DGAT Genes With Pleckstrin Homology Domains For Algal Lipid Production
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Solution Overview
Problem
Current methods for boosting triglyceride (TAG) production in algae, such as alternating nutrient conditions or heterotrophic growth, are costly and prone to contamination, and there is limited understanding of the subcellular localization of recombinant diacylglycerol acyltransferase (DGAT) enzymes, which are crucial for TAG synthesis.
Innovation Solution
Incorporating Pleckstrin Homology (PH) domains into DGAT genes in recombinant microorganisms, specifically in algal species, to enhance lipid production by improving the subcellular localization and activity of DGAT enzymes, thereby increasing TAG synthesis.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Quantity of substance
If algae are grown under nitrogen limitation to increase TAG production, then TAG concentration is improved, but overall lipid productivity deteriorates due to limited biomass accumulation
Solution Approach 1:
The invention changes the cultural parameters from nitrogen limitation to nitrogen sufficiency combined with heterotrophic carbon limitation. This parameter shift allows simultaneous achievement of high biomass accumulation and high TAG content, resolving the contradiction between TAG concentration and overall productivity
2Productivity
If algae are grown heterotrophically with extra organic carbon to boost TAG production, then TAG production is improved, but contamination risk worsens due to stimulation of exogenous bacteria or fungi
Solution Approach 1:
The invention changes the carbon source parameter from readily fermentable sugars to non-fermentable carbon sources such as C4 sugars or C6-C12 carboxylic acids. This modification maintains heterotrophic TAG production while eliminating the contamination risk associated with traditional sugar-based heterotrophic growth
3Productivity
If two-step nutrient alternating process is used to boost TAG production, then TAG production is improved, but production cost worsens due to long growth periods with little TAG production
Solution Approach 1:
The invention enables continuous production of high-value TAG throughout the entire growth cycle by maintaining heterotrophic conditions with carbon limitation. This eliminates the need for alternating between nutrient-rich and nutrient-limited phases, achieving continuous useful action rather than intermittent production
4Productivity
If recombinant DGAT enzymes are introduced to enhance TAG synthesis, then TAG synthesis capability is improved, but subcellular localization understanding worsens due to limited knowledge of enzyme localization
Solution Approach 1:
The invention uses PH domains as intermediary localization signals to direct DGAT enzymes to specific subcellular locations (endoplasmic reticulum or plasma membrane). This intermediary mechanism resolves the localization uncertainty by providing a known targeting sequence that mediates enzyme placement
Data Source
AI summary
The present invention provides novel diacylglycerol acyltransferase (DGAT) genes comprising Pleckstrin Homology (PH) domains. The present invention also provides for recombinant cells, such as algae, transformed with acyltransferase genes, such as DGAT, comprising PH domains, and methods of using such recombinant cells to produce increased triglyceride levels.