Enzymatic alcoholysis reaction of soy phospholipids

Lipase-catalyzed alcoholysis of soy phospholipids was investigated to simultaneously make lysophospholipids and fatty acid esters of individual alcohols. Alcoholysis was carried out by stirring a mixture of soy phospholipids and individual alcohols in equimolar proportions with 10% (by weight of reactants) Mucor miehei lipase at 55°C for 24 h. The products were isolated by column chromatography after removal of the lipase. Lysophospholipids (in 69–78% molar yield) were obtained from soy phospholipids, and the yield of esters of various alcohols also conformed nearly with theoretical yields.     

大豆磷脂酶水解技术研究

为获取制备酶水解磷脂的最佳条件 ,以浓缩磷脂为底物 ,磷脂酶A2 为水解酶 ,采用静止反应方式 ,进行了大豆磷脂酶水解改性研究。最佳反应条件为 :磷脂酶A2 0 .14 % ,氯化钙 0 .0 3% ,时间 7h ,温度 70℃。在最佳条件下 ,FFA的生成量可以达到 2 2 .3mL ,转化率可达到 36 .36 %。FFA的生成量与溶血磷脂的生成量有显著的相关与回归关系 (P <0 .0 1) ,回归方程为 :Y =0 .15 2X +13.6 77。     

Surface activity of the lipid products hydrolyzed with lipase and phospholipase A-2

We examined thein vitro surface activity, immersional wettability and adhesional wettability shown by aqueous solutions of soy lysophospholipid (SLP)/monoglyceride (MG)/fatty acid (FA), SLP/FA and SLP/MG, and found that many lipid mixtures showed significant surface activity when their MG and FA components consisted of polyunsaturated FA and/or medium chain FA. The more unsaturated the FA, the higher the surface activity. A mixture of SLP/medium chain fatty acid MG (medium chain MG)/medium chain FA showed the highest surface activity, and was comparable to an Aerosol-OT surfactant, the most effective wetting agent. SLP/polyunsaturated FA monoglyceride (polyunsaturated MG)/polyunsaturated FA, SLP/medium chain FA, and SLP/polyunsaturated FA, SLP/medium chain MG, and SLP/polyunsaturated MG also showed a high degree of activity. Wettability decreased rapidly when the amount of saturated, long chain FA moieties increased. It is recognized that the degrees of unsaturation and the chain length of FAs in the lipid mixtures have a decisive influence on surface activities. Higher ratios of MG and FA to SLP gave higher activity; and solubilizers such as bile salts were necessary to dissolve them in water.     

酶解时间对大豆蛋白-溶血磷脂相互作用及乳化特性的影响

通过不同酶解时间得到大豆溶血磷脂,对大豆分离蛋白-溶血磷脂相互作用及其对复合乳化体系乳化特性的影响进行探究,采用荧光光谱法在Stern-Volmer和Van’t Hoff方程基础上对大豆分离蛋白-溶血磷脂荧光猝灭作用、相互结合常数、结合位点及相互作用力类型进行判断,并对复合乳化体系分别进行乳化活性、乳化稳定性的测定及微观结构变化的观察。结果表明:随着磷脂酶A1酶解时间的延长,大豆分离蛋白-溶血磷脂相互作用先增强后下降,乳化特性指标同样基本呈现先升高后降低的趋势,这表明二者的相互作用对乳化特性具有一定影响。其中,当酶解时间为4 h时,二者相互作用最强,乳液的乳化特性表现最佳,这表明适度酶解产生的溶血磷脂会促进其与大豆分离蛋白的相互作用,在水油界面上形成较稳定的界面膜,形成稳定的复合乳状液。     

Phospholipases a(1)

Phospholipase A(1) (PLA(1)) is an enzyme that hydrolyzes phospholipids and produces 2-acyl-lysophospholipids and fatty acids. This lipolytic activity is conserved in a wide range of organisms but is carried out by a diverse set of PLA(1) enzymes. Where their function is known, PLA(1)s have been shown to act as digestive enzymes, possess central roles in membrane maintenance and remodeling, or regulate important cellular mechanisms by the production of various lysophospholipid mediators, such as lysophosphatidylserine and lysophosphatidic acid, which in turn have multiple biological functions.     

