植物甾醇酯的酶法合成

以油酸和植物甾醇为反应底物,利用脂肪酶催化合成植物甾醇油酸酯。对脂肪酶和反应介质进行了筛选,同时对影响合成植物甾醇油酸酯反应的因素进行了探究,考察了底物摩尔比(油酸甾醇比)、酶添加量(脂肪酶质量/底物总质量)、反应温度及反应时间对酯化反应的影响。在单因素的基础上,经酯化率响应曲面实验优化反应条件,确定最佳反应条件:催化酯化反应的酶为柱状假丝酵母脂肪酶(CRL,Candida rugosa lipase),反应介质为正己烷,底物摩尔比为3∶1,酶添加量为7.2%,反应温度为45.3℃,反应时间为25.3h,验证试验酯化率可达72.95%。     

维生素E琥珀酸酯的酶促合成及优化

在有机溶剂中以维生素E和琥珀酸酐为底物在脂肪酶的催化下合成了维生素E琥珀酸酯。首先对酶促反应的脂肪酶、反应介质和反应温度进行了考察,在所选的几种脂肪酶中,假丝酵母脂肪酶(Candidasp.)的催化活性最好;叔丁醇和DMSO组成的混合溶剂(体积比为2∶3)为最合适的反应介质;30℃为适宜的反应温度。并采用Box-Behnken实验设计和响应面因子分析法对底物摩尔比等其他反应条件进行了优化。维生素E琥珀酸酯最优反应条件为:维生素E浓度0.26mmol.ml-1,维生素E和琥珀酸酐摩尔比1∶5,在5ml叔丁醇和DMSO混合溶剂(体积比为2∶3)中,30℃下在0.02gCandidasp.脂肪酶的催化下反应71h,维生素E琥珀酸酯产率达到98.71%。     

酶促酯化拆分-化学消旋串联法制备S-布洛芬

以消旋布洛芬为反应底物，探究了酶促酯化拆分布洛芬的工艺。利用南极假丝酵母脂肪酶B的对映体选择性催化布洛芬选择性酯化，并对醇的种类、反应介质、底物摩尔比、酶量、干燥剂种类和添加量、温度及时间等反应条件进行了优化，以56%的收率得到了光学纯的S-布洛芬，接下来对酯化产物进行化学法消旋。以二甲亚砜为反应介质，利用二异丙基氨基锂催化布洛芬发生外消旋反应，完全消旋后再进行酶促拆分仍能达到之前的效果。如此循环7次布洛芬的利用率可达到90%以上。     

固定化假丝酵母脂肪酶合成棕榈酸异辛酯

开发了固定化假丝酵母脂肪酶99-125合成棕榈酸异辛酯的工艺.对反应温度、酶用量、底物摩尔比等酯化反应条件进行了研究.脂肪酶以吸附的形式固定在织物膜上,以1g棕榈酸、0.67g异辛醇、0.12g固定化脂肪酶和5ml石油醚组成的反应系统在40℃条件下反应24h,酯化率可达96.6%.固定化酶连续反应9批后酯化率仍维持在90%以上.     

环状芽孢杆菌胞壁溶解酶用于红发夫酵母虾青素提取的研究

采用单因素及均匀实验设计确定了将环状芽孢杆菌胞壁溶解酶用于红发夫酵母破壁提取虾青素的最佳酶作用条件,即粗酶液pH值为5.0,体积为33mL(酶量1603.8U(g(1干酵母),温度37℃,100r(min(1振荡反应16.5h,虾青素的提取率可达到98%以上.通过对不同方法提取所得虾青素的光、热稳定性进行测试,发现酶法提取的虾青素不论是在光照还是在加热条件下均比机械法及酸法破壁提取所得的虾青素稳定.     

