Abstract
A model system was employed to study the operating conditions and primary parameters of enzymic hydrolysis of cod proteins. Pancreatin, papain, and bromelain were used to hydrolyse minced cod fillets under controlled conditions and with the rate of hydrolysis being continually monitored via both the pH-stat and TNBS method. The two methods were compared and evaluated. The rate of protein solubilisation was plotted against the degree of hydrolysis (DH). Dry fish protein hydrolysate (FPH) powders having short, medium and high degrees of hydrolysis (DH of approximately 8%, 11% and 16% respectively) were produced and analysed for their molecular weight distribution, using size exclusion chromatography. Almost complete protein solubilisation (75 g soluble protein per kg hydrolysis solution) could be achieved within an hour, at 40oC, at 1% enzyme/substrate ratio (w/w) with papain and bromelain. The pH-stat was found capable of continuously following the rate of hydrolysis but only at low DH. The TNBS could be accurately used even at high DH to estimate the percentage of the peptide bonds cleaved, but required chemical analysis of withdrawn samples.
Highlights
Applications and uses of proteolytic enzymes for the hydrolysis of fish substrates are vast and varied in purpose
The enzymatic hydrolysis should be regarded as a combination of parallel and consecutive occurring reactions [42]
For a given substrate and enzyme at a given pH, the comparison of different hydrolysates was based on the degree of hydrolysis (DH) as it is the primary determinant of the properties of the hydrolysates
Summary
Applications and uses of proteolytic enzymes for the hydrolysis of fish substrates are vast and varied in purpose. There is considerable interest in the use of enzymes as an alternative to present mechanical methods for recovering high quality protein from fish processing waste, or under-utilised fish species [1,2] Intended uses of such hydrolysates range from nutritional and functional additives with emulsification, aeration, bio-active and anti-oxidative properties [3,4,5,6,7,8,9,10,11], to animal feeds [12,13,14,15,16] and microbial growth media [17,18,19,20]. Most commonly these were: papain [15,21,22,23,24], bromelain [21,24,25], ficin [21,23,26], pepsin [13,21,27], trypsin [21,28,29] and pancreatin [30,31,32]
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