Lysophosphatidylinositol Induced Morphological Changes and Stress Fiber Formation through the GPR55-RhoA-ROCK Pathway

We previously reported that lysophosphatidylinositol (LPI) functions as an endogenous agonist of GPR55, a novel cannabinoid receptor. However, the physiological roles of LPI-GPR55 have not yet been elucidated in detail. In the present study, we found that LPI induced morphological changes in GPR55-expressing HEK293 cells. LPI induced the cell rounding of GPR55-expressing HEK293 cells but not of empty-vector-transfected cells. LPI also induced the activation of small GTP-binding protein RhoA and increased stress fiber formation in GPR55-expressing HEK293 cells. The inhibition of RhoA and Rho kinase ROCK by the C3 exoenzyme and the ROCK inhibitor reduced LPI-induced cell rounding and stress fiber formation. These results clearly indicated that the LPI-induced morphological changes and the assembly of the cytoskeletons were mediated through the GPR55-RhoA-ROCK pathway.     

Effects of lysophospholipids on short-term production,nitrogen utilization,and rumen fermentation and bacterial population in lactating dairy cows

An experiment was conducted to examine effects of supplemental lysophospholipids (LPL) in dairy cows. Eight ruminally cannulated lactating Holstein cows were used in a replicated 4 × 4 Latin square design. Dietary treatments were (1) a dairy ration [CON; 55% forage and 45% concentrate on a dry matter (DM) basis], (2) a positive control diet supplemented with monensin (MON; 16 mg/kg in dietary DM; Elanco Animal Health, Greenfield, IN], (3) a control diet supplemented with low LPL (0.05% of dietary DM; Lipidol Ultra, Easy Bio Inc., Seoul, South Korea), and (4) a control diet supplemented with high LPL (0.075% of dietary DM). Experimental periods were 21 d with 14-d diet adaptation and 7-d sample collection. Daily intake and milk yield were measured and rumen contents were collected for fermentation characteristics and bacterial population. Spot urine and fecal samples (8 samples/cow per period) were collected to determine nutrient digestibility and dietary N utilization. All data were analyzed using the MIXED procedure of SAS (SAS Institute Inc., Cary, NC; group and cow within group were random effects and treatments, time, and their interaction were fixed effects). Preplanned contrasts were made to determine effect of MON versus CON, effect of LPL versus MON, and linear effect of increasing LPL. In the current study, responses to MON generally agreed with effects of monensin observed in the literature (increased milk yield and feed efficiency but decreased milk fat content). Supplementation of LPL to the diet did not alter DM intake but linearly increased milk yield, resulting in increases in feed efficiency (milk yield/DM intake) and milk protein and fat yields. However, total-tract digestibility of DM and organic matter tended to be lower (60.9 vs. 62.2% and 61.8 vs. 63.1%, respectively) for LPL compared with CON. Linear increases in milk N secretion and decreases in urinary N excretion were observed with increasing LPL in the diet. A slight decrease in acetate proportion in the rumen for LPL was found. Relative to MON, very few bacteria in the rumen were affected with increasing LPL. In conclusion, LPL is a potential feed additive that can increase milk yield and components and dietary N utilization. However, more studies with large numbers of animals are needed to confirm the effect of LPL on production. Similar positive effects on production were observed between LPL and MON, but individual mechanisms were likely different according to ruminal fermentation characteristics. Further studies are needed to explore the mode of action of LPL in dairy cows.     

磷脂酶A2水解菜籽油脚的研究

油脚中含有丰富的磷脂,磷脂的酶改性产物溶血磷脂有极广的应用。以菜籽油脚为底物,以实验室自制的磷脂酶A2为水解酶,采用水相反应方式,通过单因素实验和多因素正交实验,分别对底物用量、Ca2 添加量、反应温度、起始pH等因素进行了系统的研究。结果表明,菜籽油脚酶解的最佳条件为:底物浓度20%,钙离子浓度0.3%,温度50℃,起始pH8.0。在最佳条件下,FFA生成量可达到172.3μmol/g。     

脂肪酶Bakezyme LFP水解大豆粉末磷脂研究

以酸值为指标,通过单因素实验和正交实验确定了脂肪酶Bakezyme LFP在水相中水解大豆粉末磷脂的最佳工艺条件为:反应温度为50℃,反应时间为8h,p H为5.0,加酶量为8%,底物质量浓度为25mg/m L。在该工艺条件下,大豆磷脂水解产物酸值为65.2mg KOH/g。利用气相色谱-质谱联用仪分析了水解前后大豆磷脂的脂肪酸组成,结果表明水解后饱和脂肪酸含量明显下降,不饱和脂肪酸含量明显增加,说明脂肪酶Bakezyme LFP主要水解大豆磷脂1位上脂肪酸,具有磷脂酶A1的特性。利用高效液相色谱分析水解前后大豆磷脂的磷脂酰乙醇胺（PE）和磷脂酰胆碱（PC）含量,结果表明大豆磷脂中PE、PC发生水解,生成溶血磷脂。