Exo-maltotetraohydrolase produced by Pseudomonas stutzeri IAM 12097 was characterized with respect to substrate specificity, the reaction products and hydolysis rate on various carbohydrates. Maltopentaose, maltohegtaose, soluble starch, amylose, amylopectin, oyster glycogen and gelatinized starch of corn, potato, glutinous rice, green banana and arrow root were hydolyzed by this enzyme, but α, β, γ-cyclodextin, sucrose, raffinose, lactose pullulan, maltose, maltotriose and maltotetraose were not hydolyzed. Among oligosaccharides, maltohexaose was favorably hydrolyzed by this enzyme and the main reaction product of oligosaccharides and polysaccharides was maltotetraose. Addition of pullulanase to this enzyme increased the hydolysis rate on gelatinized starches, but it did not on raw starches. Among various starches, corn starch was favorably hydrolyzed by this enzyme, whereas it acted on potato starch, arrow root starch and high amylose corn starch weakly.