The invention relates to beaded electrospun nanofiber immobilized saccharomycete and a preparation method thereof. Nanofiber comprises electrospun polymer or macromolecules, a solvent and a saccharomycete; prepared mixed solution is placed in an injector and is spun with an electrospinning instrument, microorganisms are introduced into the nanofiber by dispersion of the saccharomycete in a spinning solution, the advantages of the nanofiber such as high specific area, high porosity, large length-diameter ratio and bulkiness and porousness are combined, in-situ immobilization is achieved for saccharomycete cells, an active organic/microorganism composite is obtained, and a function similar to a multi-enzyme bioreactor is enabled. A product and preparation method of the invention have the advantages such as high processing efficiency, high reacting speed, ease of control, high antifouling capacity, continuous reusability, and ease of product separation. The beaded electrospun nanofiber immobilized saccharomycete is applicable to the fields such as biological catalysis, filtering and adsorption, drug delivery and tissue engineering.