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研究了水/有机溶剂两相体系中,出芽短梗霉SW0202细胞催化4-氯乙酰乙酸乙酯(COBE)不对称还原制备(S)-4-氯-3-羟基丁酸乙酯(CHBE)的过程.有机溶剂的选择表明邻苯二甲酸二丁酯为该反应最适有机溶剂,与单一水相体系相比,水/邻苯二甲酸二丁酯两相体系能明显改善反应效率.通过系统考察一些因素如相体积比、振摇速度、反应温度和pH等对反应速度、摩尔转化率和产物光学纯度的影响发现,这些因素对还原反应的初速度和摩尔转化率有较明显的影响,而对产物的光学纯度影响不明显.优化后的条件为:水与有机溶剂的相体积比1∶1,振摇速度180 r·min-1,温度30℃,pH 7.0.在该优化反应条件下,水/有机溶剂两相体系中出芽短梗霉SW0202细胞催化COBE生成(S)-CHBE不对称还原反应的最大摩尔转化率和产物的光学纯度分别达到了94.8%和97.9%e.e.,在用Na2CO3溶液控制水相pH 7.0条件下,有机相中的产物浓度积累达58.2 g·L-1. 相似文献
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以DL-苯丙氨酸为底物,利用巨大芽孢杆菌(Bacillus megaterium)AS1.127苯丙氨酸脱氨酶光学异构选择性,仅对L-苯丙氨酸专一氧化脱氨,而不作用于D型,从而高效制备D-苯丙氨酸。考察了影响酶促反应的几种因素,得到了最佳酶促反应条件,结果表明,该转化反应最适温度37~40℃。最适pH 5.8,0.2%磷酸缓冲液可提高脱氨酶活力,在反应液中加入表面活性剂0.2%吐温-80或10-5mol/L K 能显著提高脱氨酶活力。转化率与底物浓度、菌体用量有关,在此反应体系中,底物DL-苯丙氨酸浓度0.09 mol/L,菌体用量0.03 g/mL,转化时间18 h,L-苯丙氨酸转化率最高达98%以上。转化液经脱色、减压浓缩和等电点结晶后得D-苯丙氨酸,比旋光为[a]2D0= 32.4°,光学纯度达到95%以上。 相似文献
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含离子液体介质中脂肪酶高效催化对羟基苯甘氨酸甲酯不对称氨解 总被引:1,自引:1,他引:0
利用离子液体C6MIm·BF4 (20%,v/v)/叔丁醇混合介质中脂肪酶Novozym 435催化对羟基苯甘氨酸甲酯不对称氨解反应,高效地制备了对映体纯D-对羟基苯甘氨酸酰胺.该反应的最适氨源为氨基甲酸铵,其较佳浓度为120~160 mmol·L-1;最适水活度、反应温度、pH值、对羟基苯甘氨酸甲酯浓度分别为0.55~0.75、35~40℃、7.5~8.0和40 mmol·L-1.在上述条件下,反应初速度和对映体选择比分别为5.88 mmol·L-1·h-1和83 (E值),反应6 h,产率为47.6%,产物e.e.值为92.9%.与有机溶剂叔丁醇相比,脂肪酶Novozym 435在C6MIm·BF4 (20%,v/v)/叔丁醇混合溶剂中的操作稳定性显著提高,反应7批次后,仍能维持90%以上的催化活性,而在叔丁醇反应介质中,仅能维持41%的催化活性. 相似文献
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提出以病死畜禽为原料制备氨基酸镁新工艺,主要过程是先将病死畜禽尸体水解为富含大量氨基酸的营养液,并进一步加入无机镁盐使Mg2+与氨基酸螯合成氨基酸镁。通过单因素实验,确定了病死鸡的最佳催化水解条件,并进一步探究了水解氨基酸制备氨基酸镁的反应条件。水解反应的最佳条件是:甲酸加入量10 mL,水加入量75 mL,病死鸡尸体组织加入量50 g,反应温度为96℃,水解时间为5 h,加料方式为多次加料。螯合反应的最佳条件是:pH=3.0,反应时间为1 h,反应温度为50℃,n(氨基酸)∶n(氯化镁)=1∶1,在最佳反应条件下,氨基酸镁螯合率可达到93.49%。 相似文献
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两相体系生物转化L-苯丙氨酸生成2-苯乙醇 总被引:7,自引:0,他引:7
研究了水/有机溶剂两相体系中,酵母细胞转化L-苯丙氨酸(L-phe)生成2-苯乙醇(PEA)的过程.菌种选择表明酿酒酵母CICIM Y0086(T)的转化能力要高于马克斯克鲁维酵母AS.2.1440,有机溶剂的选择表明,在实验使用的4种有机溶剂中,油酸为该反应最适有机溶剂.当初始底物浓度为6 g/L、水和油酸的相体积比为1:1、反应温度30 ℃和pH值4.0、装液量20 mL/250 mL 的条件下,转化48 h,产物浓度达到3.0 g/L,比单水相提高了50%. 相似文献
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为了高效地合成高光学纯手性亚砜,在单水相体系中利用Rhodococcus sp. CCZU10-1选择性氧化拆分外消旋苯甲亚砜(rac-PMSO)合成了(S)-PMSO(ee>99.9%)。通过考察反应pH值、反应温度、摇床转速、辅助底物、生物催化剂添加量对催化反应的影响,确定了最适反应条件。结果表明最适反应条件为:反应pH值8.0、反应温度30℃、摇床转速180r/min、辅助底物为半乳糖(50mmol/L)、细胞浓度为0.08g(湿重)/mL。在最适反应条件下生物转化20mmol/L rac-PMSO时,(R)-PMSO完全转化,(S)-PMSO(ee>99.9%)产率为47.1%。因此,研究结果为工业化生产(S)-PMSO奠定了基础。 相似文献
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氨基树脂固定胃蛋白酶的方法及性质研究 总被引:1,自引:0,他引:1
