| Products/Services Used | Details | Operation |
|---|---|---|
| Gene Synthesis | The pJFE12 vector was constructed by replacing the ADH1p-Gal4-ADH1t cassette with TEF1p-CYC1t into SphI-digested plasmid pGBT9 (Fields and Song, 1989). Genes AroZ-Neu, AroY-Com, and CatA-Cup were codon-optimized for their expression in S. cerevisiae and synthesized from GenScript (Singapore). TDH3p, PGK1p and CYC1t were amplified from the genomic DNA of S. cerevisiae. Gene fragments, AroZ-CYC1t, TDH3p-AroY-CYC1t and PGK1p-CatA, were fused through OE-PCR and sequentially cloned into plasmid pJFE12, resulting in plasmid pJAA9N. | Get A Quote |
New exogenous cis,cis-muconic acid biosynthetic pathway genes were expressed in Saccharomyces cerevisiae. The xylose isomerase gene from Bacteroides valgutus and pentose phosphate pathway genes from S. cerevisiae were overexpressed in the yeast strain. The strain was further modified by the overexpression of gene Aro1 (with a stop codon of AroE) and a feedback-resistant Aro4opt mutant gene from S. cerevisiae. Under oxygen-limited conditions, it produced 65 mg/L cis,cis-muconic acid from xylose. Co-fermentation of 88 g/L glucose and 50 g/L xylose generated 54 g/L ethanol and 248 mg/L cis,cis-muconic acid. Under aerobic conditions, muconic acid titer reached 424 mg/L. With the supplement of 1 g/L catechol, 1286 m... More
