C4-dicarboxylic acid production by overexpressing the reductive TCA pathway | Zendy

Ting Zhang | Zendy; Chengyu Ge | Zendy; Li Deng | Zendy; Tianwei Tan | Zendy; Fang Wang | Zendy

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C4-dicarboxylic acid production by overexpressing the reductive TCA pathway

Author(s) -

Ting Zhang,

Chengyu Ge,

Li Deng,

Tianwei Tan,

Fang Wang

Publication year - 2015

Publication title -

fems microbiology letters

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 0.899

H-Index - 151

eISSN - 1574-6968

pISSN - 0378-1097

DOI - 10.1093/femsle/fnv052

Subject(s) - dicarboxylic acid , pichia pastoris , succinic acid , fumarase , fumaric acid , pyruvate carboxylase , malic acid , biochemistry , citric acid cycle , chemistry , tricarboxylic acid , pyruvic acid , gene , biology , citric acid , enzyme , recombinant dna

As C4-dicarboxylic acids could replace C4-petrochemicals, the reductive tricarboxylic acid (TCA) pathway was overexpressed in Pichia pastoris for production of the C4-dicarboxylic acids. Three expression cassettes which carried the pyruvate carboxylase gene (pc), the cytoplasmic malate dehydrogenase gene (mdh1) and the retargeted fumarase gene (Tfum) were integrated into the chromosomal DNA of P. pastoris GS115 alone or jointly. Multicopy integrations were screened using quantitative PCR for C4-dicarboxylic acid overaccumulation. The results showed that the highest titer in 96 h of fumaric, malic and succinic acid (0.76, 42.28 and 9.42 g l(-1)) was obtained by co-expression of pc and mdh1 in P. pastoris. This is the first report about multiple genes engineered in P. pastoris for C4-dicarboxylic acid production. The strain Pp-PC-MDH1, moreover, has a significant potential to produce malic acid in aerobic conditions.

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