采用氨基树脂作为载体,戊二醛作为交联剂,对胃蛋白酶的固定化进行了研究,并对固定化条件和固定化胃蛋白酶的部分酶学性质进行了分析。确定固定条件为:戊二醛浓度为5%,载体处理温度为室温(25℃),处理时间为5h,m(胃蛋白酶):m(氨基树脂)为1:25,pH为3.0,固定时间为12h。此条件下固定化的胃蛋白酶活力为30U/g,酶的活力回收率为60%。与非固定化相比最适水解温度由50℃升高到60℃,最适pH值由2.0升高到4.0,游离酶米氏常数3.08g/L,固定化酶米氏常数1.2g/L,固定化胃蛋白酶的储存半衰期约为25天。对珠蛋白的操作半衰期为9天。 相似文献
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分别采用乳化交联法和共沉淀法制备磁性壳聚糖微球载体,并对形貌结构进行比较,结果表明,采用共沉淀法制备的磁性壳聚糖微球负载Fe3O4的效果好,故将其作为载体固定甲酸脱氢酶。最佳固定化条件:添加酶量9 U.g-1,pH=7.0,固定化时间5 h。游离酶和固定化酶的最适宜反应温度分别为50℃和30℃;游离酶的最适宜pH=7.0,固定化酶的最适宜pH=6.0;将游离酶和固定化酶分别置于60℃恒温水浴放置180 min后,游离酶和固定化酶的相对酶活力分别为0.78%和40.39%;将游离酶和固定化酶置于不同pH的缓冲液中保存1 h后,在强酸(pH=2.0)和强碱(pH=10.0)条件下,固定化酶的相对酶活力分别为11.03%和38.43%,游离酶已全部失活;固定化酶重复使用6次后,相对酶活力为73.53%,表明固定化酶具有较好的热稳定性、酸碱稳定性和操作稳定性。 相似文献
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苯丙氨酸脱氨酶发酵工艺及其酶学性质研究 总被引:1,自引:0,他引:1
选育出一株具有较高苯丙氨酸脱氨酶活性的菌株巨大芽孢杆菌AS1.127-NJU10。考察了该菌的发酵产酶条件,结果表明:蔗糖为最佳碳源、酵母浸膏和NH4C l组成最佳氮源,其质量浓度分别为:ρ(蔗糖)=20 g/L,ρ(酵母浸膏)=2 g/L和ρ(NH4C l)=10 g/L;发酵培养基最适pH=6.5,培养温度为37℃;诱导物ρ(L-苯丙氨酸)=1 g/L时,酶活最高达1 070 U。同时对苯丙氨酸脱氨酶的性质进行了研究,结果表明:该酶最适pH=5.8,最适温度为40℃,反应液中添加φ(吐温-80)=0.2%和c(K+)=10-5mol/L能明显提高酶活。 相似文献
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Using polyacrylic acid (PAA) modified polyethylene (PE) membrane as a carrier, two immobilization routes of alcohol dehydrogenase (ADH) were studied, and the catalytic performance of immobilized enzyme was investigated using formaldehyde as a substrate. In the first route, PAA-PE membrane was further modified by polyethyleneimine (PEI) and then ADH was covalently linked by glutaraldehyde (GA) to the surface of PEI/PAA-PE. The results show that the optimal immobilization pH was 6.0, immobilization temperature was 5—15℃, ADH and GA concentrations were 1.0mg/ml and 0.01%(mass). For immobilized enzyme, the optimal reaction pH was 6.5, temperature was 15—30℃, and the highest reaction rate was 9.6 μmol/(L·min), the remaining activity was 47.3% after 10 use cycles. In the second route, ADH was immobilized on PAA-PE membrane with 1-(3-dimethylaminopropyl)-2-ethylcarbodiimide hydrochloride (EDC) and N-hydroxysuccinimide (NHS) as activators. The results show that the optimal molar ratio of EDC and NHS was 1∶0.5, and the immobilization time was 24 h. For immobilized enzyme, the optimal reaction pH was 6.5, temperature was 20—37℃, and the highest reaction rate was 15.58 μmol/(L·min), 53.8% activity was remained after 10 cycles. 相似文献
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Maria M Cardoso Mrio J Barradas Karl H Kroner Joo G Crespo 《Journal of chemical technology and biotechnology (Oxford, Oxfordshire : 1986)》1999,74(8):801-811
