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Development of a one-step gene knock-out and knock-in method for metabolic engineering of Aureobasidium pullulans.
- Source :
-
Journal of Biotechnology . Jun2017, Vol. 251, p145-150. 6p. - Publication Year :
- 2017
-
Abstract
- Aureobasidium pullulans is an increasingly attractive host for bio-production of pullulan, heavy oil, polymalic acid, and a large spectrum of extracellular enzymes. To date, genetic manipulation of A. pullulans mainly relies on time-consuming conventional restriction enzyme digestion and ligation methods. In this study, we present a one-step homologous recombination-based method for rapid genetic manipulation in A. pullulans . Overlaps measuring >40 bp length and 10 μg DNA segments for homologous recombination provided maximum benefits to transformation of A. pullulans . This optimized method was successfully applied to PKSIII gene (encodes polyketide synthase) knock-out and gltP gene (encodes glycolipid transfer protein) knock-in. After disruption of PKSIII gene, secretion of melanin decreased slightly. The melanin purified from disruptant showed lower reducing capacity compared with that of the parent strain, leading to a decrease in exopolysaccharide production. Knock-in of gltP gene resulted in at least 4.68-fold increase in heavy oil production depending on the carbon source used, indicating that gltP can regulate heavy oil synthesis in A. pullulans . [ABSTRACT FROM AUTHOR]
Details
- Language :
- English
- ISSN :
- 01681656
- Volume :
- 251
- Database :
- Academic Search Index
- Journal :
- Journal of Biotechnology
- Publication Type :
- Academic Journal
- Accession number :
- 123268855
- Full Text :
- https://doi.org/10.1016/j.jbiotec.2017.04.029