法夫酵母生物法生产虾青素的研究进展

介绍了虾青素在功能饲料、食品、化妆品和医药等方面的应用前景以及虾青素的合成方法。在微生物合成中,重点介绍了法夫酵母生产虾青素的方法,阐述了通过基因工程培养、诱变及发酵工艺优化等方法来提高法夫酵母生物合成虾青素的新进展。     

脂肪酶催化合成维生素A酯

研究了有机溶剂中脂肪酶催化合成维生素A酯。对催化合成维生素A棕榈酸酯反应的脂肪酶和反应介质进行了比较,同时对影响合成维生素A棕榈酸酯反应的因素(温度、初始水含量、底物摩尔比、反应时间和酶量等)进行了探讨,优化了反应条件:在10mL不含水分的正己烷中,0.200g维生素A醋酸酯和0.468g棕榈酸在酶量为10%(指固定化酶与维生素A醋酸酯的质量比)的固定化脂肪酶催化下,在30℃、190r/min下反应6h,转化率可以达到75%,固定化酶可连续使用6次以上。     

细胞生长、葡萄糖代谢及副产物重新利用对法夫酵母虾青素合成的影响分析

以两株产虾青素法夫酵母菌株为研究对象,分别在不同摇床转速、初始葡萄糖浓度、乙醇的存在与否等培养条件下,考察葡萄糖浓度、添加乙醇及溶氧状况对法夫酵母细胞生长、代谢副产物产生的影响及其与虾青素合成之间的关系,进一步阐明法夫酵母虾青素合成的机理。实验结果表明:法夫酵母JMU-VDL668菌株的生长速率、葡萄糖代谢速率和乙醇积累量均高于法夫酵母JMU-MVP14菌株,但前者的虾青素合成速率却远低于后者;当发酵液中的葡萄糖含量降低时,酵母细胞对糖代谢副产物进行二次利用,导致有利于虾青素合成的情况出现,虾青素产量得到进一步提高;在初始培养基中添加1 g?L?1乙醇能够在一定程度上促进虾青素的合成。     

叔丁醇体系中脂肪酶催化合成蔗糖乙酸酯

以蔗糖和乙酸乙烯酯为底物,脂肪酶催化合成蔗糖乙酸酯并分离得到了三氯蔗糖生产的关键中间体蔗糖-6-乙酸酯。今对催化合成蔗糖乙酸酯的反应溶剂进行比较,确定叔丁醇为较好的反应介质,详细考察了各反应参数:酶种类、初始水活度、底物摩尔比、酶用量、反应温度和时间对酯化反应效果的影响,确定了该反应体系的最优工艺条件:蔗糖40mmol?L?1,乙酸乙烯酯/蔗糖(mol/mol)=5,初始水活度aw=0.75,温度60℃,20%(w/w,酶/蔗糖)固定化脂肪酶LipozymeTLIM催化,反应24h后酯化率达84.70%。通过硅胶柱层析分离得到高纯度的其中一种产物,经二维核磁(2D-NMR)结构分析后,确定其为蔗糖-6-乙酸酯,占总酯化产物的质量达48.30%。     

红发夫酵母细胞内蛋白质、脂肪酸与虾青素合成关系的研究

为研究红发夫酵母细胞内虾青素合成与蛋白质和脂肪酸代谢之间的关系,实验测定了发酵过程中蛋白质、脂肪酸和虾青素含量的变化。实验结果表明:红发夫酵母细胞内蛋白质的含量在整个发酵周期内变化非常显著,在0～16h(延滞期)和58～76h(指数生长后期)上升,而在16～58h(指数期)和76～124h(稳定期后期)下降。脂肪酸的含量变化趋势与蛋白质的基本相似,但是在稳定期表现出先快速上升然后迅速下降的变化特征。相比之下,虾青素含量的变化规律与蛋白质和脂肪酸相反。上述变化规律表明红发夫酵母细胞内虾青素的合成与蛋白质和脂肪酸的代谢密切相关。在延滞期和稳定前期蛋白质和脂肪酸的快速合成会减少流向虾青素合成方向的碳通量,使虾青素含量减少;而在稳定期后期底物耗尽出现碳饥饿时,蛋白质和脂肪酸的降解又为虾青素的进一步合成提供了碳骨架,提高了虾青素的含量。     

