Strain | Genotype | Source | Nanobody B |
---|
Titers (g l−1) | % Δ−16 Da variant |
---|
CBS7435MutS
|
aox1
| [7] | 2.4 ± 0.7 | 0.90 ± 0.17 |
CBS7435MutSΔOAHS |
aox1 met25
| This study | 2.1 ± 0.2 | 0 |
- Nanobody B was produced in the CBS7435MutS and in the CBS7435MutS strain with an inactivation of O-acetyl homoserine sulfhydrylase gene (CBS7435MutSΔOAHS) in shake flasks using complex media. Titers of Nanobody B were analyzed via a proteinA cleanup step followed by OD280 measurement. Relative abundance of the methoxine variant was analysed via RP-HPLC followed by total mass measurement by ESI-Q-TOF-MS. The methoxine variant of Nanobody B was only produced in the MutS strain and absent when O-acetyl homoserine sulfhydrylase was inactivated. Experiments were performed in triplicate