Dietary proteins influence colorectal cancer (CRC) risk, depending on their quantity and quality. Here, using pyrosequencing, we compared the fecal microbiota composition in Balb/c mice fed either a normal protein/carbohydrate diet (ND, 20% casein and 68% carbohydrate) or a high-protein/low-carbohydrate diet (HPLCD, 30% casein and 57% carbohydrate). The results showed that HPLCD feeding for 2 weeks reduced the diversity and altered the composition of the microbiota compared with the ND mice, which included a decrease in the proportion of the family Lachnospiraceae and Ruminococcaceae and increases in the proportions of the genus Bacteroides and Parabacteroides, especially the species EF09600_s and EF604598_s. Similar changes were reported in patients with inflammatory bowel disease, and in mouse models of CRC and colitis, respectively. This suggests that HPLCD may lead to a deleterious luminal environment and may have adverse effects on the intestinal health of individuals consuming such a diet.

Bacillus subtilis HK176 with high fibrinolytic activity was isolated from cheonggukjang, a Korean fermented soyfood. A gene, aprE176, encoding the major fibrinolytic enzyme was cloned from B. subtilis HK176 and overexpressed in E. coli BL21(DE3) using plasmid pET26b(+). The specific activity of purified AprE176 was 216.8 ± 5.4 plasmin unit/mg protein and the optimum pH and temperature were pH 8.0 and 40°C, respectively. Error-prone PCR was performed for aprE176, and the PCR products were introduced into E. coli BL21(DE3) after ligation with pET26b(+). Mutants showing enhanced fibrinolytic activities were screened first using skim-milk plates and then fibrin plates. Among the mutants, M179 showed the highest activity on a fibrin plate and it had one amino acid substitution (A176T). The specific activity of M179 was 2.2-fold higher than that of the wild-type enzyme, but the catalytic efficiency (kcat/Km) of M179 was not different from the wild-type enzyme owing to reduced substrate affinity. Interestingly, M179 showed increased thermostability. M179 retained 36% of activity after 5 h at 45°C, whereas AprE176 retained only 11%. Molecular modeling analysis suggested that the 176(th) residue of M179, threonine, was located near the cation-binding site compared with the wild type. This probably caused tight binding of M179 with Ca(2+), which increased the thermostability of M179.

Jihyun Ahn and Jae-Yong Park. Prev Nutr Food Sci 2023;28:492-501. https://doi.org/10.3746/pnf.2023.28.4.492