Preparation of alkenyl succinic anhydrides from vegetable oil FAME

Sunflower oil and oleic motif-enriched sunflower oil methyl esters were used to prepare alkenyl succinic anhydrides. A classic batch reactor was selected to carry out the synthesis. The range in which the temperature, reaction time, and molar ratio between the number of moles of maleic anhydride and the equivalent number of double bonds present in the unsaturated vegetable oil methyl esters (the most influential factors in the process) varied was determined in a preliminary study. A secondorder Doehlert uniform network design was used to investigate the influence of the temperature and molar ratio for all the methyl esters on the yield of alkenyl succinic anhydride from methyl oleate, the conversion of methyl oleate, the formation of side reaction products, the Gardner color of the product, and viscosity. The optimal reaction conditions for obtaining the maximal yield (around 95%) of alkenyl succinic anhydride from methyl oleate were 235°C, a molar ratio of 1.5, and a reaction lasting 8 h. However, the products synthesized under these conditions showed high viscosity (215 cP), a very dark color (18+ Gardner color), and a high content of undesirable side products (4%), which hindered their direct industrial use. The increase in the product viscosity was probably due to the formation of side reaction products. A molar ratio of less than 1.5 led to a less viscous product, although with a lower alkenyl succinic anhydride content.     

Production of alkenyl succinic anhydrides from low‐erucic and low‐linolenic rapeseed oil methyl esters

Fatty acid methyl esters from low‐erucic and low‐linolenic rapeseed oil were used to produce alkenyl succinic anhydrides. A second‐order Doehlert uniform network design was used to investigate the influence of the reaction temperature and the molar ratio between the maleic anhydride and the main unsaturated rapeseed oil methyl esters on the yield of alkenyl succinic anhydride from methyl oleate. Further subjects of investigation were the conversion of methyl oleate, the formation of side reaction products, the Gardner color of the product and its viscosity, and finally the content of maleic anhydride remaining in the medium after the reaction. Alkenyl succinic anhydride from methyl oleate was isolated by column chromatography and analyzed by IR, ¹H‐ and ¹³C‐NMR and MS. The optimal reaction conditions for obtaining the maximum yield of alkenyl succinic anhydride from methyl oleate in the experimental domain (80%) were 210‐220 °C and a maleic anhydride/rapeseed oil methyl ester molar ratio of 1.5. However, the products synthesized in these conditions showed a high degree of viscosity (0.45 kg m^?1 s^?1), a very dark color (18 Gardner color) and a high content of undesirable side products (6%), which could hinder their industrial use. A molar ratio of less than 1.5 led to a clearer and less viscous product, although with a lower alkenyl succinic anhydride content.     

Synthesis of starch esters in ionic liquids

Chemical modification of corn starches with succinic anhydride or acetic anhydride was carried out using 1‐butyl‐3‐methylimidazolium chloride (BMIMCl) as a reaction medium. The reaction progress was followed in terms of the degree of substitution (DS) for the starch derivatives. The results showed that the homogeneous esterification of starch at 5 : 1 molar ratio of anhydride/anhydroglucose units at 100°C led to formation of acetates with DS ranging from 0.37 to 2.35 and succinates with DS ranging from 0.03 to 0.93. Moreover, the reaction media applied could be easily recycled and reused. Further, the formation of starch esters was confirmed by the presence of the carbonyl signal in the FTIR and NMR spectra. It was shown that the starch granules were mostly converted from their crystalline structure into amorphous state in the ionic liquid system under the given reaction conditions. © 2009 Wiley Periodicals, Inc. J Appl Polym Sci, 2010     

Synthesis of Vitamin A Esters by Immobilized Candida sp. Lipase in Organic Media

Vitamin A ester was synthesized in organic solvents with immobilized lipase from Candida sp. The types of lipases, influences of solvent, the molar ratio of substrates, the reaction temperature and the water activity in the reaction were studied in detail in order to obtain the optimum conditions for Vitamin A palmitate synthesis. In a system of hexane, 100mg immobilized Candida sp. lipase was used in the presence of 1.2mmol vitamin A acetate and 3.6mmol palmitic acid. The yield of vitamin A palmitate reached 81% in 12h at 25℃. The immobilized Candida sp. lipase was prepared by adsorbing Cand/da sp. fermentation broth on pretreated textile and could be reused for at least six batches.     

Enzymatic synthesis of astaxanthin n-octanoic acid esters

We examined the enzymatic synthesis of astaxanthin n-octanoic acid esters. Carriers for the immobilized enzyme and reaction conditions such as water content, reaction temperature, and time were examined using Candida cylindracea lipase (Lipase OF). Lipase OF) immobilized by a hydrophobic anion exchange resin (10% w/w content of lipase) gave the best yield in the esterification reaction of astaxanthin. Two milligrams of astaxanthin per 750 microL tri-n-octanoin (ca. 0.3%) was optimum because of the low solubility of tri-n-octanoin. The esters were obtained in a yield of 36.4% under the optimal reaction conditions.     

