Research On The Preparation Method Of Casomorphin From Casein With Pepsin

Milk includes the massive protein, its content approximately for 3.3 -3.5% of all milk. Mainly have the casein, lactalbumin,lactoglobulin and some enzyme class has the important physiological function, casein account for 8.8% .Casein includes the eight class aminoacid human body essential dish to plant, is one kind of full rate protein (the PER value is 2.5 ),can provide the essential amino acid for the organism growth. Besides the nutrition function, its peptide section latented biology activeness which produces after the proteinase decomposition has received in the animal digestive tract widely takes.We have this kind the biological function and the physiological action peptide are called the biological activitive peptides. Divides into according to its function: opioid peptide, falls the blood pressure peptide, the anti- thrombus peptide, the immunity promotion peptide, the promotion mineralion absorption peptide, the antibacterial peptide and so on many types.In each biological activity peptides, opioid peptide is studies mostly, divides into the endogene opioid peptide and the extraneous source opioid peptide two big types. The endogene opioid peptide is exists to the human body brain, the nerve ending morphine type matter, they can in vivo synthesis and hand over the nature and the nerve μ,d,κ acceptor mutually affect, in abbreviation the endogene opioid peptide;The extraneous source opioid peptide is exists in extraneous source food, its opioid activeness equally can reverse with the morphine by the natrium Luo alkone, in abbreviation extraneous source opioid peptide. B-Casomorphin-7 is produced after the casein by proteinase is the representative extraneous source opioid peptide. Casomorphin usually is from 6-casein also calls B-CM-7.Since Brantl first separates from the casein hydrolysate to β-Casomorphin-7 in 1979,regarding β-Casomorphin-7's preparation, separation, purification and so on each aspect has widespread on reseach and yields many results. But domestic basically pauses regarding β-Casomorphin-7 research on the structure and function level, only Zhang Yuanshu of Nanjing agricultural college has carried on the preliminary discussion regarding β-Casomorphin-7 preparation craft. This experiment main content was in the former literature report foundation the key detailed research on Preparation method of β-Casomorphin-7 made of Caseinwith pepsin.This experiment mainly is composed by three parts. The first part, the best hydrolisis condition establishment of B-Casomorphin-7 production. Firstly through single factor experimental study substrate concentration, enzyme density, temperature and PH to hydrolisis influence, and based on the difference DH selected the single factor optimum condition. Afterwardr selects four factors three horizontal orthogonals experiments design to carry on to the best enzymolysis further discusses, and obtains the best test condition according to the test result union actual factor. After obtains the optimum condition, continues studies the hydrolisis time with relations of DH, is limited to hydrolisis time 0.5h—3.5h separately prepares sample group of seven .The second part, the establishment condition of sephadex and separation of sample. This part first acts according to the need of experiments t to choose sephadex gel, and choice pepsin enzymolysis sample regarding the luminosity, buffer , the column level altitude, elutes factor and so on speed of flow carries on the comparison and analyzes for establishes the best condition to separation. Under this optimization condition ,put the standard sample carries on with the stable speed elutes, according to suitable elutes the volume to determine the standard curve and carries on the separation experimentation , according to the standard curve judgement the molecular weight, and carries on the goal take the order as the. basis to elute thepeak the collection.The third part, examination of B-Casomorphin-7. The experiment has first carried on the chromatograph condition exploration, including the choice of separation column, flows as well as the column temperature column pressure, the speed of flow, the examination wave length and the sensitivity choice, uses the chromatograph external standard law in the establishment standard analysis method foundation to prepare the dish -sample for front to carry on the examination. Comparisoning standard retention time and the peak area, analysis examination result, and quantitative evaluation B-Casomorphin-7.