无溶剂体系脂肪酶催化制阿魏酸双油酸甘油酯

在无溶剂体系中以分散在硅藻土上的褶皱假丝酵母脂肪酶为催化剂,以阿魏酸乙酯与油酸甘油酯为原料,一步合成阿魏酸双油酸甘油酯。通过质谱和红外光谱对薄层层析分离纯化后的产物结构进行了表征。定量分析表明,当反应温度为50℃,n(阿魏酸乙酯)/n(油酸甘油酯)=1时,阿魏酸双油酸甘油酯的质量占生成物总质量分数较高。在1 mmoL阿魏酸乙酯,1 mmoL油酸甘油酯和70 mg脂肪酶组成的无溶剂反应体系中,50℃空气浴振荡120 h,总产率可达63.32%,其中阿魏酸双油酸甘油酯质量占生成物总质量的63.37%。     

非水相脂肪酶催化阿魏酸淀粉酯合成

以玉米淀粉和阿魏酸乙酯为原料,利用脂肪酶催化合成阿魏酸淀粉酯。对反应介质和脂肪酶进行了筛选,同时对影响合成阿魏酸淀粉酯反应的因素进行了探究,主要考察了底物摩尔比、酶添加量、反应时间及反应温度等参数对该反应的影响。采用紫外分光光度计对取代度进行测定,并以取代度为考察指标,确定了最佳反应条件：Novozym435脂肪酶为催化剂、异辛烷为反应介质、底物摩尔比为3:1、酶添加量为10%、反应时间18 h、反应温度65 ℃,在此条件下,产物的最大取代度可达0.031. 并通过FT-IR以及1H NMR对产物进行表征。     

非水相脂肪酶催化阿魏酸淀粉酯的合成

以玉米淀粉和阿魏酸乙酯为原料,利用脂肪酶催化合成了阿魏酸淀粉酯,对反应介质和脂肪酶进行了筛选,同时对影响合成阿魏酸淀粉酯反应的因素进行了探究,主要考察了底物摩尔比、酶添加量、反应时间及反应温度对反应的影响。采用紫外分光光度计对取代度进行测定,并以取代度为考察指标,确定了最佳反应条件:Novozym435脂肪酶为催化剂、异辛烷为反应介质、底物摩尔比n(阿魏酸乙酯)∶n(活化淀粉)=3∶1、酶添加量为10%(以活化淀粉质量计)、反应时间18 h、反应温度65℃,在该条件下,产物的最大取代度可达0.031,并通过FTIR以及1HNMR对产物结构进行了表征。     

非水相酶促不可逆转酯反应合成阿魏酸三油酸甘油酯

在有机相甲苯中通过阿魏酸乙烯酯(VF)和三油酸甘油酯(TO)的酶促转酯反应合成油脂抗氧化剂阿魏酸三油酸甘油酯。用阿魏酸乙烯酯作为底物可使转酯反应不可逆,这有利于产物产率的提高并缩短反应时间。通过核磁共振波谱和质谱对阿魏酸乙烯酯的结构进行了表征,并考察了底物摩尔比、反应时间、反应温度、酶用量、水活度对产物产率的影响。结果表明,以甲苯为溶剂,当底物n(VF)∶n(TO)=1∶3,反应时间62 h,反应温度55℃、酶用量20 g/L,水活度(aw)为0.07时,产物的产率最大,达96.73%。     

酶催化茶油转酯化反应制备生物柴油的研究

研究了茶油在脂肪酶Novozym435催化下的转酯化反应,考察了几种直链和支链醇对转酯化反应的影响.选择甲醇作为酰基受体,最佳用量为醇油摩尔比3∶1.通过在反应体系中加入硅胶的策略来提高转酯化反应速率和最终转化率,当硅胶用量为0.8 g/g,30 h的转化率从原来的40%提高到90%.在最佳条件下进行批式反应,酶重复使用15批次后活性无明显下降.     

固定化脂肪酶催化合成生物基增塑剂2,5-呋喃二甲酸正丁酯

以固定化脂肪酶Novozym435为催化剂,在甲苯中催化2,5-呋喃二甲酸二甲酯与正丁醇转酯化反应制备2,5-呋喃二甲酸正丁酯(DBF),对酶促合成DBF的工艺进行优化,探讨了糖类物质和吸附剂作为添加剂对DBF收率的影响。结果表明,在n(2,5-呋喃二甲酸二甲酯)∶n(正丁醇)=1∶5,其中,2,5-呋喃二甲酸二甲酯的物质的量为1 mmol,固定化脂肪酶Novozym435添加量为925 U的10 mL甲苯中,45℃、150 r/min反应28 h后,DBF的收率达到最大。4A分子筛的加入能明显促进DBF的生成,海藻糖的加入也增强了反应体系中脂肪酶的催化能力,在反应体系中加入0.2 kg/L 4A分子筛和固定化脂肪酶质量15%的海藻糖,45℃反应14 h,DBF的收率达到了87.25%,固定化脂肪酶Novozym435在DBF的最优合成体系中连续使用10次后,DBF的收率仍然能保持在47.68%。     

