产脂肪酶海洋酵母Bohaisea-9145批式发酵动力学研究

以从渤海海域海泥中分离获得一株产脂肪酶海洋酵母Bohaisea-9145为实验菌株,为认识其发酵机理并指导发酵工艺设计,利用批式发酵方法研究该酵母菌体生长和菌体产酶情况。该酵母菌发酵类型为生长非关联型,发酵溶氧浓度最佳为30%±2%,当限制性基质葡萄糖浓度达到2.2772g/L时细胞生长速率最大,通过数学推导和批式发酵结果建立了海洋酵母菌体生长和产脂肪酶动力学模型。      

反胶束体系中脂肪酶催化乙酸甲酯与棉籽油合成生物柴油

在丁二酸二酯磺酸钠(AOT)反胶束体系中,采用游离脂肪酶催化棉籽油与乙酸甲酯反应制备生物柴油。考察了AOT浓度、水与AOT的质量比(ζ0)、脂肪酶用量、缓冲溶液pH、乙酸甲酯与棉籽油的摩尔比(r)、反应温度、摇床转速等因素对脂肪酶催化合成生物柴油的影响,并考察了脂肪酶的重复使用性能。经单因素实验及正交实验,得出优化的反应条件:AOT浓度40mmol/L,ζ0=7,脂肪酶用量10mg(溶液体积为2.5mL),缓冲溶液pH=7,r=3,反应温度35℃,摇床转速160r/min。在优化的反应条件下,产物脂肪酸甲酯的单程收率最高可达65.53%。酯交换制备生物柴油的影响因素大小依次为:缓冲溶液pH>反应温度>r>脂肪酶用量>AOT浓度>摇床转速>ζ0。与甲醇和碳酸二甲酯作为酰基受体相比,乙酸甲酯为酰基受体时脂肪酶的稳定性较好。     

微乳凝胶固定化脂肪酶及其催化酯化反应的研究

描述了以ＡＯＴ／异辛烷／水／明胶组成的反相微乳凝胶的制作过程,作出了不同ＡＯＴ浓度下的凝胶相图。以此反相微乳凝胶固定脂肪酶,在有机溶剂中催化脂肪酸 和脂肪醇酯化反应,转化率可达９５％以上,在０℃时有有较快的反应速度,固定化酶重复使用２９次,反应初速度仍有７０％。     

基于BP神经网络和GA研究啤酒糟不溶性膳食纤维的酶法脱脂工艺

采用脂肪酶对酶碱法制备啤酒糟不溶性膳食纤维(IDF)的脱脂工艺进行研究,并对制备得到的IDF 成分和功能特性进行分析。在正交试验的基础上,基于BP 神经网络建立脂肪酶脱脂模型,利用遗传算法优化工艺条件。BP 神经网络建立的脱脂模型误差为0.0001,具有较强的逼近能力。优化得到的最佳工艺条件是加酶量0.7g、酶解温度39.6℃、酶解时间5.6h。在此条件下,脂肪去除率达74.1%。制备得到IDF 的溶胀性达6.05mL/g,持水力达318.2%,具有较好的生理活性。     

不同脂肪酶催化亚麻油水解反应性能的比较

从反应温度、水／亚麻油摩尔比、脂肪酶用量等方面比较了L-lipase和N—1ipase脂肪酶在无溶剂体系中对亚麻油水解反应的催化性能。得到了两种脂肪酶的最佳使用温度及催化亚麻油水解反应的适宜条件。结果表明，两种脂肪酶的最佳用量均为3％～5％，适宜的水／亚麻油摩尔比为30：1。L-lipase和N-lipase的最佳温度分别是35℃和67．5℃，与L-lipase相比，N—1ipase的热稳定性和催化活性相对较高。     

Enzymatic Synthesis of Glucose- and Xylose Laurate Esters Using Different Acyl Donors,Higher Substrate Concentrations,and Membrane Assisted Solvent Recovery

