PART COLLECTION
Erythritol is a non-cariogenic sweetener. Currently, it is primarily produced through microbial fermentation using glucose as the main substrate. However, this method suffers from limitations, such as low yield and conversion rate. Glycerol, being an abundant and low-value renewable resource, will be utilized as substrate in our research to achieve dual-substrate fermentation for erythritol production. The project is divided into two cycles(Liu F etc, 2024; Thuy etc, 2024):
Cycle 1 - Glucose-based erythritol synthesis[Pathway 1]
pCDFDuet-PK-EPDH-PTase, BBa_25VEGQXC
Cycle 2 - Glycerol-based erythritol synthesis[Pathway 2]
pETDuet-GUT12-TPI1, BBa_25XX8745
Cycle 1 and Cycle 2 are complementary. Cycle 2 can utilize glycerol as a substrate to produce erythritol only if the proteins, when Cycle 1 are normally expressed. Furthermore, this project provides a solid foundation for future explorations of these techniques in glycerol reuse. The comprehensive and systematic approach outlined herein is critical to navigating the complex regulatory environment.
Figure 1.Synthesis pathway of erythritol
(Note: PK: phosphoketolase; EPDH: erythritol-4-phosphate dehydrogenase; PTase: phosphatase; GUT1: Encoding glycerol kinase; GUT2: Encoding glycerol-3-phosphate dehydrogenase; TPI1: Encoding triosephosphate isomerase)
The Part Library we constructed is as follows:
|
Number |
Name |
Part type |
Part type |
Part function |
Part diagram |
|
PK |
Basic Part |
Coding |
This enzyme catalyzes the cleavage of phosphoketoses (fructose-6-phosphate etc.) to generate smaller phosphorylated products |
|
|
|
EPDH |
Basic Part |
Coding |
It catalyzes the dehydrogenation of erythritol-4-phosphate (E4P), converting it into erythrulose-4-phosphate (E4PuP) |
|
|
|
PTase |
Basic Part |
Coding |
Phosphatase operates as an enzyme that removes phosphate groups from its substrates by breaking down phosphate monoesters which results in phosphate ions and free hydroxyl groups |
|
|
|
GUT1 |
Basic Part |
Coding |
Glycerol kinase catalyzes the conversion of glycerol into glycerol-3-phosphate |
|
|
|
GUT2 |
Basic Part |
Coding |
Glyceraldehyde-3-phosphate dehydrogenase (GAPD) is an NAD-dependent oxidoreductase with phosphorylating activity. |
|
|
|
TPI1 |
Basic Part |
Coding |
Triosephosphate isomerase (TPI) is a key enzyme in the glycolytic pathway, catalyzing the reversible isomerization between dihydroxyacetone phosphate (DHAP) and glyceraldehyde-3-phosphate (GAP) |
|
|
|
pCDFDuet |
Basic Part |
Plasmid_Backbone |
Co-transformation expression vector |
|
|
|
pETDuet |
Basic Part |
Plasmid_Backbone |
Co-transformation expression vector |
|
|
|
pCDFDuet-PK-EPDH-PTase |
Composite Part |
Plasmid |
Glucose-based erythritol synthesis |
|
|
|
pETDuet-GUT12-TPI1 |
Composite Part |
Plasmid |
Glycerol-based erythritol synthesis |
|
References
Liu F, Tian JT, Wang YT, Zhao L, Liu Z, Chen J, Wei LJ, Fickers P, Hua Q. Improving an Alternative Glycerol Catabolism Pathway in Yarrowia lipolytica to Enhance Erythritol Production. Yeast. 2024 Oct;41(10):605-614. doi: 10.1002/yea.3980. Epub 2024 Sep 11. PMID: 39262092.
Thuy Le H, Vu YT, Duong GH, Le TK, Dang MK, Pham DD, Pham NK, Sichaem J, Nguyen NH, Duong TH. Bio-Guided Isolation of Alpha-Glucosidase Inhibitory Compounds from Vietnamese Lichen Roccella Montagnei. Chem Biodivers. 2024 Jul;21(7):e202400438. doi: 10.1002/cbdv.202400438. Epub 2024 May 28. PMID: 38581153.