叔戊醇体系酶促大豆油制备生物柴油

叔戊醇作为反应介质,固定化脂肪酶Novozym 435催化大豆油与甲醇的转酯反应制备生物柴油。叔戊醇消除了反应底物甲醇及反应副产物甘油对酶活的负面影响。定量分析表明,叔戊醇与油脂的体积比为1,甲醇与油脂的摩尔比为3,2%脂肪酶,反应体系含水量2%,40 ℃、180 r/min条件下反应15 h,生物柴油得率可达97%。在最适条件下反应进行160批次,酶仍保持了较高的活性和良好的稳定性。     

非水相酶促合成萘普生油酸甘油酯前药

在非相有机试剂异辛烷中通过固定化脂肪酶Novozym435催化萘普生甲酯与三油酸甘油酯反应,通过薄层层析色谱与高效液相色谱对反应液进行分析,并考察了底物摩尔比、反应时间、反应温度、酶添加量、溶剂添加量对转化率的影响。结果表明,当三油酸甘油酯与萘普生甲酯摩尔比3.85,反应时间144h,反应温度50℃,酶添加量150mg,溶剂添加量5m L时,转化率最高达到65.91%。     

非水相酶促合成萘普生油酸甘油酯前药

在非相有机试剂异辛烷中通过固定化脂肪酶Novozym435催化萘普生甲酯与三油酸甘油酯反应,通过薄层层析色谱与高效液相色谱对反应液进行分析,并考察了底物摩尔比、反应时间、反应温度、酶添加量、溶剂添加量对转化率的影响。结果表明,当三油酸甘油酯与萘普生甲酯摩尔比3.85,反应时间144h,反应温度50℃,酶添加量150mg,溶剂添加量5m L时,转化率最高达到65.91%。     

无溶剂体系中癸酸甘油酯的酶法合成

本文通过探索无溶剂体系中固定化脂肪酶催化甘油和癸酸酯化合成癸酸甘油酯的反应特性。对无溶剂体系中酶法催化合成中链甘油酯的可行性进行了研究。     

Enzymatic Glycerolysis of Palm and Palm Kernel Oils

Glycerolysis of palm and palm kernel oils were carried out using commercial lipases from Candida antarctica (Novozym 435) and Mucor miehei (Novozym 388) as catalyst (500 units lipase/g oil) at 40°C and with an oil:glycerol molar ratio of 1:2 in a solvent-free system. Novozym 435 catalyzed the glycerolysis of palm and palm kernel oils giving reaction products in similar compositions. Partial acylglycerols contents of the glycerolysis products obtained from palm and palm kernel oils were 64% (wt) and 66% (wt), respectively. However, partial acylglycerols contents of the glycerolysis products obtained from palm and palm kernel oils conducted with Novozym 388 as catalyst at the same conditions were 44% (wt) and 56% (wt), respectively. On the other hand, free fatty acid contents of the glycerolysis products of palm and palm kernel oils obtained using Novozym 388 were higher, 25–30% (wt), than those obtained by Novozym 435, 4–5% (wt). The monoacylglycerols fraction with the highest content of oleic acid, 62.7% (wt), was obtained from the palm kernel oil glycerolysis reaction catalyzed by Novozym 435.     

Enzymatic Glycerolysis of Palm and Palm Kernel Oils

Glycerolysis of palm and palm kernel oils were carried out using commercial lipases from Candida antarctica (Novozym 435) and Mucor miehei (Novozym 388) as catalyst (500 units lipase/g oil) at 40°C and with an oil:glycerol molar ratio of 1:2 in a solvent-free system. Novozym 435 catalyzed the glycerolysis of palm and palm kernel oils giving reaction products in similar compositions. Partial acylglycerols contents of the glycerolysis products obtained from palm and palm kernel oils were 64% (wt) and 66% (wt), respectively. However, partial acylglycerols contents of the glycerolysis products obtained from palm and palm kernel oils conducted with Novozym 388 as catalyst at the same conditions were 44% (wt) and 56% (wt), respectively. On the other hand, free fatty acid contents of the glycerolysis products of palm and palm kernel oils obtained using Novozym 388 were higher, 25-30% (wt), than those obtained by Novozym 435, 4-5% (wt). The monoacylglycerols fraction with the highest content of oleic acid, 62.7% (wt), was obtained from the palm kernel oil glycerolysis reaction catalyzed by Novozym 435.     

