Collagenolytic Activity of Some Marine Bacteria

Abstract

Reconstituted, acid-extracted collagen was used to prepare a medium to screen proteolytic marine bacteria for their ability to elaborate collagenolytic enzymes. The medium was resistant to solubilization by trypsin, hyaluronidase, chondroitinase ABC, and various marine proteinases, but was readily hydrolyzed by commercial Clostridium collagenases. Eighty-seven marine isolates collected in the vicinity of Bermuda, Oahu (Hawaii), and Stone Harbor and Cape May, N.J., were screened. Approximately 44% of the isolates were capable of elaborating enzymes that hydrolyzed reconstituted collagen gels. Several cultures produced collagenolytic enzymes only when grown in the presence of collagen or degradation products of collagen, and with very few exceptions the presence of collagen in the medium greatly enhanced collagenolytic enzyme production. The enzymes from a collagenolytic Bermuda marine isolate were studied in more detail to illustrate that the enzymes capable of hydrolyzing reconstituted collagen were separable from nonspecific proteinases by zone electrophoresis and that these enzymes were true collagenases by virtue of their ability to hydrolyze native bovine Achilles' tendon obtained from three different sources.

Purification and characterization of a marine bacterial collagenase

“…Enzyme Production and Purification. The bacterium grows well and produces proteolytic enzymes on a variety of hydrolyzed casein media, but collagenase production occurs only when induced by the addition of collagen or certain collagenderived products to the media (Merkel et al, 1975). Maximum collagenase production in collagen or gelatin-containing media occurred in 23-24 h of incubation at 25 °C in shaker flasks and in 10-12 h of incubation in a New Brunswick Scientific 7.5-L fermentor.…”

“…After incubation for about 2 h at 37 °C, two distinct zones of digestion appeared which coincided with the two stained bands. When similar gel columns were placed on an algal chromoprotein plate (Merkel, 1966), these two bands had no activity against the chromoproteins which ruled out their being non-specific proteinases. Purified Vibrio B-30 collagenase has no or negligible hydrolytic activity against algal chromoproteins, urea-denatured hemoglobin, or casein.…”

“…Acid-extracted, calfskin collagen was prepared from freshly slaughtered calves by a modification (Merkel et al, 1975) of the procedure of Gross & Kirk (1958). Laboratory scale enzyme production was accomplished in 7.5-L fermentors (New Brunswick Sc.…”

“…(1) Reconstituted collagen plate method (Merkel et al, 1975) was used as a semiquantitative measure of collagenase activity in various enzyme preparations.…”

Bioplastics in the Sea: Rapid In-Vitro Evaluation of Degradability and Persistence at Natural Temperatures

“…Collagens are hydrolyzed by collagenolytic proteases (Ran et al, 2013). Only a few studies addressed collagenolytic enzymes in marine bacteria (Merkel et al, 1975;Kurata et al, 2007;Zhao et al, 2008). Our experiment showed a substantial degradation of collagen by the protease but not by the esterase and the lipase.…”