Aspevik Tone, Egede-Nissen Henning, Oterhals Ĺge
Nofima, Kjerreidviken 16, N-5141 Fyllingsdalen, Norway.
Food Technol Biotechnol. 2016 Dec;54(4):421-431. doi: 10.17113/ftb.54.04.16.4553.
The hydrolytic and cost efficiencies of five endopeptidases (Alcalase 2.4L, Corolase 7089, Neutrase 0.8L, Promod 671L and Protex 7L) to hydrolyze Atlantic salmon by-products were compared at standardized activity levels based on a casein assay. The substrate was characterized prior to the hydrolytic experiments (pH=6.5, =50 °C) to obtain substrate--specific constants for nitrogen to protein mass (in g) ratio, conversion factor =5.23 and total amount of peptide bonds ()=9.3 mmol per g of protein. At low enzyme activity to substrate ratio, all enzymes were equally efficient in hydrolyzing the substrate. At highest enzyme activity to substrate ratio, Protex 7L, Alcalase 2.4L and Promod 671L gave higher degree of hydrolysis (DH=14.2-14.6%) than Corolase 7089 (13.2%) and Neutrase 0.8L (11.6%) after 120 min of hydrolysis. No differences were observed in protein recovery (yield of solubilized protein) relative to DH. Determination of DH was followed by the pH-STAT and -phthaldialdehyde methods. Based on pH-STAT data, response surface regression models were established based on the combined effects of hydrolysis time and enzyme activity to substrate ratio on DH and protein recovery. The modelling approach was combined with enzyme cost to identify the most cost-efficient enzyme (Protex 7L).
基于酪蛋白测定法,在标准化活性水平下比较了5种内肽酶(碱性蛋白酶2.4L、科罗蛋白酶7089、中性蛋白酶0.8L、普洛酶671L和普罗泰克斯7L)对大西洋鲑鱼副产物的水解效率和成本效率。在水解实验(pH = 6.5,温度 = 50°C)之前对底物进行了表征,以获得底物特定的常数,即氮与蛋白质质量(以克为单位)的比率、换算系数 = 5.23以及每克蛋白质的肽键总量()= 9.3 mmol。在低酶活性与底物比率下,所有酶在水解底物方面效率相同。在最高酶活性与底物比率下,水解120分钟后,普罗泰克斯7L、碱性蛋白酶2.4L和普洛酶671L的水解度(DH = 14.2 - 14.6%)高于科罗蛋白酶7089(13.2%)和中性蛋白酶0.8L(11.6%)。相对于水解度,在蛋白质回收率(溶解蛋白质的产率)方面未观察到差异。采用pH计法和邻苯二甲醛法测定水解度。基于pH计法数据,根据水解时间和酶活性与底物比率对水解度和蛋白质回收率的综合影响建立了响应面回归模型。该建模方法与酶成本相结合,以确定最具成本效益的酶(普罗泰克斯7L)。