TABLE 4.
Cosynthesis assays in liquid SA mediuma
| Cosynthesis assaya | Strain (gene mutated)b |
Clavulanic acid bioassay resultc | |
|---|---|---|---|
| Donor | Receptor | ||
| 1 | S. clavuligerus ΔoppA2::acc (oppA2) | S. clavuligerus pyc::aph (ceaS) | ++++ |
| 2 | S. clavuligerus ΔoppA2::acc (oppA2) | S. clavuligerus RFL35 (bls2) | ++++ |
| 3 | S. clavuligerus ΔoppA2::acc (oppA2) | S. clavuligerus orf14::aph | −−−−− |
| 4 | S. clavuligerus ΔoppA2::acc (oppA2) | S. clavuligerus cyp::aph (cyp) | −−−−− |
| 5 | S. clavuligerus ΔoppA2::acc (oppA2) | S. clavuligerus oppA1::aph (oppA1) | −−−−− |
| 6 | S. clavuligerus cyp::aph (cyp) | S. clavuligerus pyc::aph (ceaS) | −−−−− |
| 7 | S. clavuligerus cyp::aph (cyp) | S. clavuligerus RFL35 (bls2) | −−−−− |
| 8 | S. clavuligerus pyc::aph (ceaS) | S. clavuligerus ΔoppA2::acc (oppA2) | −−−−− |
| 9 | S. clavuligerus RFL35 (bls2) | S. clavuligerus ΔoppA2::acc (oppA2) | −−−−− |
a
Assays 1 to 5 were also carried out using mixed cultures of both strains. The complementation results in the bioassay were identical.
b
Donor strains were from spent sterile supernatants; receptor strains were from growing cultures.
c
++++, positive cosynthesis; −−−−−, negative cosynthesis. CA bioassays of each separated strain were always negative. None of the strains shown in this table produces clavulanic acid by itself. All the strains produced cephamycin C.