This study investigated the effects of valine on growth, intestinal enzyme activities and microflora in juvenile Jian carp (Cyprinus carpio var. Jian). A total of 1200 fish with an average initial weight of 9.67 ± 0.03 g were fed diets containing 5.3 (unsupplemented control), 8.7, 11.8, 14.9, 18.7 and 20.1 g valine kg−1 diet for 60 days. Results indicated that the specific growth rate, feed efficiency, body protein and lipid content of fish were significantly improved by the dietary valine (P < 0.05). The hepatopancreas weight and activities of trypsin, amylase, lipase, chymotrypsin, glutamate oxaloacetate transaminase (GOT) and glutamate pyruvate transaminase (GPT) took the similar trends. Similarly, the optimum levels of dietary valine induced increases in the intestinal length, weight, folds height and activities of alkaline phosphatase, gamma-glutamyl transpeptidase and creatine kinase. In contrast, the trends of muscle GOT activity and plasma ammonia content were opposite. Intestinal Aeromonas, Escherichia coli, Lactobacillus and Bacillus were changed by dietary valine supplementations. The dietary valine requirement for Jian carp (9.67–76.4 g) based on SGR was 13.7 g valine kg−1 diet (4.0 g valine 100 g−1 CP). Together, these results indicated that valine improved fish growth, digestive and absorptive ability.