本文選題：植物甾醇酯 + 酶法合成��； 參考：《哈爾濱商業(yè)大學(xué)》2017年碩士論文

【摘要】：植物甾醇是一種具有生物活性的物質(zhì),由于水溶性和脂溶性都不是很好,限制了其應(yīng)用。通過(guò)酶催化的方式,與油酸反應(yīng)生成植物甾醇油酸酯,它不僅具有植物甾醇與脂肪酸兩部分的營(yíng)養(yǎng)價(jià)值,而且改善了植物甾醇的脂溶性和降膽固醇效果,應(yīng)用范圍更廣。大豆油脫臭餾出物(SODD)是植物油精煉過(guò)程中的副產(chǎn)物,富含游離脂肪酸、植物甾醇、天然維生素E等具有應(yīng)用價(jià)值的物質(zhì)。因此從大豆油脫臭餾出物中提取出植物甾醇和脂肪酸,并通過(guò)酶法合成植物甾醇脂肪酸酯具有重大意義。本實(shí)驗(yàn)構(gòu)建了在非水相中酶法合成植物甾醇酯的反應(yīng)體系,優(yōu)化了合成工藝,并對(duì)催化酶進(jìn)行了固定化研究。從大豆油脫臭餾出物中提取出植物甾醇和脂肪酸,根據(jù)優(yōu)化的工藝酶法合成植物甾醇脂肪酸酯,并進(jìn)行了定性分析。主要研究?jī)?nèi)容如下:(1)在非水相體系中進(jìn)行酶促反應(yīng),合成植物甾醇酯。分別篩選脂肪酶和有機(jī)溶劑介質(zhì),實(shí)驗(yàn)結(jié)果表明,皺褶假絲酵母(Candida rugosa)脂肪酶催化合成植物甾醇酯的酯化率最高;在正己烷中,酶法合成植物甾醇酯的酯化率效果最好。確定了酯化反應(yīng)的反應(yīng)體系后,對(duì)反應(yīng)產(chǎn)物進(jìn)行了薄層色譜分析,在薄層色譜中有新物質(zhì)生成。對(duì)分離純化后的產(chǎn)物進(jìn)行紅外光譜分析,在1734cm-1處有明顯吸收峰,是酯鍵的特征吸收峰。因此證明反應(yīng)合成的產(chǎn)物為植物甾醇油酸酯。(2)研究了影響酯化反應(yīng)的因素,對(duì)油酸與植物甾醇的物質(zhì)的量的比,酶用量,酯化的溫度和時(shí)間4個(gè)因素進(jìn)行了探討。經(jīng)單因素實(shí)驗(yàn)考察后,通過(guò)甾醇酯化率的響應(yīng)曲面分析,優(yōu)化了反應(yīng)條件:底物摩爾比(油酸/植物甾醇)為3:1,酶用量為底物總質(zhì)量的7.2%,反應(yīng)溫度為45.3℃,反應(yīng)時(shí)間為25.3h,最優(yōu)條件下酯化率可達(dá)71.95%。(3)采用物理吸附法對(duì)CRL脂肪酶(Candida rugosa lipase)進(jìn)行固定化,以植物甾醇轉(zhuǎn)化率和酶活力為考察指標(biāo),實(shí)驗(yàn)表明:CRL脂肪酶分別在磷酸緩沖溶液中和有機(jī)溶劑正己烷中固定化,前者效果更好;CRL脂肪酶在磷酸緩沖溶液中分別以幾種不同載體固定化,進(jìn)行批式反應(yīng),對(duì)幾種不同固定化酶載體進(jìn)行篩選,大孔吸附樹(shù)脂HP-20操作穩(wěn)定性最好,性能最優(yōu)。連續(xù)反應(yīng)3次后,酶活力為509.92U/g,酯化反應(yīng)的酯化率為62.99%,仍有較高的催化能力。(4)研究了溶劑冷卻結(jié)晶法從大豆油脫臭餾出物中提取出植物甾醇,實(shí)驗(yàn)確定最佳提取工藝:丙酮-甲醇混合溶劑的比例為1:2,溶劑原料比為3:1(V/W),養(yǎng)晶時(shí)間為18h,洗滌粗甾醇的溶劑為正己烷,洗滌粗甾醇的溫度為25℃,洗滌粗甾醇的時(shí)間為15min。然后從提取植物甾醇后的濾液中提取出脂肪酸,根據(jù)前三章優(yōu)化好的合成工藝酶法合成植物甾醇酯。通過(guò)薄層色譜法鑒定分析,在色譜中有新的物質(zhì)出現(xiàn),可初步判定是植物甾醇脂肪酸酯。再通過(guò)紅外光譜法表征,經(jīng)紅外圖譜分析,圖中在1737cm-1處有明顯的伸縮振動(dòng)吸收峰,是酯鍵的特征吸收峰,因此證明了從大豆油脫臭餾出物中提取的植物甾醇和脂肪酸,在非水相中經(jīng)脂肪酶催化合成的產(chǎn)物為植物甾醇脂肪酸酯。
[Abstract]:Phytosterol is a bioactive substance. Because of its water solubility and fat solubility are not very good, it restricts its application. The plant sterol oleate is produced by enzyme catalyzed reaction with oleic acid. It not only has the nutritional value of two parts of plant sterols and fatty acids, but also improves the fat solubility and cholesterol lowering of plant sterols. Soybean oil deodorization distillate (SODD) is a by-product of plant oil refining, which is rich in free fatty acids, phytosterols, natural vitamin E and other substances. Plant sterols and fatty acids are extracted from soybean oil deodorizer distillate, and phytosterol fatty acid esters are synthesized by enzyme method. In this experiment, the reaction system of phytosterol ester synthesis in non aqueous phase was constructed, the synthesis process was optimized, and the catalytic enzyme was immobilized. Plant sterol and fatty acid were extracted from the deodorizer distillate of soybean oil, and the phytosterol fatty acid ester was synthesized according to the optimized process enzyme method, and the qualitative analysis was carried out. The main contents are as follows: (1) enzymatic reaction in non aqueous phase and synthesis of phytosterol esters. Lipase and organic solvent medium are screened respectively. The results show that the esterification rate of phytosterol esters