Brewer’s spent grain is the major by-product in beer production. It is produced in large quantities (20 kg per 100 liters of produced beer) throughout the year at a low cost or no cost, and due to its high protein and carbohydrates content it can be used as a raw material in biotechnology. Biotechnological processes based on renewable agro-industrial by-products have ecological (zero CO2 emission, eco-friendly by-products) and economical (cheap raw materials and reduction of storage costs) advantages. The use of brewer’s spent grain is still limited, being basically used as animal feed. Researchers are trying to improve the application of brewer’s spent grain by finding alternative uses apart from the current general use as an animal feed. Its possible applications are in human nutrition, as a raw material in biotechnology, energy production, charcoal production, paper manufacture, as a brick component, and adsorbent. In biotechnology brewer’s spent grain could be used as a substrate for cultivation of microorganisms and enzyme production, additive of yeast carrier in beer fermentation, raw material in production of lactic acid, bioethanol, biogas, phenolic acids, xylitol, and pullulan. Some possible applications for brewer’s spent grain are described in this article including pre-treatment conditions (different procedures for polysaccharides, hemicelluloses, and cellulose hydrolysis), working microorganisms, fermentation parameters and obtained yields. The chemical composition of brewer’s spent grain varies according to barley variety, harvesting time, malting and mashing conditions, and a quality and type of unmalted raw material used in beer production. Brewer’s spent grain is lignocellulosic material rich in protein and fibre, which account for approximately 20 and 70% of its composition, respectively.