Guava (Psidium guajava) contain a great deal of polyphenol compound and work on the treatment of Diabetes mellitus effectively. In this study, the bioactivities of aqueous extract (GLEx) of guava leaf were investigated. Total phenolic contents of GLEx was 266.9 mg tan/g. The effects of GLEx on digestive enzymes, α-amylase, maltase and sucrase were investigated. IC50 values of GLEx against α-amylase, maltase and sucrase were 0.65 mg/ml, 2.0 mg/ml and 3.5 mg/ml respectively. The effect of GLEx on oral glucose tolerance test (OGTT) was performed in normal ICR mouse, control (dstilled water) and GLEx (aqueous extract of Guava leaf) were co-administered orally with glucose, showed reducing effect on the blood glucose level. The guava is likely to useful for prevention or improvement of hyperglycemia by lowering the blood glucose level and inhibiting glycoside hydrolase activity.