The Preparation Of Heme Chloride By Enzymatic Hy Drolysis Of Yak Hemoglobin And The Structure Of Hemin

In this study, fresh yak blood as the raw material, using the single factor and orthogonal experiment design and response surface analysis of thrombin chloride extraction and purification technology of heme was optimized;By apply the method of acid and alkali purification separation and purification of chlorinated hemoglobin, and extracting the product structure was studied. The results are as follows;1. Papain enzymolysis yak hemoglobin optimum process conditions for the preparation of chlorinated hemoglobin; Enzyme solution temperature of 55 ℃, enzymolysis time 94 min,pH8.0 adding amount of 0.08 g, on this condition chlorinated hemoglobin concentration is6.43 μg/mL.2. Eutral protease enzymolysis yak hemoglobin optimum process conditions for the preparation of chlorinated hemoglobin; Celsius temperature was 51℃,enzyme hydrolysis solution time was 110 min,pH was 7.5,enzyme addition was 0.09 g,under the condition,chlorinated hemoglobin content was 5.87μg/mL.3. Compound protease enzymolysis yak hemoglobin optimum process conditions for the papain enzyme: neutral protease was 4:6, Celsius temperature was 60 ℃,enzyme hydrolysis s olution time was 105 min,pH was 8.0,on this condition,chlorinated hemoglobin content was 6.76 μg/mL.4.Chlorinated hemoglobin product purification conditions as follows,NaOH concentration was 0.5 mol/L,pH was 4.0,acidification let stand for 10 min,under the condition,the chlorinated hemoglobin purity was 87.03%.5. The quantitative actual of chlorinated hemoglobin content was 41.23%, the iron content was 0.2631 g/g measured by atomic absorption spectrometry.6. The infrared spectrum qualitative analysis to extract product standard was consistent with the Sigma company.In conclusion,compared with the single enzyme extraction method, the combination of the yak and the neutral protease was better.The high purity of the chloride can be obtained by the method of acid and alkali purification.The product had high iron content and stab le structure,yak blood has the potential to be an important source of the extraction of the heme.