Influence of the expression of cis,cis-muconate isomerase mutant on aromatic compounds metabolism

2021 Fiscal Year Final Research Report

• Project/Area Number: 17K07706
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Applied microbiology
• Research Institution: Hirosaki University
• Principal Investigator: SONOKI TOMONORI 弘前大学, 農学生命科学部, 准教授 (20502264)
• Project Period (FY): 2017-04-01 – 2022-03-31
• Keywords: cis,cis-muconic acid / lignin / Pseudomonas putida / DMAPP phosphorylase / Nudix hydrolase

Outline of Final Research Achievements

Aiming molecular breeding of a muconate-producing bacterial strain, RNA-seq analyses of the strain expressing a mutated-ccMA cycloisoomerase that accumulates muconate from benzoate in a temperature-dependent manner were conducted. It was expected that PP_0036 and PP_4349, which are considered to be essential genes, are involved in the phenotype of temperature-dependent growth on benzoate. In addition, a gene, nudH, whose deduced amino acid sequence shows high identity with that of RNA pyrophosphohydrolase from Escherichia coli, was identified as a dimethylallylpyrophophate phosphorylase. The expression of nudH improved ccMA productivity of the engineered Pseudomonas putida KT2440-based strain. It is suggested that the supply of dimethylallylmonophosphate, a substrate for synthesizing prenylated flavin mononucleotide that is a critical cofactor for protocatechate decarboxylation, a rate-limiting step in ccMA production was increased by means of the expression of nudH.

Free Research Field

応用微生物学

Academic Significance and Societal Importance of the Research Achievements

ムコン酸は，アジピン酸やテレフタル酸など化石資源から合成され広く使用されているポリマー原料へと展開できる基幹化合物である。低炭素社会の実現に向けて，非可食バイオマス由来を原料としたポリマー原料生産の重要性は今後より一層高まると予想される。本研究の成果は，非可食バイオマスからムコン酸を生産する効率を高めるものであり，持続可能なバイオポリマー原料生産技術の創出に貢献する。