Effects of branched-chain volatile fatty acids on lactation performance and mRNA expression of genes related to fatty acid synthesis in mammary gland of dairy cows

Branched-chain volatile fatty acids (BCVFA) supplements could promote lactation performance and milk quality by improving ruminal fermentation and milk fatty acid synthesis. This study was conducted to evaluate the effects of BCVFA supplementation on milk performance, ruminal fermentation, nutrient digestibility and mRNA expression of genes related to fatty acid synthesis in mammary gland of dairy cows. A total of 36 multiparous Chinese Holstein cows averaging 606±4.7 kg of BW, 65±5.2 day in milk (DIM) with daily milk production of 30.6±0.72 kg were assigned to one of four groups blocked by lactation number, milk yield and DIM. The treatments were control, low-BCVFA (LBCVFA), medium-BCVFA (MBCVFA) and high-BCVFA (HBCVFA) with 0, 30, 60 and 90 g BCVFA per cow per day, respectively. Experimental periods were 105 days with 15 days of adaptation and 90 days of data collection. Dry matter (DM) intake tended to increase, but BW changes were similar among treatments. Yields of actual milk, 4% fat corrected milk, milk fat and true protein linearly increased, but feed conversion ratio (FCR) linearly decreased with increasing BCVFA supplementation. Milk fat content linearly increased, but true protein content tended to increase. Contents of C4:0, C6:0, C8:0, C10:0, C12:0, C14:0 and C15:0 fatty acids in milk fat linearly increased, whereas other fatty acids were not affected with increasing BCVFA supplementation. Ruminal pH, ammonia N concentration and propionate molar proportion linearly decreased, but total VFA production and molar proportions of acetate and butyrate linearly increased with increasing BCVFA supplementation. Consequently, acetate to propionate ratios linearly increased. Digestibilities of DM, organic matter, CP, NDF and ADF also linearly increased. In addition, mRNA expressions of peroxisome proliferator-activated receptor γ, sterol regulatory element-binding factor 1 and fatty acid-binding protein 3 linearly increased, mRNA expressions of acetyl-coenzyme A carboxylase-α, fatty acid synthase and stearoyl-CoA desaturase quadratically increased. However, lipoprotein lipase mRNA expression was not affected by treatments. The results indicated that lactation performance and milk fat synthesis increased with BCVFA supplementation by improving ruminal fermentation, nutrient digestibility and mRNA expressions of genes related to milk fat synthesis.     

Modelling on isoamyl isovalerate synthesis from Rhizomucor miehei lipasein organic media: optimization studies

Immobilized lipase from Rhizomucor miehei (Lipozyme IM-20) was employed in the esterification of isovaleric acid and isoamyl alcohol to synthesize isoamyl isovalerate in n-heptane. Response surface methodology (RSM) based on a five-level, five-variable central composite rotatable design (CCRD) was used to evaluate the effects of important variables: enzyme concentration (20–40% w/w of acid), acid concentration (0.2–1.0 M), incubation period (24–120 h), alcohol concentration (0.25–1.25 M) and temperature (30–70 °C) on the esterification yield of isoamyl isovalerate. Extent of conversion was found to be excellent at all acid and alcohol concentrations employed in the range of 0.2–1.25 M, even at low enzyme concentration (20% w/w). The optimum conditions arrived at are as follows: 35% (w/w) enzyme concentration, 1.0 M acid concentration, 1.25 M alcohol concentration and 120 h incubation period, at 35 °C. Under these conditions, the predicted value was 680 mM ester matched very well with an experimental value of 678 mM.     

Peculiarities of Ethanol Metabolism in an Acinetobacter sp. Mutant Strain Defective in Exopolysaccharide Synthesis

Activities of the key enzymes of ethanol metabolism were assayed in ethanol-grown cells of an Acinetobacter sp. mutant strain unable to synthesize exopolysaccharides (EPS). The original EPS-producing strain could not be used for enzyme analysis because its cells could not to be separated from the extremely viscous EPS with a high molecular weight. In Acinetobacter sp., ethanol oxidation to acetaldehyde proved to be catalyzed by the NAD⁺-dependent alcohol dehydrogenase (EC 1.1.1.1.). Both NAD⁺ and NADP⁺ could be electron accepters in the acetaldehyde dehydrogenase reaction. Acetate is implicated in the Acinetobacter sp. metabolism via the reaction catalyzed by acetyl-CoA-synthetase (EC 6.2.1.1.). Isocitrate lyase (EC 4.1.3.1.) activity was also detected, indicating that the glyoxylate cycle is the anaplerotic mechanism that replenishes the pool of C₄-dicarboxylic acids in Acinetobacter sp. cells. In ethanol metabolism by Acinetobacter sp., the reactions involving acetate are the bottleneck, as evidenced by the inhibitory effect of sodium ions on both acetate oxidation in the intact cells and on acetyl-CoA-synthetase activity in the cell-free extracts, as well as by the limitation of the C₂-metabolism by coenzyme A. The results obtained may be helpful in developing a new biotechnological procedure for obtaining ethanol-derived exopolysaccharide ethapolan.     

