Engineering Saccharomyces cerevisiae to produce feruloyl esterase for the release of ferulic acid from switchgrass
| Autor: | Sarah B. Batt, Gautam Sarath, Hans Liao, Dominic W. S. Wong, Victor J. Chan |
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| Rok vydání: | 2010 |
| Předmět: |
chemistry.chemical_classification
Expression vector biology Coumaric Acids Aspergillus niger Saccharomyces cerevisiae Substrate (chemistry) Bioengineering biology.organism_classification Nitro Compounds Panicum Applied Microbiology and Biotechnology Yeast Catalysis Ferulic acid chemistry.chemical_compound Enzyme Biochemistry chemistry Feruloyl esterase Carboxylic Ester Hydrolases Biotechnology |
| Zdroj: | Journal of industrial microbiologybiotechnology. 38(12) |
| ISSN: | 1476-5535 |
| Popis: | The Aspergillus niger feruloyl esterase gene (faeA) was cloned into Saccharomyces cerevisiae via a yeast expression vector, resulting in efficient expression and secretion of the enzyme in the medium with a yield of ~2 mg/l. The recombinant enzyme was purified to homogeneity by anion-exchange and hydrophobic interaction chromatography. The specific activity was determined to be 8,200 U/μg (pH 6.5, 20°C, 3.5 mM 4-nitrophenyl ferulate). The protein had a correct N-terminal sequence of ASTQGISEDLY, indicating that the signal peptide was properly processed. The FAE exhibited an optimum pH of 6–7 and operated optimally at 50°C using ground switchgrass as the substrate. The yeast clone was demonstrated to catalyze the release of ferulic acid continuously from switchgrass in YNB medium at 30°C. This work represents the first report on engineering yeast for the breakdown of ferulic acid crosslink to facilitate consolidated bioprocessing. |
| Databáze: | OpenAIRE |
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