Partial Characterization of a Crude Enzyme Extract from the Freshwater Prawn, Macrobrachium rosenbergii

Abstract

The activity of a crude enzyme preparation.extracted from hepatopancreas of the freshwater prawn, Macrobrachium-rosenbergii, was assayed for collagenolytic, trypsinolytic, orchymotrypsinolytic, and pepsinolytic activities against collagen, lyophilized prawn tissue, and artificial substrates. At optimum pH for each activity, the enzyme preparation had collagenolytic activitiy, slight trypsinolytic and ol-chymotrypsinolytic activities; and no pepsinolytic activity. Of the commercial enzymes tested, only collagenase significantly degraded lyoph@zed prawn tissue. These results suggest that the -. prawn enzyme preparation may contain a collagenolytic portion which might affect the texture of the prawn.

Extraction and Purification of Collagenase Enzymes: A Critical Review

“…Since tissue collagenases are digestive enzymes, they are commonly isolated from the digestive tracts of various fish and invertebrates including: tadpole tailfin (Gross and Nagai, 1965;Nagai et al, 1966), rabbit skin (Fullmer and Gibson, 1966), rat uterus (Jeffrey and Gross, 1967), rheumatoid synovial tissue (Evanson et al, 1968), mouse bones (Sakamoto et al, 1972), crabs (Eizen and Jeffrey, 1969;Grant et al, 1983;Klimova et al, 1990;Sellos and Van Wormhoudt, 1992;Gerasimova and Kupina, 1996;Zefirova et al, 1996), fresh water prawns (Baranowski et al, 1984), crayfish (Garcia-Carreno et al, 1994), Atlantic cod (Kristjánsson, et al, 1995), tropical shrimp (Penaeus vannamei) Van Wormhoudt et al, 1992) and catfish (Parasilurus asotus) (Klimova et al,1990;Sellos and Van Wormhoudt, 1992).…”

“…Collagenolytic serine proteases (EC 3.4.21.32) were first isolated from the fiddler crab (Uca pugzlator) hepatopancreas (Eisen et al, 1973) but have now been extracted and characterized from the digestive tracts of a variety of fish and aquatic invertebrates (Eizen and Jeffrey, 1969;Grant et al, 1983;Baranowski et al, 1984;Garcia-Carreno et al, 1994;Kristjánsson et al, 1995;Gerasimova and Kupina, 1996;Zefirova et al, 1996). Serine collagenases are much less abundant than metallo collagenases (Gonzales and Robert-Baudouy, 1996).…”

“…5). Several other authors (Baranowski et al, 1984;Teruel and Simpson, 1995;Hernández-Herrero et al, 2003;Burgos-Hernández et al, 2005) employed a very similar technique to extract collagenase from a variety of fish tissues including cod and winter flounder (Pseudopleuronectes americanus) muscles and shrimp hepatopancreas, with the tris-HCl buffer containing CaCl 2 but at a pH of 7.4, fractionation with ammonium sulfate (40-80%) and incorporating a filtration step prior to the initial centrifugation (Fig. 6).…”

“…6: The Tris-buffer extraction and fractionation system of collagenase enzyme from freshwater prawn (Baranowski et al, 1984) and winter flounder fish skeletal muscle (Teruel and Simpson, 1995) °° Collagenase purification: Once a crude collagenase extract is recovered, it must be purified using one of several chromatographic methods that can be classified as: gel filtration, ion exchange, hydrophobic interaction or affinity Figure 9 shows the principle of these purification processes. It should be noted that these chromatography techniques are commonly coupled together so that most purification schemes will begin with gel filtration, followed by either ion exchange or affinity chromatography or both, depending on the application.…”

Quality Changes in Australian Red Claw Crayfish,Cherax quadricarinatus, Stored at 0°C

“…Studies on freshwater prawn, Macrobrachium rosenbergii, have shown that storage of intact freshwater prawn on ice resulted in muscle "mushiness" due to diffusion of proteolytic and collagenolytic enzymes from the hepatopancreas (Baranowski et al 1984;Lindner et al 1988;Nip and Moy 1988). However, such texture deterioration was completely eliminated when prawn "heads" were removed prior to storage (unpublished results).…”