Effects of particle size and drying methods of corn on growth performance,digestibility and haematological and immunological characteristics of weaned piglets

The objective of this study was to evaluate the effects of particle size and drying methods of corn on growth performance of weaned piglets. Crossbreed weaned piglets (n = 192; Duroc × Landrace × Large White) were assigned to one of four treatments (2 × 2 factorial arrangement). All piglets were fed corn–soybean meal diets and treatments were (1) hot air-dried and coarsely ground corn, (2) hot air-dried and finely ground corn, (3) sun-dried and coarsely ground corn and (4) sun-dried and finely ground corn. The results showed that finely ground corn (FGC) improved the performance of piglets. Additionally, the apparent total tract digestibility (ATTD) of gross energy (GE) and ether extract (EE) were increased by FGC, but the drying methods did not affect the performance of piglets or ATTD. Furthermore, smaller particle size significantly decreased the intestinal permeability, which was also not influenced by drying methods. FGC increased the total number of white blood cells, but not other blood parameters. Finally, the level of serum interleukin-1 was decreased by fine grinding and that of serum tumour necrosis factor α was decreased by sun drying. Conversely, these characteristics of weaned piglets can hardly have been affected either by the corn drying method or its interaction with grinding methods.     

腾冲嗜热厌氧菌丙氨酸消旋酶底物通道氨基酸位点的功能

【目的】通过定点突变探究腾冲嗜热厌氧菌MB4中生物合成型丙氨酸消旋酶Tt Alr底物通道内氨基酸位点A172和S173的功能。【方法】利用定点突变PCR技术构建突变体,通过亲和层析法纯化酶蛋白,采用D-氨基酸氧化酶偶联法检测各突变蛋白的活性及其稳定性。【结果】通过定点突变PCR成功得到8个突变体,酶学特性分析发现,A172位点突变为丝氨酸(S)后酶蛋白的相对活性有所提升,但含有该位点突变的酶蛋白稳定性均大幅下降;S173位点突变为天门冬氨酸(D)后导致突变体蛋白的最适反应温度提升了15°C,半衰期大幅延长,但相对活性明显下降。【结论】丙氨酸消旋酶Tt Alr底物通道内A172和S173位点均是影响酶蛋白催化活性和稳定性的关键位点。     

不同酶对草鱼蛋白功能特性及风味成分的影响

通过蛋白酶对草鱼进行酶解,对其酶解产物的功能特性和酶解液的风味成分进行研究。结果显示,风味蛋白酶、胰蛋白酶酶解产物的溶解性、热稳定性均优于胃蛋白酶酶解产物(P0.05)。经酶处理后鱼肉酶解液鲜甜味氨基酸占比增加,苦味氨基酸占比减少;利用SPME-GC-MS共检测出83种挥发性成分,以醛酮类和醇类为主,酶解处理后醇类的相对含量显著减少,醛酮类的相对含量则显著增加,酶解液具有独特风味。经胰蛋白酶、风味蛋白酶酶解后的草鱼肉酶解产物及酶解液均可作为食品基料应用于加工中。     

Effects of isobutyrate on rumen fermentation,urinary excretion of purine derivatives and digestibility in steers

The objective of this study was to evaluate the effects of isobutyrate supplementations on rumen fermentation, urinary excretion of purine derivatives and feed digestibility in steers. Eight ruminally cannulated Simmental steers were used in a replicated 4 × 4 Latin square experiment. On DM basis, diet consisted of 60% corn stover and 40% concentrate. Dry matter intake (averaged 9 kg/d) was restricted to 90% of ad libitum intake. The four treatment groups received a daily dose of 0 (control), 8.4, 16.8 or 25.2 g isobutyrate per steer. With increasing isobutyrate supplementation total VFA concentration (range 64.2–74.0 mM) was significantly enhanced. The ratio of acetate to propionate (range 2.72–4.25) was also significantly increased due to the increase in actate production and decrease in propionate production. With increasing isobutyrate supplementation the ruminal degradation of NDF from corn stover was improved but the CP degradability of soybean meal was decreased. Furthermore, the isobutyrate supplementation caused a significantly increased urinary excretion of purine derivatives. Similarly, digestibilities of OM, NDF and CP in the total tract were significantly increased. The present results indicate that dietary supplementation with isobutyrate improved rumen fermentation and feed digestion in beef cattle in a dose-dependent manner. According to the conditions of this experiment, the optimum daily dose of isobutyrate was about 16.8 g/animal.     

酶法提取银杏黄酮类化合物研究

本文研究了纤维素酶酶解法提取银杏总黄酮工艺。与传统的乙醇提取工艺相比,银杏总黄酮得率提高了18．92％。实验确定了最佳提取条件：酶浓度0．40mg／mL,酶作用时间120min,酶解温度50℃,酶解介质pH值为4．5,乙醇浓度70％,提取温度70℃。     

Thermal stability and enzymatic activity of a smaller lysozyme from silk moth (Bombyx mori)

Bombyx mori lysozyme is 10 amino acids shorter than hen egg-white lysozyme, which is a typical c-type lysozyme. It was expressed by using the methylotrophic yeast Pichia pastoris. The thermal stability and the enzymatic activity of the Bombyx mori lysozyme were estimated and compared with those of human and hen egg-white lysozymes. The denaturation temperature was 17-26°C lower than those of human and hen egg-white lysozymes. Further, the enthalpy change and the heat capacity change for unfolding were smaller than those of human lysozyme. It was also confirmed that the stability against guanidine hydrochloride was lower than those of the other two lysozymes. The enzymatic activity toward a simple synthetic substrate was measured and compared with those of human and hen egg-white lysozymes. The B-F binding mode was obviously dominant, although the A-E binding mode was preferred in human and hen egg-white lysozymes.