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In times where overweight and diabetes are major health issues, the demand for taste–optimized low-calorie sweeteners and sweetness enhancers is increasing. The consumer’s preference for natural food ingredients has enforced the search for natural sweeteners. A potential source of such a natural non-nutritive sweetener is the Vietnamese plant Mycetia balansae, which is used for sweetening by locals. We have identified the sweet principle of Mycetia balansae using sensory-guided analysis and characterized its action on the human sweet taste receptor with an integrated approach combining homology modelling and cell-based functional receptor expression.