Optimisation of MTGase production in Pichia pastoris under controlled growth rates and oxygen conditions
Acta Alimentaria, cilt.54, sa.4, ss.708-722, 2025 (SCI-Expanded, Scopus)
- Yayın Türü: Makale / Tam Makale
- Cilt numarası: 54 Sayı: 4
- Basım Tarihi: 2025
- Doi Numarası: 10.1556/066.2025.00214
- Dergi Adı: Acta Alimentaria
- Derginin Tarandığı İndeksler: Science Citation Index Expanded (SCI-EXPANDED), Scopus, BIOSIS
- Sayfa Sayıları: ss.708-722
- Anahtar Kelimeler: bioprocess, constant growth rate, hypoxia, MTGase, normoxia, Pichia pastoris
- Akdeniz Üniversitesi Adresli: Evet
Özet
In this study, microbial transglutaminase (MTGase) enzyme was produced recombinantly in Pichia pastoris with the constitutive GAP promoter under different growth rate-dependent normoxic and ethanol-dependent hypoxic conditions. The fermentations were performed at 20 °C temperature and the pH of 7. It was observed that the maximum enzyme production was 6236.3 U L−1 at the lowest specific growth rate 0.015 μ, in the normoxia fed-batch culture. While there was no statistically significant difference in wet cell weight at different growth rates under normoxic conditions, the highest total protein (307.7 mg L−1) was obtained at 0.015 µ. Furthermore, MTGase production was tested under hypoxic conditions with low oxygen demand by maintaining the ethanol concentration at 1% (v/v); however, unexpectedly, hypoxic conditions were not achieved at high pH and low temperature. It was concluded that hypoxia strategy is not suitable for producing MTGase in P. pastoris. This is the first study on optimisation of different growth rates and hypoxic conditions for MTGase production under the control of GAP promoter. In this study, the fermentation parameters were systematically optimised to increase the yield of MTGase enzyme.