Pullulan is an extracellular polysaccharide produced by Aureobasidium pullulans, which is widely used in food, pharmaceutical, and cosmetics industries. To evaluate the effect of vitamin B5 on the pullulan biosynthesis., the changes of key enzyme activities and the protein composition during fermentation were measured, and the protein composition was analyzed in the early (24 h) and late (84 h) stages of fermentation by label-free quantitative proteomics and liquid chromatography-tandem mass spectrometry (LC-MS/MS). Furthermore, the bioinformatics analysis was conducted to analyze the different proteins. The fermentation medium with 0.2g/L vitamin B5 addition could increase the pullulan production from 87.3 to 102 g/L, and decreased the mass average molar mass from 2.14 ×102 kDa to 1.11×102 kDa. In the experimental group, glucose phosphate mutase, uridine diphosphate glucose pyrophosphorylase, glucosyltransferase, α-amylase, and pullulanase activities increased by 6.9%, 19.8%, 20.9%, 20.8%, and 12.1%, respectively. 387 and 381 differentially expressed proteins were identified at two time points (folds>1.5, P<0.05), and the GO function enrichment and KEGG pathway enrichment showed that they were mainly involved in some important biological processes, such as cellular processes and metabolic processes, which mainly regulated the carbon metabolism, amino acid metabolism, and fatty acid metabolism. These changing pathways eventually lead to the increasing production and mass average molar mass of pullulan.
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