Non-Saccharomyces yeasts are of growing interest as probiotic candidates, but strain-level charac-terization remains limited. We characterized Pichia kluyveri PK88™, isolated from coffee beans (NRRL deposit # 68561), using phenotypic assays, whole-genome sequencing, and in silico screen-ing. PK88™ was non-hemolytic and produced no detectable patulin, ochratoxin A, or aflatoxins. Genome analysis confirmed species identity (98.5–98.8% average nucleotide identity) and identi-fied no acquired antimicrobial-resistance genes, canonical virulence factors, or decarboxylases for harmful biogenic amines. By high-inoculum broth microdilution, MICs were low for echi-nocandins and amphotericin B and higher for fluconazole and posaconazole. Measured survival after acid, bile and anaerobic exposure was retained (93% at pH 2; 82–84% in bile), although sur-vival at pH 1 and under anaerobiosis was lower; hydrophobicity fell from 91% to 73–77% and auto-aggregation rose to 98–100%. PK88 supernatant inhibited Escherichia coli and Candida albi-cans in preliminary assays. In two independent experiments, Caco-2 cells exposed to PK88™ showed cytotoxicity ≤20% in every replicate up to 6 × 104 organisms/well and variable increases at higher doses. These preliminary data support further evaluation.