The article presents an overview of the latest advances in investigations of the biosynthesis, molecular properties, and associated biological activity of pullulan. The literature survey on the pullulan biosynthesis is intended to illustrate how the great variety of environmental conditions as well as variability in strain characteristics influences the metabolic pathways of the pullulan formation and effects structural composition of the biopolymer. Molecular properties of pullulan as alpha-(1-->4)- and alpha-(1-->6)-glucan are discussed in terms of similarities with amylose and dextran structures, and an emphasis is made on the inherent biological activity of pullulan molecules. The author also attempts to summarize the concepts, options, and strategies in chemical modification of the biopolymer and to delineate future prospects in designing new biologically active derivatives.