Glucose- and xylose laurate esters are enzymatically synthesized using equimolar substrate concentrations in 2-methyl-2-butanol, comparing free lauric acid with methyl- and vinyl-laurate as acyl donors. All reactions result in ≥70% acyl donor conversions after 72 h but the activated donors are also partially hydrolyzed to lauric acid, highlighting the difficulty in controlling water presence in this particular reaction system. The esterification of xylose generates a complex product profile, with several regioisomers of monoesters and diesters. The esterification of glucose is quite selective, forming mainly the 6-O monoester (≥96%) with a small presence of two diester isomers (4%). Increasing substrate concentration up to 800 millimoles kg⁻¹ results in lower conversion values (down to 58%) but shows that the reaction proceeds successfully even in the presence of high amounts of insoluble glucose. However, the reaction is less selective and the proportion of diester increases, becoming up to 46% (molar fraction) of the final product. Solvent recovery after esterification can be achieved by organic solvent nanofiltration through a polymeric membrane able to retain ≥80% of all reaction substrates and products. Practical Applications: The use of high substrate concentrations during the enzymatic synthesis of sugar ester biosurfactants leads to product titers that are more industrially appealing, without the need to find a solvent that can solubilize all initial substrate. The sustainability of the enzymatic conversion at mild temperatures can be enhanced by recycling of the reaction solvent through organic solvent nanofiltration, an energy efficient alternative to other traditional methods like distillation.     

Chiral resolution of differently substituted racemic acetyl‐1‐phenyl ethanol using lipase from Bacillus subtilis

In the present investigation a Lipase producing strain, Bacillus subtilis (MTCC‐121) was grown on various media containing different sources of carbon, nitrogen and other nutrients. The best media found for the production of lipase was M2 media containing 0.4% peptone, 0.2% beef extract and 1% NaCl. Lipase produced from this culture was used for the kinetic resolution of racemic acetyl‐1‐phenyl ethanol and its derivatives, which are important as chiral auxiliaries and intermediates in the synthesis of natural products, pharmaceuticals and agrochemicals. The lipase resolved these substrates after 48 h with enantiomeric excess of 90–98% and conversion 40–48%. Copyright © 2010 Society of Chemical Industry     

A continuous ultrasound‐assisted packed‐bed bioreactor for the lipase‐catalyzed synthesis of caffeic acid phenethyl ester

BACKGROUND: The focus of this paper is the ultrasound‐assisted synthesis of caffeic acid phenethyl ester (CAPE) from caffeic acid and phenyl ethanol in a continuous packed‐bed bioreactor. Immobilized Novozym^® 435 (from Candida antarctica) is used as the catalyst. A three‐level–three‐factor Box–Behnken design and a response surface methodology (RSM) are employed to evaluate the effects of temperature, flow rate, and ultrasonic power on the percentage molar conversion of CAPE. RESULTS: Based on ridge max analysis, it is concluded that the optimum condition for synthesis is reaction temperature 72.66 °C, flow rate 0.046 mL min^?1, and ultrasonic power 1.64 W cm^?2. The expected molar conversion value is 97.84%. An experiment performed under these optimal conditions resulted in a molar conversion of 92.11 ± 0.75%. The enzyme in the bioreactor was found to be stable for at least 6 days. CONCLUSIONS: The lipase‐catalyzed synthesis of CAPE by an ultrasound‐assisted packed‐bed bioreactor uses mild reaction conditions. Enzymatic synthesis of CAPE is suitable for use in the nutraceutical and food production industries. Copyright © 2011 Society of Chemical Industry     

双氧水对米赫根毛霉脂肪酶在米曲霉中重组表达的调控研究

米赫根毛霉脂肪酶RML是具有广泛应用价值的重要微生物脂肪酶。本研究对双氧水调控米曲霉RML转化子ONL1表达脂肪酶进行了研究。荧光定量PCR和SDS-PAGE表明,在转化子培养过程中,维持培养体系中10 mmol/L双氧水2 h,使转化子ONL1的脂肪酶活力提高5倍。在双氧水处理过程中,RML的翻译未受影响,其表达水平的提高源于双氧水对其转录水平的调控。因此,双氧水调控melO启动子控制的外源基因的表达体现在转录水平。由于双氧水易挥发,导致其效果低于延长培养时间。为了在较短培养时间内获得高酶活,应在培养液中持续添加双氧水（10 mmol/L）。基于米曲霉转化子脂肪酶活力的分析和qPCR检测,确定以下策略可用于提高米曲霉中RML的表达水平：脂肪酶RML基因的密码子优化、信号肽序列优化、连续分批培养。