Dark-Active Cryptochrome Mutations for Algal Growth in Darkness
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Solution Overview
Problem
Algae typically grow poorly in the dark, limiting their industrial applications, as they are either unable to grow heterotrophically or have slow growth rates and low maximum densities in dark conditions.
Innovation Solution
Introduction of a dark active cryptochrome into algae, specifically through mutations in the flavin adenine dinucleotide-binding domain, such as substituting the glycine residue at position 8 with a basic amino acid, to maintain activation and enhance growth in the dark.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If algae are cultured in the dark using heterotrophic growth, then energy consumption is reduced and industrial scalability is improved, but growth rate and maximum algal density become slow and low
Solution Approach 1:
The invention changes the functional state of the cryptochrome protein by mutating specific amino acid residues (e.g., G380R substitution in Arabidopsis CRY1) to create a constitutively active form that functions in the dark. This parameter change in protein activation state enables heterotrophic growth capability and improves growth rate without requiring light conditions
Solution Approach 2:
The invention replaces the light-dependent activation mechanism (photoreception) with a constitutive activation mechanism through protein engineering. The mutated cryptochrome protein maintains active conformation without requiring blue light activation, substituting the optical activation mechanism with a stable structural configuration that enables dark activity
2Productivity
If algae are cultured in the dark, then scalability and industrial application potential are improved, but growth rate and maximum density remain low
Solution Approach 1:
The mutation of cryptochrome amino acid residues creates a constitutively active protein form that maintains high growth rates in the dark across multiple subcultures. This parameter change in protein activation state prevents the typical decline in growth rate that occurs after subculture in dark conditions
Solution Approach 2:
The constitutively active cryptochrome protein maintains continuous heterotrophic growth capability through multiple subculture cycles in the dark. The useful action of dark growth is sustained continuously without the typical decline that occurs after subculture, enabling long-term industrial cultivation
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
Algae with dark active cryptochrome exhibit significantly improved growth rates and higher maximum densities in the dark, making them more suitable for industrial use.
Implementation Method 1
The dark active cryptochrome has an amino acid sequence having a mutation in a flavin adenine dinucleotide-binding domain compared to the amino acid sequence of a dark inactive cryptochrome
Data Source
AI summary
An alga having a dark active cryptochrome whose activation is maintained in the dark. Also, a method for improving growth of an alga in the dark including introducing a polynucleotide encoding a dark active cryptochrome into an alga. Also, a method for producing an alga with improved growth in the dark having an improved growth rate including introducing a polynucleotide encoding a dark active cryptochrome into an alga.