catalyzed by Candida rugosa lipase catalyzed by crease Candida folds is the highest; esterification efficiency of phytosterol ester in n-hexane After determining the reaction system of esterification, the reaction product was analyzed by TLC, and the new substance was formed in the thin layer chromatography. The infrared spectrum analysis of the purified product was carried out by the infrared spectrum analysis. The absorption peak of the ester bond was found at 1734cm-1, and the product of the reaction was phytosterol oleic acid. (2) the factors affecting the esterification were studied. The ratio of the amount of oleic acid to phytosterol, the amount of the enzyme, the temperature and the time of the esterification were discussed. After a single factor experiment, the reaction strip was optimized by the response surface analysis of sterol esterification rate: the molar ratio of the substrate (oleic acid / phytosterol) was 3:1, the amount of the enzyme was The total mass of the substrate is 7.2%, the reaction temperature is 45.3, the reaction time is 25.3h, the esterification rate can reach 71.95%. (3) under the optimal condition. The physical adsorption method is used to immobilize the CRL lipase (Candida rugosa lipase), and the conversion rate of phytosterol and the activity of the enzyme are considered as the indexes. The results show that the CRL lipase is neutralized in the phosphate buffer solution and the organic matter respectively. The former is immobilized in n-hexane, the former is better, CRL lipase is immobilized on several different carriers in the phosphate buffer solution, batch reaction is carried out, and several different immobilized enzyme carriers are screened. The macroporous adsorption resin HP-20 has the best stability and the best performance. After 3 times the continuous reaction, the enzyme activity is 509.92U/g and esterification reaction. The esterification rate is 62.99% and still has high catalytic ability. (4) the extraction of phytosterol from the deodorizing distillate of soybean oil is studied by the solvent cooling crystallization method. The optimum extraction process is determined by the experiment: the proportion of acetone methanol mixed solvent is 1:2, the ratio of solvent to raw material is 3:1 (V/W), the crystallization time is 18h, the solvent washing crude sterol is n-hexane, and the washing is coarse. The temperature of sterol is 25 degrees C, the time of washing crude sterol is 15min. and then the fatty acid is extracted from the filtrate of phytosterol extraction. The plant sterol ester is synthesized by the optimized synthetic process enzyme method based on the first three chapters. A new substance in the chromatogram is identified and analyzed by TLC, and the phytosterol fatty acid ester can be preliminarily determined. It was characterized by infrared spectroscopy. After the analysis of infrared spectrum, there was an obvious absorption peak of expansion and vibration at 1737cm-1, which was the characteristic absorption peak of ester bond. Therefore, phytosterol and fatty acids extracted from soybean oil deodorizer distillate were proved to be phytosterol fatty acid esters in non aqueous phase.

