Biotechnology Bulletin ›› 2022, Vol. 38 ›› Issue (11): 227-237.doi: 10.13560/j.cnki.biotech.bull.1985.2022-0280

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Metabolic Engineering Modification of Aspergillus niger for the Production of D-glucaric Acid

GUO Yu-fei(), YAN Rong-mei, ZHANG Xiao-ru, CAO Wei, LIU Hao()   

  1. Key Laboratory of Industrial Fermentation Microbiology,Ministry of Education,Tianjin Key Laboratory of Industrial Microbiology,College of Biotechnology,Tianjin University of Science & Technology,Tianjin 300457
  • Received:2022-03-06 Online:2022-11-26 Published:2022-12-01
  • Contact: LIU Hao E-mail:guoyufeis@163.com;liuhao@tust.edu.cn

Abstract:

D-glucaric acid, a dicarboxylic acid derivative of glucose, is an important platform compound and is used in medicine, chemical industry and other fields. Using Aspergillus niger as the chassis cell for D-glucaric acid production, expressing the uronate dehydrogenase gene ppudh from Pseudomonas putida KT2440 in A. niger, D-glucaric acid biosynthesis with a titer of 18.74 mg/L was successfully achieved. Strengthening D-glucaric acid biosynthesis secretion, by overexpressing the endogenous inositol oxygenase(anmioxA)and inositol-1-phosphate synthase(aninoA)as well as the carboxylate transporter(scJEN1)from Saccharomyces cerevisiae S288C, D-glucaric acid production was improved significantly with a titer of 102.10 mg/L. Further establishing a NAD+ cofactor recycling system by expressing a NADH oxidase(llnox)from Lactococcus lactis subsp. cremoris MG1363 D-glucaric acid yield was improved to 115.65 mg/L. Finally reducing the carbon flux to glycolysis and pentose phosphate pathway enabled the highest D-glucaric acid production of 313.65 mg/L, which lays the foundation for efficient production of glucaric acid and the corresponding downstream chemicals by microorganisms.

Key words: glucaric acid, Aspergillus niger, cofactor recycling system, RNA interference