Enzymatic production of alkyl esters through alcoholysis: A critical evaluation of lipases and alcohols

This paper focuses on a detailed evaluation of commercially available immobilized lipases and simple monohydric alcohols for the production of alkyl esters from sunflower oil by enzymatic alcoholysis. Six lipases were tested with seven alcohols, including straight and branched-chain primary and secondary alcohols. The reactions were conducted in a batch stirred reaction vessel using stoichiometric amounts of substrates under solvent-free conditions. Dramatic differences in alcoholysis performance were observed among the different lipases. For most of the alcohols, Novozym 435 produced the highest yield of FA alkyl esters, with yields well over 90% for methanol, absolute ethanol, and 1-propanol. Overall, 96% ethanol was the preferred alcohol for all lipases except Novozym 435, and ethanolysis reactions reached the maximal conversion efficiency. Increasing the water content in the system resulted in an increased degree of conversion for all lipases except Novozym 435. The secondary alcohol 2-propanol significantly reduced the alcoholysis reaction with all lipases; however, the branch-chain isobutanol was more advantageous than linear 1-butanol for Novozym 435, Lipozyme RMIM, and Lipase PS-C. Many commercial immobilized lipases are highly efficient and promising for the production of alkyl esters, offering high reaction yields and a simple operation process.     

无溶剂体系中酶催化合成共轭亚油酸甘油酯

采用直接酯化法,以共轭亚油酸(简称CLA)和甘油为原料,用脂肪酶Novozym 435催化合成共轭亚油酸甘油酯。结果表明:n(甘油)/n(CLA)=5,酶添加量为体系总质量的4%,65℃反应6 h后,共轭亚油酸的转化率为98.18%。所得共轭亚油酸甘油酯为淡黄色油状透明液体,酸价低于3,Novozym 435的操作半衰期为30 h。     

Efficient esterification of sorbitan oleate by lipase in a solvent-free system

The lipase-catalyzed esterification of sorbitan with oleic acid in a solvent-free system to form sorbitan oleate (commercial name Span80) was studied as a feasible approach aimed at meeting the demand for sugar alcohol-based surfactants. Screened results obtained from enzymatic synthesis of sorbitan oleate indicated that Novozym 435 had its highest catalytic activity in a solvent-free system. The introduction of a reduced-pressure system increased the production of sorbitan oleate to a maximum of 95% of theoretical, obtained from 0.2 mol sorbitan, 0.1 mol oleic acid, and 2.0 g lipase (6 wt% of sorbitan) in a solvent-free reaction mixture at optimal reaction conditions. Results obtained from lipase-catalyzed batch esterification reactions showed that more than 90% conversion of sorbitan oleate was maintained after 10 batches of esterification reactions, indicating excellent enzyme stability. Subsequent analysis by HPLC indicated that the product of enzyme-catalyzed esterification by the immobilized lipase contained a significantly greater amount of monoester (about 80%) compared to the composition obtained by chemical synthesis (about 50%).     

Synthesis and component confirmation of biodiesel from palm oil and dimethyl carbonate catalyzed by immobilized-lipase in solvent-free system

The transesterification of palm oil and dimethyl carbonate (DMC) for preparing biodiesel has been carried out at the catalysis of immobilized-lipase in solvent-free system. The components were all confirmed by GC and GC–MS analysis. The fatty acid methyl esters (FAMEs) were analyzed with internal standard method. The fatty acids glycerol carbonate esters (FAGCs) were characterized as the intermediates. And, glycerol dicarbonate (GDC) was confirmed as the byproduct by comparing with the model compound. Moreover, the effects of the reaction conditions (type of lipases, molar ratio of DMC and palm oil, amount of catalyst, reaction temperature and time) on the yield of FAMEs were investigated. The yield of FAMEs could reach 90.5% at 55 °C for 24 h with the molar ratio of DMC to oil 10:1 and the catalyst amount of 20% Novozym 435 (based on the oil weight). There was no obvious loss in the FAMEs yield after Novozym 435 having been used for eight cycles.     

