Pichia pastoris growth-coupled heme biosynthesis analysis using metabolic modelling

Area of Science:

• Biotechnology
• Metabolic Engineering
• Food Science

Background:

• Soy leghemoglobin is crucial for plant-based meat color and flavor.
• High-yield production of leghemoglobin and its heme component is essential for the food industry.

Purpose of the Study:

• To enhance the high-yield production of leghemoglobin and heme in Pichia pastoris.
• To identify optimal cultivation conditions and metabolic engineering strategies for increased heme production.

Main Methods:

• Cultivation of Pichia pastoris on glycerol and methanol.
• In-silico metabolic modeling for growth-coupled enzyme quantity analysis.
• Identification of gene up/down-regulation strategies for metabolic optimization.

Main Results:

• Glycerol cultivation showed potential for a 50% increase in uptake and production rates.
• Methanol cultivation approached theoretical maximums.
• Metabolic modeling suggested 33 reactions for upregulation and 66 for downregulation/deletion to optimize heme production.

Conclusions:

• Metabolic modeling provides effective strategies for improving heme production in Pichia pastoris.
• Optimized cultivation and gene regulation can significantly boost leghemoglobin yield for meat substitutes.