本文選題：膠原蛋白肽 + 酶解　； 參考：《山東農(nóng)業(yè)大學(xué)》2017年碩士論文

【摘要】：膠原蛋白肽是以動(dòng)物膠原為原料通過(guò)酶法、酸堿法或熱降解法制備而成的小分子多肽混合物,在功能食品、美容護(hù)膚品、藥物控釋材料和診斷輔料等領(lǐng)域應(yīng)用廣泛。膠原蛋白肽有著抗氧化、防治骨質(zhì)疏松和抑制血管緊張素轉(zhuǎn)化酶(ACE)活性等許多優(yōu)秀的生物活性,因此膠原蛋白肽的制備成為近年的研究熱點(diǎn),具有重要的意義。對(duì)于傳統(tǒng)的膠原蛋白制備技術(shù)而言,其存在分子量范圍寬、工藝條件不完善和降解過(guò)程不可控等缺點(diǎn)存在。本研究以膠原蛋白的不同蛋白酶的傳統(tǒng)酶解反應(yīng)過(guò)程研究為基礎(chǔ),考察了基于超濾膜分離技術(shù)對(duì)于膠原蛋白肽的提取工藝,采用酶解-膜分離集成工藝提取分子量均一可控的的膠原蛋白肽,并采用高效凝膠色譜對(duì)制備的膠原蛋白肽分子量范圍進(jìn)行檢測(cè),以牛明膠為原料探索了分子量均一多肽的制備方法。具體內(nèi)容如下:1.酶解工藝研究:實(shí)驗(yàn)采用牛明膠為原料,利用不同蛋白酶對(duì)其進(jìn)行酶解,探究其酶解動(dòng)力學(xué),對(duì)酶解速率及酶解程度進(jìn)行探索,經(jīng)實(shí)驗(yàn)所得,經(jīng)酶解60min后10k Da以下的小分子多肽所占比例分別為:堿性蛋白酶44.7%、胰蛋白酶39.8%、木瓜蛋白酶35.7%和中性蛋白酶15.3%。2.膠原多肽膜分離研究:膜分離過(guò)程中多肽的透膜行為是進(jìn)行酶解-膜分離集成工藝的基礎(chǔ),實(shí)驗(yàn)研究了明膠酶解產(chǎn)物超濾處理過(guò)程中溶液體積、循環(huán)流量等對(duì)不同分子量多肽跨膜行為的影響。實(shí)驗(yàn)對(duì)1.2L、600m L、300m L的明膠溶液進(jìn)行比較,60min內(nèi)3k Da左右的多肽組分變化量分別為分別為7.84%、16.99%、21.05%。3.酶解-膜分離集成過(guò)程研究:酶解膜分離集成過(guò)程采用連續(xù)酶解反應(yīng),在酶解的過(guò)程中通過(guò)中空纖維膜進(jìn)行多肽過(guò)濾。實(shí)驗(yàn)采用了堿性蛋白酶酶解,使用300m L明膠溶液進(jìn)行循環(huán)反應(yīng)。在特定條件下經(jīng)3k Da中空纖維膜的酶解-膜分離集成反應(yīng)60min后,相對(duì)比傳統(tǒng)酶解反應(yīng)酶解速率提高了15%,所得多肽組分分子量集中在1700與600Da左右。
[Abstract]:Collagen peptide is a small polypeptide mixture prepared from animal collagen by enzymatic, acid-base or thermal degradation. It is widely used in functional foods, cosmetic products, drug controlled release materials and diagnostic excipients. Collagen peptide has many excellent biological activities such as antioxidation, prevention and treatment of osteoporosis and inhibition of angiotensin converting enzyme (ACE) activity. Therefore, the preparation of collagen peptide has become a hot topic in recent years and has important significance. For the traditional collagen preparation technology, there are some disadvantages such as wide molecular weight range, imperfect process conditions and uncontrollable degradation process. Based on the traditional enzymatic hydrolysis process of different proteases of collagen, the extraction process of collagen peptides based on ultrafiltration membrane separation was investigated. Collagen peptides with uniform molecular weight and controllable molecular weight were extracted by the integrated process of enzymatic hydrolysis and membrane separation, and the molecular weight range of the prepared collagen peptides was determined by high performance gel chromatography. The preparation method of homogenous polypeptide with molecular weight was studied by using bovine gelatin as raw material. The details are as follows: 1. Study on enzymatic hydrolysis technology: bovine gelatin was used as raw material to hydrolyze it with different proteases, and the kinetics of enzymatic hydrolysis was studied. The rate and degree of enzymatic hydrolysis were explored. The proportion of small polypeptides less than 10 kDa after enzymatic hydrolysis of 60min were: alkaline protease 44.7, trypsin 39.8, papain 35.7% and neutral protease 15.30.2. Study on the Separation of Collagen Peptide membrane: the permeation behavior of polypeptide in the process of membrane separation is the basis of the integrated process of enzymatic hydrolysis and membrane separation. The volume of solution during ultrafiltration treatment of gelatin hydrolysate was studied experimentally. Effects of cyclic flow rate on the transmembrane behavior of polypeptides with different molecular weight. The changes of polypeptide components about 3kDa in 1.2L ~ 600m / L ~ (300ml) gelatin solution in 60 min were 7.84 ~ 16.995% and 21.05% 路3 ~ (-1), respectively. Studies on the Integrated process of Enzymatic Hydrolysis and membrane Separation: the integrated process of enzymatic membrane separation was carried out by continuous enzymatic hydrolysis and polypeptide filtration was carried out by hollow fiber membrane in the process of enzymatic hydrolysis. The enzymatic hydrolysis of alkaline protease was carried out, and the reaction was carried out by using 300 mL gelatin solution. Under certain conditions, the enzymatic hydrolysis and membrane separation integrated reaction of 60min with 3kDa hollow fiber membrane increased the hydrolysis rate by 15%, and the molecular weight of the peptides was about 1700 and 600Da.
【學(xué)位授予單位】：山東農(nóng)業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：TQ936.16

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