In biofuel strain engineering, increasing gene copy number is often used to boost pathway flux and improve product titers. In theory, higher enzyme abundance increases conversion efficiency. In practice, overexpression can impose metabolic burden thus slowing growth and reducing overall space-time-yield.

At Aminoverse, we address this by identifying the true bottleneck in the pathway. Instead of adding more gene copies, we focus on weakest-link enzymes and improve catalytic performance through enzyme engineering. Key focus areas include improving catalytic efficiency, specificity and especially reducing kM to enhance substrate affinity, enabling high flux with fewer expressed copies and improved cellular fitness.

Example case study (biofuel application):

• Identified rate-limiting enzyme as primary pathway bottleneck
• Engineered enzyme to reduce kM approximately 5-fold, improving substrate affinity

• Reduced required gene copy number from 32 copies to 6, lowering metabolic burden and
improving growth
• Achieved 5% increase in product titer through improved pathway efficiency and faster strain
growth, increasing customer’s profitability by millions (ROI < 1 year)