Life Technologies
pMATOR-7	Amp^r; pMA-T plasmid carrying the or-7 synthetic gene with the P. pastoris codon usage	GeneArt Life Technologies
pMATE-760	Amp^r; pMA-T plasmid carrying the e-760 synthetic gene with the P. pastoris codon usage	GeneArt Life Technologies
pMATL-1077	Amp^r; pMA-T plasmid carrying the l-1077 synthetic gene with the P. pastoris codon usage	GeneArt Life Technologies
pSRCAM602	pPICZαA derivative with the srcam602 synthetic gene	This work
pOR-7	pPICZαA derivative with the or-7 synthetic gene	This work
pE-760	pPICZαA derivative with the e-760 synthetic gene	This work
pL-1077	pPICZαA derivative with the l-1077 synthetic gene	This work

Amp^r: ampicillin resistance; Zeo^r: zeocin resistance.
^bDNBTA: Departamento de Nutrición, Bromatología y Tecnología de los Alimentos, Facultad de Veterinaria, Universidad Complutense de Madrid (Madrid, Spain); CECT: Colección Española de Cultivos Tipo Valencia, Spain).