As a specific spoilage organism of refrigerated fish， Shewanella putrefaciens can survive longer during cold storage due to its strong adaptability to cold and cause the fish spoilage. Cold shock protein （CSP） is one of the key proteins that affects the adaptability of bacteria to low temperature. In the present study， three CSP genes were obtained by PCR amplification， and their structure and function were analyzed by bioinformatics. The results showed that the Csp1589 and Csp1376 of S. putrefaciens shared close homology with Escherichia coli CspA， while Csp2252 was more homology with E. coli CspD； the CSPs were small hydrophilic acidic protein with molecular weight of about 7.4 ku， all of which were located in the cytoplasm and are non-secretory and non-transmembrane proteins. All of them possess specific cold shock domain， and sites and motifs that could bind to single-stranded DNA and RNA. The secondary and tertiary structures of CSPs conformed to the characteristics of CSP family， which contained five reverse parallel β-strands. Three CSPs were functionally predicted to interact with various proteins， including histidine kinase involved in two-component signal transduction system， DNA polymerase involved in DNA replication， chaperone protein htpG， etc. And the possible ligand binding sites of CSP were predicted. It provided some references for further elucidation of the function of CSPs in the process of low temperature adaptation of S. putrefaciens.