5-Ribonucleotide V-IV-B

Technical Specifications

Pathway Edits

• ▸LeADSS adenylosuccinate synthase overexpression under strong gpd promoter for de-novo IMP elevation feeding all three 5′-ribonucleotide branches
• ▸LeIMPDH2 inosine monophosphate dehydrogenase isoform 2 upregulation for GMP branch augmentation alongside IMP accumulation
• ▸LeAK1 adenylate kinase 1 overexpression for AMP/ADP/ATP equilibration, maintaining free AMP concentration as third umami 5′-ribonucleotide co-agonist
• ▸LeAMPD AMP deaminase partial knockdown (45%) to preserve AMP from conversion to IMP, maintaining all three 5′-nucleotide species simultaneously
• ▸LeRNase T1 upregulation for endogenous RNA hydrolysis releasing purine nucleotides in post-harvest drying phase of shiitake production
• ▸LePNP purine nucleoside phosphorylase activity optimization for balanced inosine/guanosine/adenosine salvage supporting all three 5′-nucleotide pools
• ▸LeNT5′ 5′-nucleotidase balanced expression for free nucleotide fraction maintenance from monophosphate pools
• ▸LeADA adenosine deaminase partial suppression (35%) to slow AMP-to-IMP conversion, preserving AMP as independent umami 5′-nucleotide
• ▸LeHPRT hypoxanthine-guanine phosphoribosyltransferase upregulation for hypoxanthine/guanine salvage to IMP/GMP complementing de-novo pathway

Key Interventions