Cofactor engineering improved CALB production in Pichia pastoris through heterologous expression of NADH oxidase and adenylate kinase

• DOI: 10.1371/journal.pone.0181370
• Publication Date: 2017-07-17T17:47:22Z
• Authors: Charumathi Jayach...; Balakumaran Palan...; Meenakshisundaram...
• Keywords: Energy charge; Heterologous expression

The cofactor engineering strategy can relieve the metabolic stress induced by expression of recombinant protein in cellular metabolism related to and energy reactions. To study effect regeneration on expression, NADH oxidase (noxE) was engineered P. pastoris expressing lipase B (GSCALB). Expression noxE (GSCALBNOX) increased NAD+ levels 85% with a concomitant reduction NADH/NAD+ ratio 67% compared GSCALB. change redox level positively influenced methanol uptake rate made 34% augment CALB activity. decline (44%) had lowered adenylate charge (AEC) ATP GSCALBNOX. In order regenerate GSCALBNOX, kinase (ADK1) gene from S. cerevisiae S288c co-expressed. ADK1 showed remarkable increase AEC co-expression both genes synergistically improved This shows importance maintenance homeostasis during pastoris.