本文選題：牛皮邊角料 + 膠原�。� 參考：《鄭州大學(xué)》2017年碩士論文

【摘要】：作為動(dòng)物體內(nèi)含量最多、分布最廣的一種蛋白質(zhì),膠原具有生物相容性好、可降解、免疫原性低等優(yōu)點(diǎn),可應(yīng)用于生物醫(yī)藥、食品、化妝品等諸多領(lǐng)域。膠原的應(yīng)用與其分子量等息息相關(guān)。本文以牛皮邊角料為原料,首先對(duì)原料進(jìn)行預(yù)處理達(dá)到除雜和脫毛的目的。隨后,利用單因素實(shí)驗(yàn)探索了胃蛋白酶酶解牛皮提取膠原的工藝,以膠原提取率為參數(shù)分析了料液比(牛皮質(zhì)量與0.5 mol/L醋酸溶液體積比)、酶用量、溫度、初始pH和酶解時(shí)間五個(gè)變量對(duì)提取膠原的影響。借助傅立葉紅外光譜(FTIR)和掃描電子顯微鏡(SEM)表征了所提取膠原的結(jié)構(gòu)與形貌。借助聚丙烯酰胺凝膠電泳(SDS-PAGE)研究了不同條件下提取膠原的分子量分布,借助紫外可見分光光度計(jì)(UV-vis)在波長(zhǎng)為313 nm下研究了所提取膠原的體外成纖能力,并利用差示掃描量熱法(DSC)表征了其熱變性行為,初步建立了從牛皮中提取膠原的條件與膠原主要性能的關(guān)系,為膠原的提取和應(yīng)用提供了一定的理論依據(jù)。預(yù)處理后的牛皮檢測(cè)結(jié)果表明:預(yù)處理對(duì)牛皮固含量的影響較小。牛皮pH處于中性范圍,熱變性溫度與文獻(xiàn)報(bào)道中相符合,為50.3°C。單因素實(shí)驗(yàn)表明:其它條件一定時(shí),僅料液比改變,隨醋酸溶液體積的增加,膠原提取率逐漸提高;酶用量增大時(shí),膠原提取率也隨之增大,酶用量從1.5%到2.5%時(shí)提取率提高最為顯著;溫度對(duì)膠原的提取影響突出,溫度過高會(huì)導(dǎo)致提取率顯著下降;初始pH低時(shí)膠原提取率偏低,但當(dāng)pH增大到2.0以上,提取率在設(shè)定pH梯度范圍內(nèi)所受影響微弱;酶解時(shí)間得到的結(jié)果是,隨酶解時(shí)間的延長(zhǎng),膠原提取率呈遞增趨勢(shì),但酶解達(dá)到一定時(shí)間后,很難通過改變酶解時(shí)間顯著提高其提取率。對(duì)所提取試樣的紅外光譜、數(shù)碼照片和掃描電鏡圖研究表明:得到的產(chǎn)物為鮮亮純白的膠原,呈片層狀結(jié)構(gòu)。提取條件對(duì)所提取膠原結(jié)構(gòu)影響不大。由不同條件下提取出膠原的電泳結(jié)果可知:膠原的分子量大小及分布受料液比、酶用量、溶液初始pH和酶解時(shí)間影響較小,所有電泳圖都存在α條帶且絕大多數(shù)條帶在66.2 kDa以上;提取溫度改變會(huì)使膠原分子量發(fā)生顯著變化,50°C時(shí)膠原的α條帶消失,分子量分布更寬,因此溫度是控制膠原分子量的一個(gè)關(guān)鍵因素。與煮沸的膠原試樣對(duì)比發(fā)現(xiàn),料液比、酶用量、溶液初始pH和酶解時(shí)間四種因素條件下得到的膠原均具有體外成纖能力,其體外成纖能力強(qiáng)弱略有差異;在不同的溫度下提取膠原的體外成纖能力方面,僅50°C條件下提取的膠原喪失了體外成纖能力。DSC結(jié)果可知:膠原的熱變性溫度整體在45°C以上。酶用量、初始pH和酶解時(shí)間對(duì)膠原變性溫度影響不大;但牛皮質(zhì)量一定時(shí),隨醋酸溶液體積增大,膠原熱變性溫度降低;當(dāng)溫度低于37°C時(shí),膠原的熱變性溫度受提取溫度影響較小,當(dāng)溫度高于37°C后膠原的熱變性溫度降到50°C以下。
[Abstract]:As one of the most abundant and widely distributed proteins in animals, collagen has the advantages of good biocompatibility, biodegradability and low immunogenicity. It can be used in many fields such as biomedicine, food, cosmetics and so on. The application of collagen is closely related to its molecular weight. In this paper, the raw material is used as raw material, the raw material is pretreated to achieve the purpose of removing impurity and depilation. Then, the process of extracting collagen from bovine skin by pepsin enzymatic hydrolysis was studied by single factor experiment. The ratio of material to liquid (the volume ratio of bovine skin to 0.5 mol / L acetic acid solution), the amount of enzyme and the temperature were analyzed based on the extraction rate of collagen. Effects of initial pH and enzymatic hydrolysis time on collagen extraction. The structure and morphology of the extracted collagen were characterized by Fourier transform infrared spectroscopy (FTIR) and scanning electron microscope (SEM). The molecular weight distribution of collagen extracted under different conditions was studied by using polyacrylamide gel electrophoresis (SDS-PAGE), and the in vitro fibrinogenic ability of the extracted collagen was studied by UV-vis spectrophotometer (UV-vis) at the wavelength of 313 nm. The thermal denaturation behavior was characterized by differential scanning calorimetry (DSC). The relationship between the extraction conditions and the main properties of collagen was preliminarily established, which provided a theoretical basis for the extraction and application of collagen. The results showed that pretreatment had little effect on the solid content of leather. The pH of the skin is in the neutral range, and the thermal denaturation temperature is consistent with that reported in the literature, which is 50.3 擄C. The results of single factor experiments showed that the extraction rate of collagen increased with the increase of the volume of acetic acid solution, and the extraction rate of collagen increased with the increase of enzyme dosage. The extraction rate of collagen increased most significantly when the enzyme dosage was from 1.5% to 2.5. The effect of temperature on the extraction of collagen was obvious, and the extraction rate decreased significantly when the temperature was too high. The extraction rate of collagen was low when the initial pH was low, but when pH increased to more than 2.0, The extraction rate is weak in the range of pH gradient, the result of enzymatic hydrolysis time is that the extraction rate of collagen increases with the extension of enzymatic hydrolysis time, but when the enzymatic hydrolysis reaches a certain time, the extraction rate of collagen increases with the increase of hydrolysis time. It is difficult to improve the extraction rate by changing the enzymatic hydrolysis time. The infrared spectra, digital photographs and scanning electron microscopy of the extracted samples showed that the obtained products were pure white collagen with lamellar structure. The extraction conditions had little effect on the structure of collagen extracted. The electrophoretic results of collagen extracted under different conditions showed that the molecular weight and distribution of collagen were less affected by the ratio of material to liquid, the amount of enzyme, the initial pH of solution and the time of enzymatic hydrolysis. There were 偽 bands in all electrophoretic patterns and most of them were above 66.2 kDa. The molecular weight of collagen disappeared and the molecular weight distribution was wider when the molecular weight of collagen disappeared at 50 擄C when the extraction temperature changed significantly. Therefore, temperature is a key factor to control the molecular weight of collagen. Compared with boiled collagen samples, the collagen obtained under four conditions: the ratio of material to liquid, the amount of enzyme, the initial pH of solution and the time of enzymatic hydrolysis all had the ability to form fiber in vitro, and the ability of fibrin formation in vitro was slightly different. The results of DSC showed that the thermal denaturation temperature of collagen was above 45 擄C. The amount of enzyme, initial pH and enzymatic hydrolysis time had little effect on the denaturation temperature of collagen, but the thermal denaturation temperature of collagen decreased with the increase of acetic acid solution volume when the weight of the skin was constant, and when the temperature was lower than 37 擄C, the denaturation temperature of collagen decreased. The thermal denaturation temperature of collagen was less affected by the extraction temperature. When the temperature was higher than 37 擄C, the thermal denaturation temperature of collagen decreased below 50 擄C.
【學(xué)位授予單位】：鄭州大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：TQ936

