US 12,338,480 B2
Aconitic acid exporter (aexA) increases organic acid production in aspergillus
Shuang Deng, Richland, WA (US); Jon K. Magnuson, Richland, WA (US); Joonhoon Kim, Berkeley, CA (US); Kyle R. Pomraning, Richland, WA (US); Ziyu Dai, Richland, WA (US); and Beth A. Hofstad, Richland, WA (US)
Assigned to Battelle Memorial Institute, Richland, WA (US)
Filed by Battelle Memorial Institute, Richland, WA (US)
Filed on Oct. 24, 2023, as Appl. No. 18/493,590.
Application 18/493,590 is a division of application No. 17/347,109, filed on Jun. 14, 2021, granted, now 11,873,523.
Claims priority of provisional application 63/039,241, filed on Jun. 15, 2020.
Prior Publication US 2024/0076700 A1, Mar. 7, 2024
Int. Cl. C12P 7/46 (2006.01); C12N 1/14 (2006.01); C12R 1/69 (2006.01)
CPC C12P 7/46 (2013.01) [C12N 1/145 (2021.05); C12R 2001/69 (2021.05)] 16 Claims
OG exemplary drawing
 
1. A method of producing 3-hydroxypropionic acid (3-HP), comprising: culturing an isolated recombinant Aspergillus fungus in Riscaldati medium, or modified Riscaldati medium under conditions that permit the fungus to produce 3-HP, thereby making 3-HP,
wherein the fungus comprises:
an exogenous nucleic acid molecule encoding an aconitic acid exporter (aexA) protein comprising at least 60% sequence identity to SEQ ID NO: 2, 3, 4, or 5, operably linked to an exogenous promoter, thereby overexpressing the aexA in the fungus;
an endogenous or exogenous nucleic acid molecule encoding aspartate 1-decarboxylase (panD);
an endogenous or exogenous nucleic acid molecule encoding β-alanine-pyruvate aminotransferase (BAPAT); and
an endogenous or exogenous nucleic acid molecule encoding 3-hydroxypropionate dehydrogenase (3-HPDH), and
wherein the Riscaldati medium comprises 100 g/L glucose, 0.11 g/L KH2PO4, 2.36 g/L (NH4)2SO4, 2.08 g/L MgSO4*7H2O, 0.074 g/L NaCl, 0.13 g/L CaCl2*2H2O, 0.0013 g/L ZnSO4*7H2O, 0.0055 g/L FeSO4*7H2O, 0.0002 g/L CuSO4*5H2O, and 0.0007 g/L MnCl2*4H2O, or
wherein the modified Riscaldati medium comprises 100 g/L glucose, 0.11 g/L KH2PO4, 2.36 g/L (NH4)2SO4, 2.08 g/L MgSO4*7H2O, 0.074 g/L NaCl, 0.13 g/L CaCl2*2H2O, 20 times 0.0013 g/L ZnSO4*7H2O, 20 times 0.0055 g/L FeSO4*7H2O, 20 times 0.0002 g/L CuSO4*5H2O, and 20 times 0.0007 g/L MnCl2*4H2O.