Brashears et al. (9) suggested that maximum cholesterol was removed after 20 hr of growth for all cultures tested. In the present study, highest cholesterol removal was determined by the B3 strain for each cell type (growing, resting, and heat-killed). Cholesterol removal capacity of the dead and resting cells implied that cholesterol might

be removed via binding to cells. This result also suggests that higher cholesterol removal by the strains was a result of their growth. Depending on these findings, it can be theorized that even non-viable cells of these strains can be used as cholesterol-reducing probiotic cultures in the gastrointestinal system. Llong and Shah (30) suggested that cholesterol ZD1839 assimilation by growing cells BKM120 was significantly higher than in resting and dead counterparts; however, there was no significant difference reported in the level of cholesterol removal by resting and dead cells. There are two possible mechanisms underlying the ability of lactococci to remove cholesterol from media. One is adhesion of the cholesterol to the cell surface, which is a physical phenomenon and is related to the cell wall. The other possible mechanism is an assimilation of cholesterol by the cells (1). In the present study, because even the heat-killed cells of each strain could remove cholesterol from the media, it seemed that some cholesterol had bound to

the cells. A significant correlation was found between EPS production capacity and cholesterol removal rate for each strain. Generally, strains producing a high amount of EPS (B3, G11, and ATCC 11842) removed much more cholesterol from the medium compared to those having

low EPS production capacity (B2 and A13). These results suggest that the EPS produced by the bacteria interacted with the cholesterol in the medium and bound it in a manner like a dietary fiber. A study by Nakajima et al. (8) revealed that the consumption of milk fermented with an EPS-producing bacterium significantly decreased serum cholesterol levels in rats, whereas Dichloromethane dehalogenase the consumption of milk fermented with a non-EPS-producing strain did not. The researchers reported that slime materials produced by the test bacteria had a beneficial effect on rat cholesterol metabolism. In another study, it was suggested that cholesterol incorporated into, or adhered to, bacterial cells would likely be less available for absorption from the intestines into the blood (9). In our study, most of the cholesterol removed by the strains was recovered with the resuspended cells. Thus, it was not entirely metabolically degraded. However, it is likely that a small portion of the cholesterol that was not recovered from the cell pellets or spent broth was metabolically degraded. These results indicate that the cholesterol in the medium is expected to adhere to the EPS bound to the cell wall. Cholesterol had a positive effect on EPS production in this study.