Influence of mucin on glycosidase, protease and arylamidase activities of human gut bacteria grown in a 3-stage continuous culture system.
Human intestinal bacteria were grown in a 3-stage continuous culture system on a medium containing complex polysaccharides and proteins as carbon and nitrogen sources. Selected bacterial populations were enumerated and glycosidase, protease and arylamidase activities measured. Comparison of arylamidase and glycosidase activities in the multichamber system (MCS) and faeces showed that the predominant faecal enzymes were also produced by bacteria growing in the MCS. After 48 d operation, porcine gastric mucin (5.8 g/d) was independently fed to vessel 1. Elevated levels of volatile fatty acid (VFA) formation showed that the glycoprotein was actively fermented. The increase in carbohydrate availability as a result of breakdown of the mucin oligosaccharides stimulated bacterial growth and activities. The enzymological measurements showed that mucin increased production of both cell-bound and extracellular glycosidases, such as beta-galactosidase, alpha-glucosidase and N-acetyl-beta-glucosaminidase. Protease activities were profoundly influenced by mucin. These were largely cell-bound in non-mucin cultures but were predominantly extracellular and collagenolytic when mucin was present. Experiments with protease inhibitors showed that cysteine proteases were the major cell-bound and extracellular enzymes in both mucin and non-mucin cultures, but that serine and metalloproteases were also present. The effect of mucin on arylamidase formation was less marked, although there was increased production of these enzymes in vessels 1 and 2 of the MCS. These results suggest that host-produced substances such as mucin glycoprotein may play a role in modulating the growth and activity of bacteria growing in the human large intestine.
Macfarlane
G T
GT
Medical Research Council, Dunn Clinical Nutrition Centre, Cambridge, UK.
Hay
S
S
Gibson
G R
GR
D000626:Aminopeptidases / Q000378:metabolism*; D001421:Bacteria, Anaerobic / Q000201:enzymology* / Q000254:growth & development; D003470:Culture Media; D005232:Fatty Acids, Volatile / Q000096:biosynthesis; D005285:Fermentation; D006026:Glycoside Hydrolases / Q000378:metabolism*; D006801:Humans; D007420:Intestine, Large / Q000382:microbiology; D009077:Mucins / Q000378:metabolism*; D010447:Peptide Hydrolases / Q000378:metabolism*
D003470:Culture Media; D005232:Fatty Acids, Volatile; D009077:Mucins; D006026:Glycoside Hydrolases; D010447:Peptide Hydrolases; D000626:Aminopeptidases