TABLE 4.

Production and antimicrobial activity of recombinant EntL50A and EntL50B from an S. cerevisiae L50A-20(pYABD02) and S. cerevisiae L50B-4(pYABD03) coculture^a

Time (h)	OD600b	Bacteriocin peptide concn inc:				Antimicrobial activity as assessed by:
		ng/ml		ng/mg CDW		ADT (mm)d	MPA ine:
		EntL50A	EntL50B	EntL50A	EntL50B		BU/ml	BU/mg CDW
0	1.9	ND	ND	ND	ND	ND	ND	ND
24	4.0	3.3	20.5	1.6	9.7	7.9 s	40	20
48	4.2	6.8	42.0	3.1	19.1	8.1 s	70	30
72	4.3	8.8	44.3	4.0	20.0	10.2 s	310	140
96	4.4	10.3	46.4	4.7	21.7	10.7 s	360	170
120	4.4	9.4	55.3	4.2	24.8	10.3 s	320	140
144	4.5	8.4	45.7	3.7	20.4	9.8 s	260	120
168	4.5	7.1	36.0	3.2	16.1	9.3 s	210	90

^aS. cerevisiae L50A-20(pYABD02) and L50B-4(pYABD03) were grown at a 1:1 ratio in SCGR broth at 30°C.

^bGene expression was induced at time zero.

^cEntL50A and EntL50B concentration calculated by an NCI-ELISA using specific polyclonal antibodies for EntL50A or EntL50B, and expressed as ng/ml and ng/mg CDW. ND, no bacteriocin detected.

^dAntimicrobial activity against P. damnosus CECT4797 as determined by an ADT. Inhibition zones are differentiated as in Table 3. ND, no inhibition zone detected using 50 μl of supernatant.

^eAntimicrobial activity of supernatant against P. damnosus CECT4797 as determined by an MPA. ND, no inhibition detected using 100 μl of supernatant.