Studies On The Preparation, Structure, And Function Of Hemi Peptide-Fe And Its Application In Fruit And Vegetable Beverages

The hemi iron in porcine blood was able to be utilized broadly in foods such as beverage and soy sauce. In this paper, Hemi Peptide-Fe was prepared from porcine Hb by ways of both LHPEF and Trypsin. This task studied on the extraction of high-purity Hemi Peptide-Fe from enzymolysis solution by MCAC, and it's molecular structure, inoxidability in vitro and MBF flocculation. In order to solve these problems, such as long isolation time and complicated operation, the coagulation of MBF was studied, and applied effect in fruit and vegetable beverages.The equipment of vaccum LHPEF enzymolysis was developed personally, and was used to decompose porcine Hb experiment by Trypsin on the LHPEF. The results showed that the electrical conductivity of porcine Hb was3.45326 x10⁵μs/cm by LHPEF at 37℃,1.88v,200kHz, while the control group was 0.645x10⁴μs/cm. The Hb degradation products by Trypsin on the LHPEF, showed obviously more absorption peak and ribbon than the control group by UV scanning graph, Native-PAGE protein eletrophorogram and HGPC picture.Hemi Peptide-Fe was separated from enzymolysis products by MCAC. The results showed that the extracted Hemi Peptide-Fe showed only one clear ribbon and a characteristic absorption peak through twice chromatography. The molecular mass of Hemi Peptide-Fe was 7529.0968Da, Ferric Content of 10%Hemi Peptide-Fe 1013.5μg/L, theory pl 8.10, proteolipid factor 92.73, GRAVY-0.023,six high hydrophobicity areas and three high hydrophilcity areas, no acrossing film area, suppression ratio of in vitro superoxide anion clearing 81.579%, ratio of-OH clearing 78.3%.The coagulation of Hemi Peptide-Fe by MBF was taken for the convenience of industrial application. Rhodococcus eiythropolis was used to ferment in the cheapness fermentation broth.And the fermentation broth formula was sugar 30g/L, urea7.5g/L, CaCl₂2g/L, MgSO₄ lg/L, NaCI 0.2g/L, pH 9, at 37℃,150r/min,72h, the flocculation rate of MBF produced by fermentation to Kaolin Suspension was 89.85%. It designed 3 factors and 3 levels of secondary orthogonal rotation(23 teams).Three factors were MBF, MBF and (3-cyclodextrin, MBF and chitosan. The recovery rate of Hemi Peptide-Fe was detection of indicators. The results showed that pH6.0,45℃, ultrasonic treatment 15min, flocculation recovery of Hemi Peptide-Fe up to 87.47%. With MBF 4%, (3-dextrin 20%,1.591% chitosan as a composite flocculants, the flocculation recovery of Hemi Peptide-Fe up to 88.47%. MBF flocculation of Hemi Peptide-Fe precipitation was crowding type.The application of adding Hemi Peptide-Fe in vegetable and fruit beverage and animal test were studied. The results showed that the optimal parameters of adding Hemi Peptide-Fe in vegetable and fruit beverage was:Hemi Peptide-Fe lOmg/L, tomato juice 200ml/L, Chinese date juice 200ml/L, liquorice juice 20ml/L, citric acid 0.2g/L, honey 15ml/L, xanthan 0.2g/L, CMCNa 0.4g/L, Sodium Alginate 0.4g/L, and vitamin C 2.0g/L, adjusted to 1Lby pure water. The apparent digestibility of Hemi Peptide-Fe in rate was 83.7%, LD507095mg/kg-bw and 7968-6222mg/kg'bw (P＜O.5). After 12w of feeding femal and male rats with high dosage of Hemi Peptide-Fe, the ATL, ATS, BUN, ALB, Hb and TP are respectively 37.8+2.6 U/L, 154.6±9.4U/L,5.75±0.3mmol/L,34.8±3.0g/L,154.2±4.8g/L,7.4±0.3x10¹² counts/L and 60.3±1.6g/L, compared feeding saline water, the ATL, ATS, BUN, ALB, Hb and TP are 36.6±2.8 U/L,152.3±8.6 U/L,5.85±0.4mmol/L,35.5±2.5g/L,145.6±6.5g/L,7.3±0.6x10¹²counts/L and 61.5±6.4 g/L;the male indexes of ATL, ATS, BUN, ALB, Hb and TP are 36.4±4.0 U/L,154.6±9.6U/L,83±0.3mmol/L,34.8±4.3g/L,153.6±7.5g/L,7.7±0.5x10¹²counts/Land62.6±2.4 g/L, compared feeding saline water, the ATL, ATS, BUN, ALB, Hb and TP of male rats are 37.4±2.6U/L,153.2±10.4U/L,5.88±0.8mmol/L,34.2±3.6g/L,148.9±9.6 g/L,7.5±0.4x1012counts/L and 60.6±45 g/L. The appearance and anatomical characteristics of mouse after acute toxicity test and long term toxicity experiment showed that not significant different discrepancy which compared with control subjects.