Physicochemical properties of the thermal gel of water-washed meat in the presence of the more soluble fraction of porcine sarcoplasmic protein

We investigated the physicochemical properties of the thermal gel of water‐washed pork meat (WWM) in the presence of the soluble fraction of porcine sarcoplasmic protein (SP) obtained with ammonium sulfate at 75 percent saturation. Two precipitated fractions of SP were obtained at 0–50 percent and 50–75 percent saturation, named SP‐f1 and SP‐f2, respectively, and the soluble fraction obtained at 75 percent saturation, SP‐f3, was used. Sodium dodecyl sulfate‐polyacrylamide gel electrophoresis showed that SP‐f3 contained mainly glyceraldehyde‐3‐phosphate dehydrogenase (GAPDH), while SP‐f1 and SP‐f2 had other SPs such as phosphorylase b, enolase, actin and phosphoglycerate mutase. The gel strength of WWM was greater when SP‐f3 rather than one of various animal proteins such as bovine plasma (BP), egg white, or whey protein isolates (WPI), was added and SP‐f3 had a gel‐enhancing effect as good as that of polyphosphate (PP). The gel strength of WWM with added SP‐f3 increased significantly with NaCl at 0.15 mol/L or more, but not in the absence of NaCl (0 mol/L). The effect of SP‐f3 was evident at neutral pH and maximum gel strength was obtained at a pH above 6.0. Differential scanning calorimetric (DSC) analysis showed that an endothermic peak corresponding to myosin heads in WWM shifted to a lower temperature with the addition of SP‐f3, as in the case of PP, though there was no such shift in the presence of other animal proteins (BP, egg white and WPI), suggesting that SP‐f3 increases the gel strength of WWM through the dissociation of actomyosin similar to PP. Scanning electron microscopy (SEM) revealed wall‐like structures among the protein strands in the WWM gel matrix in the presence of SP‐f3. The results of DSC and SEM indicated that the formation of a gel network in meat products is reinforced with GAPDH in SP after the interaction between GAPDH and myofibrillar protein.     

三种鹿血清乳酸脱氢酶同功酶的电泳研究

采用聚丙烯酰胺凝胶电泳法对26头白唇鹿、41头马鹿和11头梅花鹿的血清乳酸脱氢酶（LDH）同功酶谱及其多态性进行了研究。结果发现：三种鹿的LDH都有LDH1、LDH2、LDH3和LDH5四种同功酶;LDH3同功酶具有1～3条亚带,LDH5具有显现酶活性和不显现酶活性两种表型而呈现多态性;白唇鹿的LDH同功酶酶谱及其多态性有较明显的种的特征。     

双向电泳结合蛋白质印迹技术分析猪囊尾蚴乳酸脱氢酶

为了研究猪囊尾蚴乳酸脱氢酶(LDH)的表达,试验选择四川省雅江县呷拉乡猪带绦虫病患者,给其口服槟榔-南瓜子驱虫,收集、制备虫卵悬液(8万个/mL),再给3头20日龄三元杂交乳猪猪灌胃虫卵悬液,每头1 mL,40 d后收集、制备猪囊尾蚴蛋白,进行双向电泳(2-DE)分析,将凝胶蛋白斑点转移至聚偏氟乙烯膜(PVDF膜),用自制的大鼠抗猪带绦虫LDH血清作为一抗、健康大鼠血清作为阴性对照进行蛋白质印迹(Western-blotting)分析。结果表明:双向电泳凝胶共检测到(207±9)个蛋白质斑点,相对分子质量(Mr)为14 400～94 000,等电点(pI)为3.0～10.0;试验组特异性抗原抗体阳性杂交斑点为1个,阴性对照未见阳性杂交斑点;将Western-blotting检测的抗原抗体阳性杂交斑点与原双向电泳凝胶斑点进行比对,找到对应蛋白斑点,经ImageMaster 2D Platinum 5.0软件分析后初步确定该蛋白斑点的pI/Mr为7.03/35 368,与猪带绦虫LDH的pI/Mr理论推导值接近,说明猪囊尾蚴表达LDH。     

Gene expression of fatty acid-binding proteins, fatty acid transport proteins (cd36 and FATP) and β-oxidation-related genes in Atlantic salmon (Salmo salar L.) fed fish oil or vegetable oil

