Diatom Ovothiol Biosynthesis via Enzymatic Engineering

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Solution Overview

Problem

Current methods for producing ovothiols are cumbersome, expensive, and environmentally unfriendly, with chemical synthesis procedures being time-consuming and sea urchins being an unsustainable source for large-scale production.

Innovation Solution

Development of a protocol for enzymatic engineering of the diatom species Phaeodactylum tricornutum to overexpress the biosynthetic enzyme for ovothiol production, using bacterial conjugation or biolistic methods to introduce the 5-histidylcysteine sulfoxide synthase OvoA enzyme, enabling eco-sustainable and cost-effective production of ovothiols.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Productivity

If chemical synthesis procedures are used to produce ovothiols, then the production process can be controlled, but the process becomes time-consuming and expensive

Engineering Contradiction:
Improveovothiol production efficiencyVSAvoidsynthesis time
Core Design Contradiction:
ProductivityVSLoss of time

Solution Approach 1:

The patent uses genetically modified diatoms that autonomously produce ovothiols through their own metabolic pathways. The microalgae cells serve as living factories that self-regulate the biosynthesis process, eliminating the need for manual chemical synthesis steps and significantly reducing production time while maintaining control over the production process.

Inventive Principle:
Principle #25Self-service

Solution Approach 2:

The patent replaces mechanical/chemical synthesis systems with a biological system. Instead of using chemical reagents and multi-step synthesis protocols, the invention employs genetically engineered microorganisms that naturally produce ovothiols through enzymatic reactions, thereby reducing both time and complexity of the production process.

Inventive Principle:
Principle #28Mechanics substitution (Replace mechanical system)

2Reliability

If sea urchins are used as a source for ovothiol extraction, then natural ovothiols can be obtained, but the method is unsustainable for large-scale production

Engineering Contradiction:
Improveovothiol source reliabilityVSAvoidscale of production
Core Design Contradiction:
ReliabilityVSProductivity

Solution Approach 1:

The patent creates a copy of the ovothiol production capability by transferring the biosynthetic genes into diatom cells. Instead of extracting ovothiols from sea urchins, the invention replicates the production mechanism in a more scalable host organism, maintaining the reliability of natural ovothiol production while enabling large-scale cultivation through controlled diatom growth.

Inventive Principle:
Principle #26Copying

Solution Approach 2:

The patent changes the biological parameters of the production system by switching from sea urchin extraction to diatom cultivation. This parameter change includes using organisms with faster reproduction rates and higher biomass yield, thereby increasing productivity and scalability while maintaining the quality and reliability of natural ovothiol production.

Inventive Principle:
Principle #35Parameter changes

3Object-affected harmful factors

If enzymatic engineering of diatoms is implemented, then eco-sustainable production is achieved, but the genetic modification process adds complexity

Engineering Contradiction:
Improvetoxic side compoundsVSAvoidgenetic engineering complexity
Core Design Contradiction:
Object-affected harmful factorsVSDevice complexity

Solution Approach 1:

The patent extracts and isolates the specific biosynthetic genes responsible for ovothiol production and transfers them into diatoms. By taking out only the essential genetic elements needed for ovothiol synthesis, the invention achieves eco-sustainable production without requiring complex whole-organism genetic改造工程, thereby reducing overall system complexity while eliminating toxic side compounds associated with chemical synthesis.

Inventive Principle:
Principle #2Taking out (Extraction)

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 results in higher productivity and scalability of ovothiol production, avoiding toxic side compounds and preserving natural populations, making ovothiols more accessible for pharmaceutical, nutraceutical, and cosmeceutical applications.

Implementation Method 1

The key enzyme involved in ovothiol biosynthesis is a bifunctional enzyme, which catalyzes the formation of the oxidative C-S bond, which results in the net sulfur transfer from cysteine to position 5 of histidine. In detail, the enzyme 5-histidylcysteine sulfoxide synthase (OvoA) catalyzes the formation of the 5-histidyl-cysteine sulfoxide conjugate from cysteine and histidine

Methodology Applied
Scientific EffectEnzyme catalysis: Enzyme

Implementation Method 2

Subsequently, a pyridoxal phosphate (PLP)-dependent lyase (OvoB) cleaves this intermediate product to generate 5-thiohistidine

Methodology Applied
Scientific EffectEnzyme catalysis: Enzyme

Implementation Method 3

Then, OvoA catalyzes the methylation at the imidazole ring to produce ovothiol A

Methodology Applied
Scientific EffectEnzyme catalysis: Enzyme

Data Source

PatentEP4166658A1Diatom-based genetic engineering system methodology for the eco-sustainable production of ovothiols
Publication Date: 2023.04.19 STAZIONE ZOOLOGICA ANTON DOHRN
  • EP4166658A1 patent drawingFigure 1~2
  • EP4166658A1 patent drawingFigure 3~4
  • EP4166658A1 patent drawingFigure 5A~5G

AI summary

The present invention relates to the field of biotechnology, in particular it relates to the set-up of a protocol of enzymatic engineering of the diatom species Phaeodactylum tricornutum to overexpress the biosynthetic enzyme leading to ovothiol production. This protocol is eco-sustainable because it uses cells and nutrients for biosynthesis and does not produce toxic side-compounds. Microalgal biomass moreover can be exploited for additional uses after ovothiol extraction, making the production process more cost-effective. The production of ovothiols is relevant for the pharmaceutical, nutraceutical, and cosmeceutical sectors.