Diabetes mellitus is a risk factor for development of periodontal disease with progressive periodontal destruction seen in diabetic patients. Advances in oral and periodontal disease diagnostic research are moving towards methods whereby periodontal risk can be identified and quantified by objective measures such as biomarkers. The aim of this study was to identify the presence of beta glucuronidase activity as an enzymatic biomarker of periodontal tissue destruction in patients of Diabetes mellitus associated with Periodontitis. β Glucuronidase activitywas estimated in saliva, GCF (Gingival Crevicular Fluid) and serum in healthy subjects and patients with Diabetes mellitus, Periodontitis and Diabetes associated periodontitis. Enzyme activity was compared with clinical parameters like Probing pocket depth, Plaque Index and Gingival Index. It was maximally raised in diabetic cases with periodontitis. Enzyme activity was found to be highest in crevicular fluid compared to saliva and serum. However, serum Glucuronidase correlated significantly with clinical indices. β-glucuronidase may be employed on routine basis as a chair side test for screening and diagnosis of patients with periodontitis in diabetics.