【學(xué)位授予單位】：哈爾濱商業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：TS202.1

【參考文獻(xiàn)】

相關(guān)期刊論文 前10條

1 趙巖;唐國(guó)勝;侯瑩瑩;牛淑靜;郜玉鋼;韓雪;張悅怡;沈彥龍;張連學(xué);;植物甾醇酯合成工藝研究進(jìn)展[J];食品安全質(zhì)量檢測(cè)學(xué)報(bào);2015年02期

2 許青青;金文彬;蘇寶根;楊亦文;任其龍;;植物甾醇酯的化學(xué)合成及其分離研究進(jìn)展[J];中國(guó)糧油學(xué)報(bào);2014年03期

3 蔣振華;于敏;任立偉;周華;韋萍;;有機(jī)相中固定化脂肪酶催化合成植物甾醇酯[J];催化學(xué)報(bào);2013年12期

4 張蕾;陳慶森;閻亞麗;龐廣昌;;植物甾醇對(duì)心血管疾病作用的研究現(xiàn)狀[J];食品科學(xué);2013年23期

5 左玉;;植物甾醇研究與應(yīng)用[J];糧食與油脂;2012年07期

6 董玲;鄭明明;郭萍梅;李文林;黃鳳洪;;酶法合成植物甾醇酯工藝的研究進(jìn)展[J];中國(guó)油脂;2012年06期

7 姜紹通;鄭海杰;龐敏;;植物甾醇酯的脂肪酶促催化合成及其分析表征[J];食品科學(xué);2012年06期

8 馬玲芝;錢(qián)俊青;;離子交換樹(shù)脂固定化脂肪酶催化合成蔗糖乙酯[J];浙江工業(yè)大學(xué)學(xué)報(bào);2012年01期

9 于殿宇;王騰宇;陳昊;朱秀清;江連洲;胡立志;;生物酶法合成植物甾醇酯的研究[J];食品工業(yè)科技;2011年06期

10 張琛;張寬朝;章琛;繆偉;;海藻酸鈉包埋法制備固定化菠蘿蛋白酶[J];生物加工過(guò)程;2009年06期

相關(guān)碩士學(xué)位論文 前8條

1 阮慧娜;植物甾醇酯的新型制備方法研究[D];浙江大學(xué);2014年

2 朱振南;菜籽油脫臭餾出物的維生素E和甾醇提取及共軛亞油酸甾醇酯的制備[D];華中科技大學(xué);2013年

3 劉虹蕾;脂肪酶非水相合成植物甾醇酯及其動(dòng)力學(xué)研究[D];江南大學(xué);2012年

4 李江濤;脂肪酶催化合成α-亞麻酸植物甾醇酯的研究[D];華中農(nóng)業(yè)大學(xué);2008年

5 李奇;大豆油脫臭餾出物中生育酚、甾醇和脂肪酸的提取工藝研究[D];天津大學(xué);2008年

6 黃琴;植物甾醇酯的合成及活性研究[D];湖北工業(yè)大學(xué);2006年

7 沈斌;木瓜蛋白酶的柔性固定化研究[D];南京工業(yè)大學(xué);2004年

8 韋繼慷;植物甾醇提取工藝研究[D];浙江大學(xué);2002年

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