Relative gene expression pattern of fatty acid transport proteins (FATP and cd36), intracellular fatty acid-binding proteins (FABP3, FABP10 and FABP11), β-oxidation-related genes [carnitine palmitoyl transferase II (CPTII), peroxisome proliferator-activated receptor β (PPARβ), acyl-CoA oxidase (AOX), long-chain fatty acyl-CoA synthetase (FACS), acyl-CoA dehydrogenase (dehydrogenase)] and uncoupling protein 2 (UCP2) was assessed by RT-qPCR in Atlantic salmon muscle (red and white), liver, heart, myosepta and visceral fat. FABP11, a FABP isoform not previously described in Atlantic salmon, was highly expressed in visceral fat and myosepta and at the lower level in red muscle, white muscle, myosepta and heart. Furthermore, Atlantic salmon were fed either a diet containing fish oil (FO) or a complete replacement of FO with a vegetable oil blend (55% rapeseed oil, 30% palm oil and 15% linseed oil; VO) for the production cycle (27 months from start of feeding and until ∼4.5 kg mean weight). The expression of genes related to β-oxidation, fatty acid uptake and transport in the white muscle indicate ( n  = 3) significant down-regulation in VO fed Atlantic salmon and correlated with previously reported white muscle triacylglycerol stores and β-oxidation. FABP11 in visceral fat and myosepta was also down-regulated in VO fed fish.     

羊草乙醛脱氢酶（ALDH）基因片段的克隆及表达分析

[研究目的]克隆羊草的乙醛脱氢酶ALDH基因片段,研究该基因在不同条件下的表达情况。[方法]采用RT-PCR技术克隆羊草的ALDH基因片段,并对其核苷酸和氨基酸序列进行分析,采用RealTimeRT-PCR方法研究该基因的表达。[结果]获得了羊草的ALDH基因片段,长度为675bp,编码225aa。核苷酸序列比较表明,与水稻ALDH1a序列(AB037421)同源性为86%,与玉米RF2C(AF348413)同源性为85%。BLASTp分析,该序列与水稻、玉米、拟南芥的乙醛脱氢酶一致性分别高达87%、86%、60%,含有醛脱氢酶基因家族的保守结构域。RealTimeRT-PCR数据表明,在诱导条件下该基因的表达量呈先升高后降低的趋势。总体上来说,该基因对盐的响应要高于干旱和冷冻。[结论]成功获得了羊草ALDH基因片段,并研究了该基因的表达情况,为进一步克隆羊草ALDH全长基因奠定了基础。     

Stereochemistry of matairesinol formation by <Emphasis Type="Italic">Daphne</Emphasis> secoisolariciresinol dehydrogenase

Secoisolariciresinol dehydrogenase activity was detected for the first time from Daphne odora and Daphne genkwa (Thymelaeaceae), which are known to produce optically pure (+)-matairesinol. In sharp contrast, (–)-matairesinol was formed selectively over the (+)-antipode by the secoisolariciresinol dehydrogenase preparation from both D. odora callus and D. genkwa shoots.     

逆境胁迫下苜蓿乙醛脱氢酶基因的表达

为研究苜蓿植株在盐、低温、干旱胁迫下,植株体内的乙醛脱氢酶（ALDH）基因表达量的变化,在3种逆境条件下分别取处理0、4、8、12、24h叶片,用荧光定量RT—PCR法测定了ALDH基N的表达量。结果表明,与对照相比,盐胁迫的苜蓿植株叶片ALDH基因表达量随处理时间的不同,表达量有所上升也有所下降;低温胁迫处理的植株叶片ALDH基因随处理时间的不同表达量有升有降;干旱胁迫处理下,对干旱的胁迫处理敏感性很差,表达量很低。     

