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Development of a multi-gene expression system in Xanthophyllomyces dendrorhous
Kiyotaka Y Hara1, Toshihiko Morita2, Masao Mochizuki3
1Organization of Advanced Science and Technology, Kobe University, 1-1 Rokkodaicho Nada-ku, Kobe, 657-8501, Japan. kiyotaka.hara@dolphin.kobe-u.ac.jp.
Microbial Cell Factories
|December 5, 2014
Summary
Researchers developed a novel multiple gene expression system in red yeast Xanthophyllomyces dendrorhous to boost astaxanthin production. This system enhances the mevalonate pathway, increasing astaxanthin yield by 2.1-fold without affecting cell growth.
Area of Science:
- Biotechnology
- Microbial Engineering
- Metabolic Engineering
Background:
- Xanthophyllomyces dendrorhous is the sole yeast producing astaxanthin, a valuable antioxidant used across industries.
- Enhancing intracellular mevalonate supply is key for astaxanthin production, but requires genetic engineering due to mevalonate's instability and cost.
- A robust system for simultaneous multiple gene expression is lacking in X. dendrorhous.
Purpose of the Study:
- To develop a multiple gene expression system for X. dendrorhous.
- To enhance the mevalonate synthetic pathway for increased astaxanthin production.
- To evaluate the impact of simultaneous overexpression of mevalonate pathway genes on astaxanthin yield.
Main Methods:
- Developed a plasmid-based system with strong promoters (actin, alcohol dehydrogenase, triose-phosphate isomerase) and terminators for X. dendrorhous.
- Overexpressed three key mevalonate synthetic pathway genes: acetoacetyl-CoA thiolase, HMG-CoA synthase, and HMG-CoA reductase.
- Combined overexpression of mevalonate pathway genes with genes involved in beta-carotene and astaxanthin synthesis.
Main Results:
- Achieved simultaneous overexpression of three mevalonate synthetic pathway genes using the developed system.
- Demonstrated a significant increase in astaxanthin production in the triple-overexpressing strain compared to single-gene overexpression strains.
- Observed a synergistic effect when combining mevalonate pathway enhancement with beta-carotene and astaxanthin synthesis genes, resulting in a 2.1-fold increase in astaxanthin production without compromising cell growth.
Conclusions:
- Successfully established a system for strong, multiple gene overexpression in X. dendrorhous.
- Enhanced mevalonate biosynthesis significantly increased astaxanthin production.
- The developed system holds potential for improving industrial isoprenoid production in X. dendrorhous by engineering platform strains.
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