Abstract
Akkermansia muciniphila can produce various mucin-degrading proteins. However, the functional characteristics of these proteins and their role in mucin degradation are unclear. Of the predicted protein-coding genes, Amuc_1434, which encodes for a hypothetical protein, is the focus in this study. A recombinant enzyme Amuc_1434 containing the 6× His-tag produced in Escherichia coli (hereinafter termed Amuc_1434*) was isolated to homogeneity and biochemically characterised. Results showed that the enzyme can hydrolyse hemoglobin with an activity of 17.21 U/μg. The optimal pH and temperature for hemoglobin hydrolysis of Amuc_1434* were found to be around 8.0 and 40 °C, respectively. Amuc_1434* is identified as a member of the aspartic protease family through the action of inhibitor pepstatin A. Amuc_1434* promotes the adhesion of colon cancer cell line LS174T, which can highly express Muc2. Significantly Amuc_1434* can degrade Muc2 of colon cancer cells. Amuc_1434 is mainly located in the colon of BALB/c mice. These results suggest that the presence of Amuc_1434 from Akkermansia muciniphila may be correlated with the restoration of gut barrier function by decreasing mucus layer thickness.
Highlights
Mucins are a family of high molecular weight glycosylated proteins that protects epithelial cells and exists on multiple organ surfaces [1,2,3]
Mucin 2 (Muc2) is known as the intestinal mucin, which is associated with intestinal goblet cells, consisting of a variable number of vertically repeated amino acid sequences (VNTR) [8]
It is involved in the occurrence and development of colorectal cancer (CRC), which ranks the third cause of mortality of cancers in the world [14,15]
Summary
Mucins are a family of high molecular weight glycosylated proteins that protects epithelial cells and exists on multiple organ surfaces [1,2,3]. Mucin 2 (Muc2) belongs to the type of secretory mucinous proteins It is normally expressed only in the colon and rectum but not in normal ovarian epithelial cells, and generally considered as an intestinal-associated antigen [7]. Muc is the main component of the protective layer of colon mucosa [11], which can inhibit intestinal inflammatory response, and prevents the development of intestinal tumors [12,13]. It is involved in the occurrence and development of colorectal cancer (CRC), which ranks the third cause of mortality of cancers in the world [14,15]. Muc degradation may be an effective treatment for some cancers
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