The invention discloses a fluorescence analysis method of pichia pastoris recombinant protein secretion expression quantity, and relates to the technical field of gene engineering. The pichia pastorisendoplasmic reticulum membrane protein is used with expression fluorescent protein, and the secretion expression quantity of the recombinant protein is represented by detecting the fluorescence valueof the thallus. Fluorescent protein (EGFP) fluorescent value can be used for rapidly analyzing the secretion expression quantity of the recombinant protein in pichia pastoris, and a complicated operation is not needed, so that the method is simple, convenient and rapid, and is suitable for detecting the secretion expression quantity of the recombinant protein in bioengineering and genetic engineering, and the rapid detection of the recombinant protein recombinant strain of the efficient secretion and expression recombinant protein in pichia pastoris is facilitated.