半胱胺对藏绵羊断奶羔羊瘤胃pH值、总脱氢酶及TVFA的影响

试验研究了半胱胺(Cs)对藏绵羊断奶羔羊瘤胃pH值、总脱氢酶及TVFA的影响。将10只(公母各半)80日龄左右、体重平均为(16.00±0.31)kg、装有永久性瘤胃瘘管的藏绵羊断奶羔羊随机分为2组(试验组和对照组),在日粮精料中添加300 mg/(kg·BW)的Cs,隔日添加1次,试验期42 d。正式试验当日及试验第7、14、21、28、35天的7:00、9:00、11:00、13:00、15:00、17:00、19:00、21:00、23:00、1:00、3:00、5:00于颈静脉采血样10 mL,同时采集瘤胃液10 mL,测定瘤胃pH值、总脱氢酶的活性和TVFA。检测结果表明,试验组精料中添加Cs使藏绵羊断奶羔羊瘤胃液pH值升高;血清尿素氮浓度降低,与对照组相比差异极显著(P0.01);瘤胃液总脱氢酶活力提高极其明显(P0.01);整个试验期内,半胱胺对瘤胃TVFA的影响极显著(P0.01)。     

Water quality affects SDH activity, protein content and RNA:DNA ratios in fish (Catla catla, Labeo rohita and Oreochromis mossambicus) raised in ponds of a sewage-fed fish farm

The spatial and seasonal variability of the respiratory enzyme succinate dehydrogenase and the protein content were examined in different tissues of fish cultured in three ponds along the effluent gradient of a sewage-fed fish farm. Indian major carp Catla catla (150-230 g) and Labeo rohita (60-190 g) cultured in both the middle and last points of the sewage effluent (stocking pond 1) and (stocking pond 4) and Oreochromis mossambicus (50-160 g), a naturally growing fish of the inlet (facultative pond) and the out let of the sewage effluent (stocking pond 4) were procured every month during the period of January-December, 2005 and were subjected to determination of succinate dehydrogenase activity, total protein, DNA and RNA contents from gill, liver and muscle tissue respectively. The SDH activity of all three test fishes (Catla catla, Labeo rohita and Oreochromis mossambicus) was reduced significantly (ANOVA; P < 0.05) when cultured in SP-4 compared to SP-1 in the case of Catla catla and Labeo rohita and in facultative pond in the case of Oreochromis mossambicus.Conspicuous differences in the SDH activity of fish between the last and first stocking pond or the facultative pond were clearly due to the result of the differences in water quality. There was a direct relationship between SDH activity in gill tissue of any of the fish investigated and ammonia-N concentration of water or water pH. This shows that the respiratory activity of these fishes was strongly affected by the ammonia and pH of water. In other words, this suggests that as the distance from the point source increases, there was a substantial improvement of water quality in the ponds located along the sewage effluent gradient. Evidently, there is a progressive pattern of growth, survival and physiological health of fish and abundance of favorable diversity of food organisms with rich biodiversity.     

Long-term nutritional effects on the primary liver and kidney metabolism in rainbow trout (Oncorhynchus mykiss). II. Adaptive response of glucose 6-phosphate dehydrogenase activity to high-carbohydrate/low-protein and high-fat/non-carbohydrate diets

The influence of long-term administration of high-carbohydrate/low-protein and high-fat/non-carbohydrate diets were studied in relation to kinetic behaviour of glucose 6-phosphate dehydrogenase in liver and kidney of rainbow trout. In all cases studied, the saturation curves of these enzyme showed typical hyperbolic kinetics without evidence of sigmoidicity. After 30 days of feeding with a high-fat diet (170 g kg^?1), there was a significant decrease in V_max and specific activity (45%) as well as catalytic efficiency (39%) without changes in K_m or activity ratio of hepatic glucose 6-phosphate dehydrogenase. These changes agree more with a clearly decreased cell concentration than with an inhibition of the pre-existing enzyme. The administration of a high-carbohydrate diet (60 g kg^?1), contrary to what was previously thought, decreased V_max by 21% and specific activity and catalytic efficiency by 30%, without significant changes in the other kinetic parameters of the hepatic enzyme. The kinetic behaviour under these nutritional conditons was due to the rejection of this diet by the fish and thus could be considered a low-feeding situation. On the other hand, no variations in the kinetics of renal glucose 6-phosphate dehydrogenase were found, clearly demonstrating that in this organ, the pentosephosphate pathway showed no adaptive response related to fattyacid and other lipid synthesis. The activity of the renal enzyme was consistently half that of the hepatic enzyme.