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Astaxanthin Overproduction Enhanced by Metabolomics-Guided Rational Metabolic Engineering in Synechococcus sp. PCC

Kousuke Ida1, Kenya Tanaka2,3,4, Yuichi Kato5

  • 1Kawasaki Frontience R&D Center, Toagosei Co., Ltd., 3-25-40 Tonomachi, Kawasaki, Kanagawa 210-0821, Japan.

ACS Synthetic Biology
|November 10, 2025
PubMed
Summary

Metabolic engineering of Synechococcus sp. PCC 7002 enhanced astaxanthin production. Overexpressing transketolase (TKT) increased astaxanthin content by 66%, demonstrating a metabolomics-guided approach for microbial production.

Keywords:
astaxanthincarotenoidcyanobacteriadesign-build-test-learn (DBTL)metabolomicstransketolase

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Area of Science:

  • Biotechnology and Synthetic Biology
  • Microbial Engineering
  • Metabolomics

Background:

  • Astaxanthin, a valuable antioxidant pigment, is primarily produced by *Haematococcus pluvialis*, but its cultivation is slow and prone to contamination.
  • *Synechococcus* sp. PCC 7002 offers a robust and fast-growing alternative for astaxanthin production, yet metabolic engineering strategies are underdeveloped.

Purpose of the Study:

  • To identify and address metabolic bottlenecks limiting astaxanthin production in *Synechococcus* sp. PCC 7002.
  • To enhance astaxanthin yield and productivity using a metabolomics-guided engineering approach.

Main Methods:

  • Engineered a base strain of *Synechococcus* sp. PCC 7002 with astaxanthin biosynthesis genes (*crtZ*, *crtW*).
  • Utilized metabolomics to identify rate-limiting enzymes, specifically transketolase (TKT) and IspG.
  • Overexpressed candidate genes (*tkt*, *ispG*) and optimized culture conditions (medium, light, temperature) to improve astaxanthin production.

Main Results:

  • The base strain produced 6.2 mg/g dry cell weight (DCW) of astaxanthin.
  • Metabolome analysis indicated bottlenecks at TKT and IspG, confirmed by reduced substrate accumulation upon gene overexpression.
  • Overexpression of *tkt* increased astaxanthin content to 10.3 mg/g-DCW (a 66% improvement), while *ispG* overexpression showed no significant effect. Optimized conditions yielded 7.5 mg/L/day productivity.

Conclusions:

  • A metabolomics-driven Design-Build-Test-Learn (DBTL) strategy effectively identified and relieved metabolic bottlenecks in *Synechococcus* sp. PCC 7002.
  • Targeted metabolic engineering, specifically TKT overexpression, significantly enhanced astaxanthin accumulation, paving the way for efficient microbial production.