Chitin-glucan complex (CGC) copolymer is a cell wall component in fungi and yeasts, used in medicine and cosmetics applications. This study investigated the production of CGC from Aureobasidium pullulans KSY-0516, which is used in the industrial production of β-1,3/1,6-glucan. The amino acid composition of the A. pullulans fermentation by-products was determined using an amino acid analyzer. The total amount of amino acid was 6.67 g/100 g of by-product. Additionally, the recovery of the β-glucan complex from the A. pullulans fermentation by-products was 8.44 g/100 g of by-product using ethanol precipitation. The CGC yield was determined from the by-products after yeast fermentation. The recovery of CGC was 14.83% for the yeast cell wall obtained by alkaline extraction. The CGC and β?glucan were characterized using Fourier transform infrared analysis, and compared with crab shell chitin and laminarin. This study aimed to examine the potential, as a useful substrate for biotechnology, by producing CGC from yeast cultivation by-products.