Synthesis of Secretory Proteins in Yarrowia lipolytica: Effect of Combined Stress Factors and Metabolic Load
Abstract
:1. Introduction
2. Results
2.1. Cell Growth and Substrate Consumption Kinetics by the Differentially Burdened Strains under Extrinsic Stress Conditions
2.2. Heterologous Gene Expression under the Applied Stress Conditions
2.3. Metabolic Activity and Morphology of the Cells
2.4. Secretion of Active rs-Prots under High and Low OA
2.5. Gene Expression vs. rs-Prot Synthesis
3. Discussion
4. Materials and Methods
4.1. Strains and Basic Culture Conditions
4.2. Main Cultures and Implementation of Stress Factors
4.3. Gene Expression Analysis
4.4. Analytical Methods
4.4.1. Flow Cytometry
4.4.2. Enzymatic Assays
4.4.3. Chromatography
4.4.4. Microscopy
4.4.5. Statistical Analysis
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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pH and OA Conditions | Strain | Growth Rate QgDCW [g L−1 h−1] | Glycerol Consumption Rate Qgly [g L−1 h−1] $ | Biomass Yield Yx/s [g g−1] $ | Max. Biomass [g L−1] | Specific Secretion of rs-Prots [FU gDCW−1 or AU gDCW−1] | Total Secretion of Rs-Prots [FU or AU] |
---|---|---|---|---|---|---|---|
pH 3/kLa 28 | LB | 0.085 ± 0.002 d | 0.16 ± 0.01 | 0.54 ± 0.01 | 3.37 ± 0.03 f | N.D.—irreversible degradation of rs-Prots under pH 3 | |
HB | 0.11 ± 0.01 d | 0.28 ± 0.01 | 0.39 ± 0.02 | 4.1 ± 0.1 f | |||
control | 0.080 ± 0.002 d | N.A. | N.A. | 3.60 ± 0.01 f | N.D. | N.D. | |
pH 3/kLa 110 | LB | 0.26 ± 0.02 b | 0.74 ± 0.01 | 0.36 ± 0.03 | 8.6 ± 0.5 bc | N.D.—irreversible degradation of rs-Prots under pH 3 | |
HB | 0.275 ± 0.004 b | 0.65 ± 0.03 | 0.43 ± 0.02 | 8.4 ± 0.3 c | |||
control | 0.28 ± 0.03 b | N.A. | N.A. | 7.3 ± 0.6 cd | N.D. | N.D. | |
pH 7/kLa 28 | LB | 0.118 ± 0.002 d | 0.21 ± 0.01 | 0.57 ± 0.01 | 6.0 ± 0.2 e | 132 ± 27 | 459 ± 96 |
HB | 0.18 ± 0.01 c | 0.37 ± 0.02 | 0.50 ± 0.01 | 9.73 ± 0.03 b | 0.0029 ± 0.0004 | 0.015 ± 0.001 | |
control | 0.113 ± 0.004 d | N.A. | N.A. | 6.1 ± 0.2 de | N.D. | N.D. | |
pH 7/kLa 110 | LB | 0.46 ± 0.01 a | 0.83 ± 0.01 a | 0.55 ± 0.01 | 11.6 ± 0.1 a | 597 ± 83 | 6944 ± 1009 |
HB | 0.25 ± 0.02 b | 0.82 ± 0.01 a | 0.31 ± 0.02 | 7.5 ± 0.5 c | 0.0036 ± 0.0003 | 0.024 ± 0.002 | |
control | 0.50 ± 0.02 a | N.A. | N.A. | 12.6 ± 0.5 a | N.D. | N.D. |
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Gorczyca, M.; Kaźmierczak, J.; Fickers, P.; Celińska, E. Synthesis of Secretory Proteins in Yarrowia lipolytica: Effect of Combined Stress Factors and Metabolic Load. Int. J. Mol. Sci. 2022, 23, 3602. https://doi.org/10.3390/ijms23073602
Gorczyca M, Kaźmierczak J, Fickers P, Celińska E. Synthesis of Secretory Proteins in Yarrowia lipolytica: Effect of Combined Stress Factors and Metabolic Load. International Journal of Molecular Sciences. 2022; 23(7):3602. https://doi.org/10.3390/ijms23073602
Chicago/Turabian StyleGorczyca, Maria, Jan Kaźmierczak, Patrick Fickers, and Ewelina Celińska. 2022. "Synthesis of Secretory Proteins in Yarrowia lipolytica: Effect of Combined Stress Factors and Metabolic Load" International Journal of Molecular Sciences 23, no. 7: 3602. https://doi.org/10.3390/ijms23073602