An Engineered Yeast Efficiently Secreting Penicillin
Figure 6
H. polymorpha cells produce and secrete PenG: LC MS/MS demonstration of PEN production.
Strain HpPen4 was grown in a glucose/methanol-limited continuous culture in the presence of 1 mM AAA and 1 mM PAA. Spent medium was analyzed by LC-MS/MS. The LC chromatograms show the presence of IPN (3.45 min; Fig. 6A) and PenG (11.76 min) and a PenG-related product (7.83 min) (Fig. 6C). Fig. 6B and D show the LC-MS/MS fragmentation patterns corresponding to IPN (Fig. 6B) and PenG (Fig. 6D). The characteristic MS/MS fragmentation pattern of pure IPN and PenG and their relative abundance are represented in the tables present in Fig 6A and C.