Outer membrane porin (OMP) expression has been shown to play an important role in antimicrobial resistance. In this study, we observed that OmpK35 of Klebsiella pneumoniae had varied expression profiles in different nutrient broths. the potential factors that could influence protein expression were assessed. K. pneumoniae (ATCC 13883) was cultured into two commercial available nutrient broths and also into solutions of the individual ingredients. To ensure that OmpK35 was detected, an OmpK35 deficient mutant was generated as control. When OmpK protein expression profiles were analyzed by SDS-PAGE, OmpK35 exhibited two different isoforms. Expression of an additional isoform-like OmpK35 protein was identified in one of the broths. No OmpK35 isoforms were observed when the individual ingredients of beef extract, casein or gelatin were used as culture medium. OmpK35 isoform expression could be repressed by adding more beef extract. In summary, OmpK can exhibit varied protein expression profiles when growing in different nutrient broths. The isoform-like protein expression of OmpK35 may lead to confusion in OmpK protein analysis.