Detected the protein content and amino acid content of walnut kernels collected from 17 producing areas in Yunnan, and the amino acid model and nutritional value were comprehensively investigated by amino acid classification, amino acid ratio coefficient method and cluster analysis method. The conclusion indicated that the walnuts derived from each of the various regions were all rich in 17 amino acids (tryptophan was not detected), and the total amount of amino acids was 3.35% to 5.14%, the essential amino acid content was 1.13% to 1.76%, the medicinal amino acid content was 67.47% to 68.63% of the total amino acids, the taste amino acid content was 36.86% to 40.63% of the total amino acid content. The essential amino acids in the amino acid model of the tested walnut kernels were not balanced, and the first limiting amino acid was lysine. The Ratio Coefficient Score (SRC) of walnut kernel was 61.48 to 69.54, and its nutritional value was relatively better. According to the cluster analysis results, the quality of the walnut kernels originating from the Niangqing（Yangbi）, Ludian Hemp 1 (Zhaotong), Ningxiang (Changning), Xunuo 1 (Xundian) and Qingfeng 3 (Zhaoren 5) origins were the best among the 17 walnut kernels in Yunnan.