Optimizing Trilobatin Production via Screening and Modification of Glycosyltransferases
Yue Yang1,2, Yuhan Cheng2, Tao Bai1
1Jiaxing Synbiolab Biotechnology Co., Ltd., Jiaxing 314006, China.
Molecules (Basel, Switzerland)
|February 10, 2024
Summary
Researchers enhanced the production of trilobatin (TBL), a natural sweetener, using an engineered enzyme. This improved enzymatic precision and reduced by-products, paving the way for industrial-scale TBL synthesis.
Area of Science:
- Biotechnology
- Enzyme Engineering
- Natural Product Synthesis
Background:
- Trilobatin (TBL) is a natural dihydrochalcone sweetener from *Rubus suavissimus S. Lee* with health benefits.
- Current botanical extraction methods for TBL are not cost-effective or scalable.
- Existing enzymatic synthesis using ph-4'-OGT is limited by low efficiency and by-product formation.
Purpose of the Study:
- To improve the enzymatic synthesis of trilobatin (TBL).
- To enhance the catalytic efficiency and reduce by-products in TBL biosynthesis.
- To establish a foundation for the industrial-scale production of TBL.
Main Methods:
- Screened 17 *Md*ph-4'-OGT enzyme sequences for TBL synthesis.
- Optimized reaction conditions for the most effective enzyme variant, PT577.
- Utilized virtual screening to identify mutation sites and create engineered enzyme strains.
Main Results:
- Seven of the screened enzymes showed catalytic activity for TBL synthesis.
- PT577 achieved a 97.3% conversion yield in 3 hours, with optimized bioproduction reaching 163.3 mg/L.
- Engineered PT577 strains reduced by-products by up to 50%.
Conclusions:
- The study successfully enhanced enzymatic precision for TBL biosynthesis.
- Optimized enzyme variants and reaction conditions provide a robust foundation for industrial TBL production.
- Findings have broader implications for glycosyltransferase engineering and in silico analysis.
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