Engineered Production of Isobutanol from Sugarcane Trash Hydrolysates in Pichia pastoris
Abstract
:1. Introduction
2. Materials and Methods
2.1. Yeast Strain, Media, and Transformation
2.2. Plasmid Construction
2.3. Quantification of Isobutanol and Other Metabolites in Engineered Strains
2.4. Determination of the Engineered Strains’ Specific Growth Rates
2.5. Sugarcane Trash Pretreatment and Enzymatic Hydrolysis
2.6. RNA Isolation and Transcript Quantification
3. Results and Discussion
3.1. Overexpression of Xylose Isomerase and Xylulokinase in P. pastoris Enables Cells to Grow in a Medium Containing Xylose as the Sole Carbon Source
3.2. Engineered Strains with Overexpression of Xylose Isomerase and Xylulokinase Produce Higher Titer of Isobutanol in a Medium Containing Xylose and Glucose as a Mixed Carbon Source
3.3. Preparation of Sugarcane Trash Hydrolysates
3.4. Isobutanol Production from Sugarcane Trash Hydrolysates
4. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Strain Name | Overexpressed Genes 1 | Specific Growth Rates in MGYH-glu (h−1) | Specific Growth Rates in MXYH(h−1) | References |
---|---|---|---|---|
KM71 | None | 0.07583 ± 0.00066 | 0.00070 ± 0.00008 | Invitrogen |
PPY0001 | PspXI | 0.07790 ± 0.00099 | 0.00077 ± 0.00005 | This paper |
PPY0002 | LpXI | 0.07730 ± 0.00108 | 0.00083 ± 0.00005 | This paper |
PPY0011 | PspXI, PpXKS | 0.05330 ± 0.00106 | 0.00267 ± 0.00005 | This paper |
PPY0012 | LpXI, PpXKS | 0.06050 ± 0.00128 | 0.00317 ± 0.00005 | This paper |
PPY0111 | PspXI, PpXKS, PpIlv2, PpIlv6 | 0.05747 ± 0.00094 | 0.00257 ± 0.00008 | This paper |
PPY0112 | LpXI, PpXKS, PpIlv2, PpIlv6 | 0.05900 ± 0.00179 | 0.00260 ± 0.00008 | This paper |
PPY0211 | PspXI, PpXKS, PpIlv5, PpIlv3, LlkivD, ScADH7 | 0.04963 ± 0.00264 | 0.00193 ± 0.00017 | This paper |
PPY0212 | LpXI, PpXKS, PpIlv5, PpIlv3, LlkivD, ScADH7 | 0.05813 ± 0.00133 | 0.00287 ± 0.00005 | This paper |
PPY0311 | PspXI, PpXKS, PpIlv2, PpIlv6, PpIlv5, PpIlv3, LlkivD, ScADH7 | 0.05097 ± 0.00301 | 0.00260 ± 0.00008 | This paper |
PPY0312 | LpXI, PpXKS, PpIlv2, PpIlv6, PpIlv5, PpIlv3, LlkivD, ScADH7 | 0.06153 ± 0.00111 | 0.00280 ± 0.00014 | This paper |
PPY0100 | PpIlv2, PpIlv6 | 0.07303 ± 0.00264 | 0.00067 ± 0.00005 | This paper |
PPY0200 | PpIlv5, PpIlv3, LlkivD, ScADH7 | 0.06573 ± 0.00146 | 0.00073 ± 0.00005 | This paper |
PPY0300 | PpIlv2, PpIlv6, PpIlv5, PpIlv3, LlkivD, ScADH7 | 0.06330 ± 0.00206 | 0.00083 ± 0.00005 | This paper |
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Bumrungtham, P.; Promdonkoy, P.; Prabmark, K.; Bunterngsook, B.; Boonyapakron, K.; Tanapongpipat, S.; Champreda, V.; Runguphan, W. Engineered Production of Isobutanol from Sugarcane Trash Hydrolysates in Pichia pastoris. J. Fungi 2022, 8, 767. https://doi.org/10.3390/jof8080767
Bumrungtham P, Promdonkoy P, Prabmark K, Bunterngsook B, Boonyapakron K, Tanapongpipat S, Champreda V, Runguphan W. Engineered Production of Isobutanol from Sugarcane Trash Hydrolysates in Pichia pastoris. Journal of Fungi. 2022; 8(8):767. https://doi.org/10.3390/jof8080767
Chicago/Turabian StyleBumrungtham, Pornsiri, Peerada Promdonkoy, Kanoknart Prabmark, Benjarat Bunterngsook, Katewadee Boonyapakron, Sutipa Tanapongpipat, Verawat Champreda, and Weerawat Runguphan. 2022. "Engineered Production of Isobutanol from Sugarcane Trash Hydrolysates in Pichia pastoris" Journal of Fungi 8, no. 8: 767. https://doi.org/10.3390/jof8080767