ABSTRACT
Aureobasidium pullulans is an environmental fungus known to produce the exopolysaccharide pullulan, which is used in bone repair, drug delivery, hydrogels, nanoparticles, and food applications. The ability to upscale pullulan production is of great interest to many. In this series of experiments, random amplified polymorphic DNA (RAPD) PCR performed on genomic DNA from A. pullulans ATCC 42023 (AP1) showed an 1800 bp DNA band, seen upon gel electrophoresis. This DNA band was isolated and transformed into either a pullulan-deficient mutant A. pullulans strain or the Saccharomyces cerevisiae expression vector pYES2 using either Novozyme or electroporation, and then pullulan elaboration was determined. The goal of identifying a synthetase gene responsible for pullulan elaboration appears to have been accomplished by utilizing a 1500 bp band obtained by using an RAPD primer in PCR and transforming it into an expression vector via electroporation into S. cerevisiae.