单过氧丁二酸氧化环己酮合成ε-己内酯(英文)

In the absence of catalyst, 70% hydrogen peroxide was used to oxidize succinic anhydride to solid monoperoxysuccinic acid（PSA）. Then PSA was applied to synthesis of ε-caprolactone（ε-CL） by oxidation of cyclohexanone in the heterogeneous system. In order to achieve material recycle, solid precipitated in the process of synthesizing ε-CL was dehydrated via reactive distillation followed by recrystallization to prepare succinic anhydride, which was characterized by IR（infrared spectra） and1HNMR（1H nuclear magnetic resonance）. Effects of molar ratio of PSA to cyclohexanone, acetic acid dosage, reaction temperature, reaction time on conversion of cyclohexanone, yield and selectivity of ε-CL were investigated respectively. The results indicated that conversion of cyclohexanone, yield and selectivity of ε-CL were upto 98.1%, 97.5% and 99.4% respectively under the optimal conditions. In addition, in the process of synthesizing succinic anhydride, the optimal yield of succinic anhydride reached 67.4%.     

Enzymatic production of fatty acid alkyl esters with a lipase preparation from Candida sp. 99‐125

A lipase preparation developed from Candida sp. 99‐125 was used for fatty acid alkyl ester synthesis by both enzymatic esterification of fatty acids, and transesterification of oils and fats. The lipase preparation was chosen based on screening of lipases from commercial sources as well as those produced in the laboratory. The effects of enzyme dosage, solvent types, water absorbent additions, inhibition of short‐chain alcohols, alcohol and acid types, molar ratio of substrates, and reusability of the lipase preparation in esterification were studied. Degree of esterification between oleic acid and methanol under optimal conditions reached 92%. Purity of the methyl ester after washing with water and distillation was 98%. Half‐life of the lipase preparation was calculated to be approximately 340 h. For transesterification of rapeseed oil with the same lipase preparation, the amount of methanol and mode of methanol addition to the reaction were also conducted. Transesterification of the oil with stepwise methanol addition reached 83% after 36 h reaction time.     

Studies on the enzymatic synthesis of lipophilic derivatives of natural antioxidants

The esterification of some natural antioxidants such as cinnamic acid derivatives and ascorbic acid in non-aqueous media, catalyzed by immobilized lipases from Candida antarctica and Rhizomucor miehei, was investigated. The alcohol chain length affected the rate of esterification of cinnamic acids by both lipases. Higher reaction rates were observed when the esterification was carried out with medium- or long-chain alcohols. The rate also depended on aromatic acid structure. The reactivity of the carboxylic function of the cinnamic acids was affected by electron-donating substituents in the aromatic ring. Higher yields were observed for the esterification of p-hydroxyphenylacetic acid (97%) catalyzed by C. antarctica lipase and for the esterification of cinnamic acid (59%) catalyzed by R. miehei lipase. Candida antarctica lipase was more suitable for producing ascorbic acid fatty esters, catalyzing with a relatively high yield (up to 65% within 24 h) the regioselective esterification of ascorbic acid with various fatty acids in 2-methyl-2-propanol. The reaction rate and yield depended on the fatty acid chain length and on the molar ratio of reactants. All ascorbic acid fatty esters produced by this procedure exhibited a significant antioxidant activity in a micellar substrate composed of linoleic acid.     

Candida sp. 99-125脂肪酶及其在化学品合成中的应用

传统的酯化或转酯化产品的合成通常需要高温、强酸、强碱等相对苛刻的条件,脂肪酶由于其生物催化过程具有高效、高选择性、条件温和和环境友好等特点,在化学品的合成中越来越受到人们的关注。本课题组开发了一种可以用于酯类合成的新脂肪酶,并且实现了该酶的工业化生产。来源于Candida sp. 99-125的脂肪酶在非水相中对酯化和转酯化反应具有高效的催化活性和稳定性。本文介绍了该脂肪酶的发酵生产及其在中长链脂肪酸酯、二元酸酯、维生素A棕榈酸酯、手性化合物以及生物柴油等多种化学品的合成中的应用。