The aim of this work is to investigate the driving forces involved in amino acid solubilization in cationic reversed micelles, and to determine in which way different parameters affect the reversed micellar structure and amino acid solubilization, in order to select the best conditions to optimize amino acid extraction. To this end, extraction equilibrium experiments were performed using different experimental conditions and three amino acids with different structures: aspartic acid – a hydrophilic amino acid, phenylalanine – a slightly hydrophobic amino acid, and tryptophan – a hydrophobic amino acid. The study of the effect of amino acid related parameters, such as pH and the initial amino acid concentration in the aqueous phase, and the effect of parameters that influence the reversed micellar structure, such as surfactant concentration, ionic strength and co‐surfactant concentration, provides useful information about the driving forces involved, solute–micelle interfacial interactions and solute location in the cationic system trioctylmethylammonium chloride (TOMAC)/hexanol/n‐heptane. These parameters can be adjusted to optimize amino acid extraction. It is shown that amino acids with the same isoelectric point can be selectively separated by exploring the different interactions they establish with the reversed micellar interface. © 1999 Society of Chemical Industry 相似文献
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以聚丙烯酸(PAA)改性的聚乙烯(PE)膜为载体,研究了醇脱氢酶(ADH)的两种固定化路线,并以甲醛为底物考察了固定化酶的催化性能。路线1用聚乙烯亚胺(PEI)进一步改性,使用戊二醛(GA)固定化ADH。最优固定化pH为6.0,温度为5~15℃,酶浓度为1.0 mg/ml,GA浓度为0.01%(质量);固定化酶的最适反应pH为6.5,温度为15~30℃,反应速率最高为9.6μmol/(L·min);重复利用10次后可保持47.3%的活性。路线2以PAA-PE为载体,用1-(3-二甲氨基丙基)-2-乙基碳二亚胺盐酸盐(EDC)和N-羟基琥珀酰亚胺(NHS)为活化剂,固定化ADH。EDC和NHS最优摩尔比为1∶0.5,固定化时间为24 h;固定化酶的最适反应pH为6.5,温度为20~37℃,反应速率为15.58μmol/(L·min);重复利用10次后可保持53.8%的活性。 相似文献
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R. S. Farag S. A. Osman S. A. S. Hallabo A. A. Nasr 《European Journal of Lipid Science and Technology》1981,83(2):62-65
The oxidation of refined cotton seed oil catalysed by various α-amino acids and albumin have been studied in aqueous and non-aqueous media. Cysteine, phenylalanine and albumin possessed pro-oxidant effect in cotton seed oil in aqueous and non-aqueous media. Serine exhibited pro-oxidant activity in aqueous media and minor anti-oxidant effect in non-aqueous media. The effectiveness of the amino acids on cotton seed oil oxidation was in the following descending order in both aqueous and non-aqueous media: cysteine > phenylalanine > serine. The pro-oxidant effect in aqueous media might be due to the predominant presence of the protonated amino nitrogen. Whilst, amino acid-metal complex might be responsible for the pro-oxidant effect in non-aqueous media. 相似文献
