The Optimization Technology Of High F Value Oligopeptides Produced From Swine Blood

In order to make full use of livestock blood proteins and develop functional bloodingbiological materials, researches on the technology optimization for preparing plasmaprotein by enzyme and decolouring hydrolyzate by active carbon from swine plasma werecarried out.An enzymolysis approach based on the quaternionic linear regression orthogonaldesign with four variables of enzyme dose, enzymolysis temperature, enzymolysis timeand pH on the degree of hydrolysis with Alcasale was employed to optimize preparationparameters of plasma protein. And a decoloration approach based on the quaternioniclinear regression orthogonal design with three factors of decoloration temperature, pH andtime was employed to optimize the decolorization parameters.The results were as follows:the optimal enzymolysis technology was that the swine plasma was hydrolyzed by6%enzyme dose at60℃and pH9.5for3.67h, under these conditions,30.99%swine plasmawere hydrolyzed. The optimal decoloration technological parameters were active carbon2%, decoloration temperature48℃, pH2.8, decolorationduration29min, which resulteda high decolouring rate of76.06%and a high protein retention rate of82.24%.Enzyme immobilization used chitosan as the carrier, glutaraldehyde as thecrosslinking agent. The immobilization conditions such as temperature, pH andimmobilied time were optimized by orthogonal design. It is shown from range analysisthat the optimum process were temperature40℃,immobilization time9h and pH9. Thefree loss of the immobilized enzyme during the144h were2～3%, the immobilizedenzyme was stable and could be used widely. Five replicated experiments were carried out to calculate the relative differencesbetween the test trial and the control trial. The relative differences of Hemoglobinoligopeptides was1.07%, and the relative differences of blood-red oligopeptides0.17%.The absorbance at220nm was used as the determination index. The peptides fromporcine plasma protein and hemoglobin were purified by a chromatography a columnfilled with G-15gel which was eluted by distilled water. The molecular weight wasdetermined by using high performance liquid chromatography. The molecular weight ofthe high Fisher ratio oligopeptide derived from porcine plasma protein ranged from610.5to1214.1Da. And the range of molecular weight of high Fisher ratio oligopeptide fromprocine hemoglobin was635.7～1151.1Da.The amino acid composition of oligopeptide derived from porcine plasma andhemoglobin was determined on a Automatic Amino Acid Analyzer. Then the Fisher valuesof the two high Fisher ratio oligopeptides were calculated to be30.51and38.56, whichwere increased by3.59times and7.38, respectively. The results of purification weresignificant and Fisher-values were in accordance with the requirement that the high Fisherradio bioactive peptide should be higher than or at least equal to20.The acute toxicity, hereditary toxicity and30-day feed assay were employed toevaluate the safety of high Fisher radio oligopeptides from porcine plasma protein andhemoglobin with the purpose of providing evidence for the development and safe intake offunctional food made of active peptides from animal blood.