Recombinant Polynucleotide Sequence for Astaxanthin Isomer Control

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Solution Overview

Problem

Current methods for astaxanthin biosynthesis face challenges such as low production yields, incorrect isomer formation, and susceptibility to oxidation, limiting their commercial and pharmaceutical applications due to differences in molecular properties between synthetic and natural astaxanthin.

Innovation Solution

A recombinant polynucleotide sequence comprising six gene cassettes, including those encoding geranylgeranyl pyrophosphate synthase, 3-hydroxy-3-methylglutaryl-coenzyme A reductase, phytoene desaturase, and β-carotene hydroxylase and ketolase, is introduced into Kluyveromyces marxianus to produce 3S, 3'S or 3R, 3'R astaxanthin, along with a vector and host cell, enabling efficient biosynthesis of astaxanthin and its derivatives.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Productivity

If chemical synthesis is used to produce astaxanthin, then production cost and availability are improved, but antioxidant activity and safety are worsened due to incorrect isomer formation and susceptibility to oxidation

Engineering Contradiction:
Improveproduction availabilityVSAvoidantioxidant activity
Core Design Contradiction:
ProductivityVSReliability

Solution Approach 1:

The patent uses genetically engineered microorganisms (yeast or bacteria) as living factories to produce astaxanthin through metabolic engineering. The microorganisms express carotenoid biosynthesis genes (crtE, crtI, crtY, crtZ, crtW) to synthesize astaxanthin with correct natural isomers (all-trans configuration and 3S,3'S stereoisomers), serving as an intermediary between chemical synthesis and natural extraction to provide both availability and biological activity

Inventive Principle:
Principle #24Intermediary (Mediator)

Solution Approach 2:

The patent optimizes multiple parameters including gene expression levels, cultural conditions, and metabolic flux to control the production of correct astaxanthin isomers. By adjusting these parameters, the system achieves high-yield production of biologically active all-trans astaxanthin while preventing formation of inactive cis-isomers

Inventive Principle:
Principle #35Parameter changes

2Reliability

If natural astaxanthin is extracted from algae or seafood, then correct isomer formation and antioxidant activity are improved, but production cost and availability are worsened

Engineering Contradiction:
Improveantioxidant activityVSAvoidproduction availability
Core Design Contradiction:
ReliabilityVSProductivity

Solution Approach 1:

The patent creates genetically engineered microorganisms that copy the carotenoid biosynthesis pathway from natural sources (algae, plants) into microbial hosts. By transferring and expressing the complete carotenoid gene cluster (crtE, crtI, crtY, crtZ, crtW) in yeast or bacteria, the system reproduces natural astaxanthin production with correct isomer formation while enabling scalable industrial fermentation

Inventive Principle:
Principle #26Copying

Solution Approach 2:

The patent divides the complex carotenoid biosynthesis pathway into discrete functional gene modules (crtE for GGPP synthesis, crtI for lycopene cyclization, crtY for hydroxylation, crtZ for ketolation, crtW for sterol C5-desaturase activity) that can be independently optimized and expressed in the microbial host to achieve controlled production

Inventive Principle:
Principle #1Segmentation

3Reliability

If all-trans astaxanthin is produced, then absorption and antioxidant activity are improved, but susceptibility to oxidation is worsened compared to esterified forms

Engineering Contradiction:
Improveantioxidant activityVSAvoidoxidation resistance
Core Design Contradiction:
ReliabilityVSStability of the object's composition

Solution Approach 1:

The patent employs dynamic metabolic control where the microorganism's metabolic state responds to environmental conditions. The system can dynamically adjust between producing free astaxanthin (high activity) and esterified astaxanthin (high stability) based on cultural conditions, allowing optimization for different application requirements

Inventive Principle:
Principle #15Dynamics

Applied Scientific Principles

This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.

Function Achieved in This Case

This approach enhances astaxanthin production, ensures correct isomer formation, and improves the antioxidant properties of the product, making it more suitable for commercial and pharmaceutical use by producing astaxanthin with higher antioxidant efficacy and stability.

Implementation Method 1

A recombinant polynucleotide sequence comprising six gene cassettes, including those encoding geranylgeranyl pyrophosphate synthase, 3-hydroxy-3-methylglutaryl-coenzyme A reductase, phytoene desaturase, and β-carotene hydroxylase and ketolase, is introduced into Kluyveromyces marxianus to produce 3S, 3'S or 3R, 3'R astaxanthin

Methodology Applied
Scientific EffectBiosynthesis:

Data Source

PatentEP3460044B1Recombinant polynucleotide sequence for producing astaxanthin and uses thereof
Publication Date: 2024.01.24 ACAD SINICA
  • EP3460044B1 patent drawingFigure 1
  • EP3460044B1 patent drawingFigure 2a
  • EP3460044B1 patent drawingFigure 2b

AI summary

Disclosed herein are recombinant polynucleotide sequences, vectors, host cells and methods for producing astaxanthin. The recombinant polynucleotide sequence is designed to provide a higher level of astaxanthin precursors via a shorter metabolic pathway, and thereby attains higher level of end products (e.g., astaxanthin) with desired stereoisomeric form and/or esterified form.