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Hong-Sik Hwang Jill K. Winkler-Moser Sean X. Liu 《Journal of the American Oil Chemists' Society》2022,99(5):407-419
Previous studies reported that several amino acids had strong antioxidant activity in vegetable oils under frying conditions. In this study, amino acids were converted to their sodium or potassium salts, and a heating study was conducted with 5.5 mM amino acid salts in soybean oil (SBO) at 180°C. Sodium salts of amino acids including alanine, phenylalanine, and proline and disodium glutamate had significantly stronger antioxidant activity than the corresponding amino acids, and potassium salts had stronger antioxidant activity than sodium salts. Potassium salts of alanine and phenylalanine more effectively retained tocopherols in SBO than the corresponding amino acids during heating. Phenylalanine potassium salt had stronger antioxidant activity than phenylalanine in other vegetable oils including olive, high oleic soybean, canola, avocado, and corn oils. Phenylalanine potassium salt at 5.5 mM more effectively prevented oil oxidation than tert-butyl hydroquinone, a synthetic antioxidant, at its legal concentration limit (0.02%) indicating its feasibility as a new antioxidant for frying. 相似文献
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Danae Doulia Fotis Rigas C Gimouhopoulos 《Journal of chemical technology and biotechnology (Oxford, Oxfordshire : 1986)》2001,76(1):83-89
The adsorption of eight amino acids, L ‐asparagine, D,L ‐threonine, L ‐lysine, L ‐leucine D,L ‐methionine, L ‐tyrosine, L ‐phenylalanine and D,L ‐tryptophan, on the non‐polar macroporous adsorbents Amberlite XAD‐2 and XAD‐4 (polystyrene–divinylbenzene copolymers) was studied. Equilibrium adsorption experiments were conducted to estimate the types of isotherm and their parameters. The effect the chemical composition and structure of the amino acids on the efficiency of adsorption was evaluated. The influence of pH and ionic strength was also studied. The data of adsorption isotherms of the examined amino acids seemed generally to approach the Freundlich isotherm model. Tryptophan isotherm adsorption data could match in some cases the Langmuir model. The majority of the adsorption isotherms were almost linear. In terms of adsorbed amino acid on both resin surfaces, the amino acids can be ranked thus: D,L ‐tryptophan > L ‐phenylalanine > D,L ‐methionine, L ‐tyrosine > L ‐leucine > L ‐lysine > D,L ‐threonine > L ‐asparagine. In low pH solution, adsorption was generally higher than that at intermediate and high pH values. Generally, as the ionic strength increases, the adsorption of the amino acids increases. © 2001 Society of Chemical Industry 相似文献