【相似文獻(xiàn)】

相關(guān)期刊論文 前10條

1 趙敏;;一種寬分子量分布氟橡膠的制備方法[J];橡膠工業(yè);2010年04期

2 丁正友;周蘊(yùn);高翔;;腈綸的分子量和分子量分布與腈綸工藝[J];金山石油化工化纖技術(shù);1984年04期

3 張志英;用高速凝膠色譜法測(cè)定硝化棉的分子量及分子量分布[J];消防科技;1986年03期

4 寶凈生,韓寶珍,徐朝儔;聚對(duì)苯二甲酰對(duì)苯二胺的分子量分布[J];產(chǎn)業(yè)用紡織品;1988年S1期

5 黃榮南,錢蓉娣;丁腈軟膠的分子量及分子量分布的測(cè)試[J];蘭化科技;1989年Z1期

6 ;用沉淀分級(jí)法測(cè)定三醋酸纖維素的分子量分布[J];北京化工大學(xué)學(xué)報(bào)(自然科學(xué)版);1976年01期

7 ;用硅凝膠滲透色譜柱測(cè)定三醋酸纖維素的分子量分布[J];北京化工大學(xué)學(xué)報(bào)(自然科學(xué)版);1976年01期

8 謝洪泉,汪月生;從分子量分布探，

本文編號(hào)：2056595