酶促合成生物柴油反应动力学

以Candidasp.99-125脂肪酶为催化剂,甘油三油酸酯和甲醇为底物,采用有序机制模型对酶促合成生物柴油的酯交换反应动力学进行了研究,并与经典乒乓机制模型进行了比较。研究结果表明,反应初速率的实验值与有序机制模型方程的计算值吻合很好。对于固定化Candidasp.99-125脂肪酶催化合成生物柴油的酯交换反应机理进行研究,采用有序机制模型比经典乒乓机制模型更为精确。反应过程中,醇抑制为竞争性抑制,在甘油三油酸酯浓度较小的范围内,醇抑制作用较为显著,醇浓度越低反应初速率越快。该有序机制模型可用来预测生物柴油的生产批次或连续反应器中酯交换反应的速率,确定最佳底物油脂和醇的浓度。     

粗状假丝酵母(Candida valida T2)生产脂肪酶的发酵条件

对粗状假丝酵母产生脂肪酶的培养条件进行了研究。结果表明,该菌株产脂肪酶的适宜培养基组成为:桐油15mL/L,黄豆粉30g/L,糊精10g/L,硝酸铵10g/L,MgSO4·7H2O1g/L,K2HPO42g/L,Tween800.5g/L;最佳培养为温度30℃、发酵液起始pH6,摇瓶发酵脂肪酶活力达到1467U/mL。     

Combined structural and biological activities for new polyunsaturated fatty derivatives obtained by biotechnological process

The objective of this study is to demonstrate the use of new polyunsaturated fatty derivatives for cutaneous applications. These new compounds present an analogue structure of cutaneous lipid, stabilize the polyunsaturated fatty acids (face to oxidation) and demonstrate specific biological activities. Three molecules described are Omega 6 fatty acid stabilized compound (O6FASC), the O3FASC and the O9FASC. The derivatives are synthesized via the same biotechnological process. This work describes the choice of final structure, the design of the biotechnological process and the free solvent enzymatic synthesis used for the synthesis of these three cutaneous lipid analogues. The restructuring effect of such analogues has been demonstrated with an in vivo study on volunteers. The stabilization of the O3FASC and O6FASC, and the biological activities of these three compounds are presented. The O6FASC shows very good results in anti-inflammatory effects; the O3FASC has anti-stress activities, whereas the O9FASC presents interesting results in improving elasticity and firmness. All these activity tests are presented in this work.     

In vivo lipolysis induced by heparin injection or fasting: a proton MR spectroscopy analysis of human plasma

The impact of induced lipolysis on the composition of plasma lipids is analyzed by proton magnetic resonance spectroscopy (MRS) in humans. The variations of the methylene and methyl resonances from lipids in lipoproteins are studied under two sets of lipolytic conditions: acute endovascular lipolysis induced by an intravenous injection of heparin and subacute lipolysis induced by short fasting. During acute lipolysis, the degradation of the very low density lipoproteins structures is well correlated to the modifications observed in the areas of CH₂ and CH₃ MRS signals. The comparison of regular spectra, spectra with water signal suppression, and spectra recorded with a spin-echo sequence provides information on the behavior of the different parts of the lipoproteins, that is, the neutral core, little affected by heparine-induced lipoprotein lipase (LPL) activation, and the surface layer supplying substrates to LPL. During 48 h of fasting, only limited modifications occur on the MR spectra, and lipolysis cannot be documented in details.Address for correspondence: CRMBM-CNRS, Faculté de Médecine, 27, bd Jean Moulin, 13005 Marseille, France.     

一株产耐高温碱性脂肪酶菌株的筛选及产酶条件优化

用Tween平板法从采集自河北省的122份土样中筛选到一株脂肪酶活性较高的菌株Lipa1318。综合其形态学和生理生化特征以及16S r DNA的序列比对结果,确定该菌株为粘质沙雷氏菌（Serratia marcescens）。其所产胞外脂肪酶的最适反应温度为60℃,最适p H为8.0,是一种耐热的碱性脂肪酶。摇瓶发酵实验证明该菌株的最适产酶条件为:葡萄糖1.0%,牛肉膏1.0%,Triton X-100 1.5%,橄榄油0.50%,Ca Cl20.50mmol/L,发酵液初始p H为8.0,30℃培养96h,通过发酵条件的优化使酶活提高了3.6倍。      

固定化脂肪酶生物反应器催化合成乙酸正己酯

为提高非水相酶促合成食品添加剂乙酸正己酯的效率,以羧甲基纤维素为稳定剂,通过物理吸附,将假单胞菌脂肪酶Pseudomonas cepacia lipase固定在锥形瓶内壁上,制备成简易的生物反应器,并研究了其用于催化合成乙酸正己酯的动力学。结果表明,在反应器中,加入己醇与乙酸乙烯酯的混合液,并在37℃、150r/min下摇动,反应即开始;反应液倒出,则反应停止,无需过滤。反应7h,相对于酶粉,反应器中的固定化酶,可使反应转化率提高7倍,48h后,转化率达99%。经10次循环催化、共10d与有机相的接触,仍保留有52%的转酯活性;同样条件下,不添加羧甲基纤维素的固定化酶,仅有12.8%的活性。可见,这种反应器操作方便,能有效提高酶的非水活性和稳定性,适于催化非水相反应。      

Enhancement of Release of Short-Chain Fatty Acids from Milk Fat with Immobilized Microbial Lipase

Candida cylindracea or Rhizopus arrhizus lipase was entrapped in polymer gels prepared from photo-cross-linkable resin prepolymers. The immobilized enzyme was used in milk fat hydrolysis and effective in enhancing the release of short-chain fatty acids compared with free enzyme. The molar percentage of short-chain fatty acids (butyric acid to capric acid) released in the reaction increased significantly after enzyme immobilization. The gel composed of hydrophobic long-chain prepolymer gave the highest increase, with 5.9-fold and 4.5-fold increases for C. cylindracea and R. arrhizus lipase, respectively. Difference in substrate diffusion rate between milk fat triglycerides may be responsible for this behavior.     

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     

Enzymatic preparation of polyethylene glycol esters of castor oil fatty acids and their surface-active properties

This study concerns the preparation and evaluation of nonionic surfactants prepared from polyethylene glycol (PEG) esters of castor oil fatty acid, a source of hydroxy fatty acid. A lipase-catalyzed esterification reaction has been employed to prepare PEG esters of hydroxy acid to overcome problems associated with chemical processes. Castor oil fatty acid (85% ricinoleic acid) was mixed with PEG of different molecular weight. Rhizomucor miehei lipase was added as catalyst (10% level) and the reaction was continued at 60°C under 2 mm Hg pressure for 360 min. Conversion of PEG to esters was in the range of 86–94%, depending on the molecular size of PEG. The products were isolated and examined for surface activity by surface tension measurement. Surface tension values measured at 25°C were about 36–37 dynes/cm.