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Expression of Recombinant Proteins in the Methylotrophic Yeast Pichia pastoris
Published on: February 25, 2010
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Efficient microbial production of stylopine using a Pichia pastoris expression system
Kentaro Hori1, Shunsuke Okano1, Fumihiko Sato1
1Division of Integrated Life Science, Graduate School of Biostudies, Kyoto University, Kitashirakawa, Sakyo, Kyoto 606-8502, Japan.
Scientific Reports
|March 1, 2016
Summary
Microbial production of stylopine from (S)-reticuline was achieved using a consolidated or co-culture system in Pichia pastoris. The co-culture system demonstrated superior long-term efficiency for stylopine biosynthesis.
Area of Science:
- Biotechnology
- Metabolic Engineering
- Synthetic Biology
Background:
- Stylopine is a protoberberine alkaloid with potential biological activities.
- Microbial production of (S)-reticuline has been previously established.
- Efficient biosynthesis of stylopine is crucial for exploring its applications.
Purpose of the Study:
- To develop microbial production systems for stylopine from (S)-reticuline.
- To compare the efficiency of a consolidated system versus a co-culture system for stylopine biosynthesis.
- To optimize stylopine production using Pichia pastoris.
Main Methods:
- Expression of berberine bridge enzyme (CYP719A5) and stylopine synthase (CYP719A2) in Pichia pastoris.
- Comparison of a single-cell consolidated system with a three-cell-line co-culture system.
- Evaluation of substrate-feeding strategies and conversion efficiency over successive cultures.
Main Results:
- Both consolidated and co-culture systems successfully converted reticuline to stylopine.
- The consolidated system showed higher initial conversion rates.
- The co-culture system maintained stable conversion efficiency during successive feedings, yielding over 150 nmoles of stylopine from 750 nmoles of reticuline.
Conclusions:
- The co-culture system is more robust for sustained, high-yield microbial production of stylopine.
- Pichia pastoris is a suitable host for engineering stylopine biosynthesis.
- Further optimization of microbial cell factories can enhance alkaloid production.
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