Biosysthesis of corn starch palmitate by lipase novozym 435

Esterification of starch was carried out to expand the usefulness of starch for a myriad of industrial applications. Lipase B from Candida antarctica, immobilized on macroporous acrylic resin (Novozym 435), was used for starch esterification in two reaction systems: micro-solvent system and solvent-free system. The esterification of corn starch with palmitic acid in the solvent-free system and micro-solvent system gave a degree of substitution (DS) of 1.04 and 0.0072 respectively. Esterification of corn starch with palmitic acid was confirmed by UV spectroscopy and IR spectroscopy. The results of emulsifying property analysis showed that the starch palmitate with higher DS contributes to the higher emulsifying property (67.6%) and emulsion stability (79.6%) than the native starch (5.3% and 3.9%). Modified starch obtained by esterification that possesses emulsifying properties and has long chain fatty acids, like palmitic acid, has been widely used in the food, pharmaceutical and biomedical applications industries.     

Lipase-catalyzed acidolysis of menhaden oil with pinolenic acid

Lipase-catalyzed acidolysis of menhaden oil with a pinolenic acid (PLA) concentrate, prepared from pine nut oil, was studied in a solvent-free system. The PLA concentrate was prepared by urea complexation of the FA obtained by saponification of pine nut oil. Eight commercial lipases from different sources were screened for their ability to catalyze the acidolysis reaction. Two different types of structured lipids (SL) were synthesized. The first type, which has PLA residues as a primary FA residue at the sn-1,3 positions of the TAG, was synthesized using a 1,3-regiospecific lipase, namely, Lipozyme RM IM from Rhizomucor miehei. The second type of SL, which has PLA residues as a primary FA residue at both the sn-1,3 and sn-2 positions of the TAG, was synthesized using a nonspecific lipase, namely, Novozym 435 from Candida antarctica. The effects of variations in enzyme loading, temperature, and reaction time on PLA incorporation into the oil were monitored by GC analyses. The optimal temperature and enzyme loading for synthesis of the two types of SL were 50°C and 10% of the total weight of substrates for both enzymes. The optimal reaction time for the synthesis with Lipozyme RM IM was 16h, whereas the optimal reaction time for the synthesis mediated by Novozym 435 was 36 h. Pancreatic lipase-catalyzed sn-2 positional analyses were also carried out on the TAG samples.     

固定化脂肪酶合成肉豆蔻酸异丙酯的研究

采用固定化脂肪酶(Novozym 435)在有机溶剂体系中催化肉豆蔻酸和异丙醇酯化合成了肉豆蔻酸异丙酯.研究了各种因素如有机溶剂种类、反应物摩尔比、反应温度及酶用量等因素对酯化率的影响.结果表明,肉豆蔻酸异丙酯合成的较佳条件为:1.14 g肉豆蔻酸,0.45 g异丙醇,0.02 g Novozym 435,10 mL石油醚(其中加入0.5 g 3A分子筛吸水),在55℃下振荡反应4 h,酯化率为99.2%(肉豆蔻酸可基本完全转化为肉豆蔻酸异丙酯).实验过程中固定化酶稳定性较好,可连续使用7批左右.     

Optimization of the synthesis of lower glycerides rich in unsaturated fatty acid residues obtained via enzymatic ethanolysis of sesame oil

The lipases Novozym 435, Lipozyme TL IM and Lipozyme RM IM were employed in the production of lower acylglycerols (LG), i.e. mono‐ (MAG) and diacylglycerols (DAG), rich in unsaturated fatty acids from sesame oil in batch reactors. The effect of the molar ratio of ethanol to fatty acids on the reusability of these immobilized lipases was studied in detail. The effects of pretreatment on lipase activity for ethanolysis were investigated. Glycerol had a strong product inhibition effect on the ethanolysis reaction, and a relatively large excess of ethanol was necessary to remove the glycerol adsorbed on these biocatalysts. The enzymatic activity was drastically reduced by addition of water to the reaction medium. The presence of organic solvents (hexane and acetone) did not favor the production of LG. For the Novozym 435‐catalyzed reaction, optimum conditions were a molar ratio of ethanol to fatty acid residues of 5 : 1, 15 wt‐% lipase and 50 °C. For Lipozyme TL IM, the optimum conditions were a molar ratio of ethanol to fatty acid residues of 5 : 1, 20 wt‐% biocatalyst, and 30 °C. Novozym 435 and Lipozyme TL IM produced LG with molar ratios of unsaturated to saturated fatty acids of 20.4 in 1 h and 25.3 in 5 h, respectively. In the original oil, this ratio was 5. For trials conducted under optimum conditions, the products from the Novozym 435 trials contained 21.8 wt‐% triacylglycerols (TAG), 24 wt‐% DAG and 54.2 wt‐% MAG. The products of the Lipozyme TL IM trials consisted of 12.9 wt‐% DAG and 87.1 wt‐% MAG. No TAG species were detected.