Background: Only limited information is available on the nutrition knowledge of the general Austrian population and how this relates to the nutrition knowledge of health professionals (medical doctors, pharmacists, nutritionist, dieticians) and school teachers. Methods: Adolescents and adults at the age of 14–75 years (n = 1000), medical doctors (n = 307), pharmacists (n = 295), nutritionists (n = 124), dieticians (n = 160) and school teachers (n = 873) completed an online survey using a German version of the General Nutrition Knowledge Questionnaire-Revised (GNKQ-R) including self-reported data on sex, age, BMI, and health status. Adolescent and adult participants were recruited by a research agency to be representative for the Austrian population for age, sex, and education. A convenience sample was used for health professionals (medical doctors, pharmacists, nutritionist, dieticians) and school teachers. All participants completed a computer-assisted web-based interviewing (CAWI) survey. Results: Total scores for nutrition knowledge of the general population (61.4%) were significantly lower than scores from all other groups (medical doctors 81.3%, pharmacists 83.0%, dieticians 87.5%, nutritionists 85.6%, school teachers 74.5%). The main drivers for better nutritional knowledge were female sex, higher age, and higher level of education, while BMI classification and self-reported health status had no impact. In regard to single questions, the most striking result was the misclassification of sugar as the nutrient with the most calories by 41.4% of the general population while only 29.0% correctly identified fat to be the nutrient with the most calories. Conclusions: The nutritional knowledge of the general population should be significantly improved in order to lay a basis for better dietary behavior. In view of the relatively low scores of teachers, their nutrition education should be improved in order to enable transfer of sound education in schools.