冰淇淋混合料液的老化

阐述了冰淇淋混合料液老化过程的条件控制及混合料在老化过程中发生的主要变化。经过老化过程,混合料液的黏度显著增加,有助于凝冻过程中的搅打起泡,使冰淇淋产品的质构更加细腻,抗融性和贮藏稳定性更好。     

抗凋亡融合蛋白PTD-Bcl-xL原核表达载体的构建及表达纯化

人工抗凋亡蛋白PTD-Bcl-x L能保护多种因素引起的细胞异常凋亡,为了获得高纯度Bcl-x L与PTD(Protein transduction domains)的融合蛋白,首先采用TRIzol法提取SD大鼠肝脏总RNA,将RNA反转录为c DNA,设计引物以c DNA为模板,PCR扩增Bcl-x L基因,构建p UM19-T-Bcl-x L质粒,并对质粒双酶切鉴定和测序鉴定;其次设计包含PTD序列的Bcl-x L引物,以测序正确的p UM19-T-Bcl-x L质粒为模板,PCR扩增PTD-Bcl-x L序列,将扩增序列克隆入p ET28a载体,构建PTD-Bcl-x L蛋白的原核表达质粒p ET28a-PTD-Bcl-x L,并对p ET28a-PTD-Bcl-x L载体双酶切鉴定和测序鉴定;将p ET28a-PTD-Bcl-x L重组质粒转化大肠杆菌BL21(DE3),用IPTG诱导表达,并对IPTG诱导融合蛋白表达的浓度和诱导时间进行了优化;SDS-PAGE分析表达蛋白的可溶性情况,在变性条件下用Ni-NTA琼脂纯化融合蛋白;最后用SDS-PAGE、Western Blot及质谱对融合蛋白进行鉴定。结果表明:双酶切p UM19-T-Bcl-x L质粒出现约774 bp大小条带,p UM19-T-Bcl-x L质粒测序结果与NCBI数据库比对序列一致,表明成功构建p UM19-T-Bcl-x L质粒;双酶切p ET28a-PTD-Bcl-x L质粒出现约744 bp大小条带,p ET28a-PTD-Bcl-x L质粒测序结果与预期序列一致,表明成功构建了p ET28a-PTD-Bcl-x L原核表达载体;在IPTG诱导下p ET28a-Bcl-x L重组质粒在大肠杆菌BL21(DE3)中表达出36 k Da大小蛋白,最优IPTG诱导浓度为0.1 mmol/L,最佳IPTG诱导时间为6 h;SDS-PAGE电泳显示融合蛋白主要出现在菌液超声后的沉淀里,以包涵体形式表达,经Ni-NTA琼脂纯化获得了高纯度的融合蛋白;Western Blot和质谱鉴定证明IPTG诱导表达蛋白和纯化的融合蛋白为PTD-Bcl-x L蛋白。纯化得到了PTD-Bc L-x L融合蛋白,推进了PTD-Bcl-x L蛋白在猪、牛等家畜精液冷冻保存的应用进程。     

陆地棉ZF-HD蛋白的全基因组分析

ZF-HD(Zinc finger-homeodomain)蛋白属于同源异型盒蛋白家族,在动植物发育过程中起重要的作用。利用生物信息学的手段,研究陆地棉标准系TM-1基因组中ZF-HD蛋白的数目、亚细胞定位、染色体分布、基序、进化关系和组织表达情况。TM-1中共有35个Gh ZHD蛋白成员,且大部分成员定位到细胞核内;分布在20条染色体和2条Scaffold上,且具有11对直系同源基因;进化树分析表明,TM-1基因组中的ZF-HD蛋白大致分为6个小组,每个小组成员间具有类似的基序类型和排列顺序;大部分Gh ZHD基因在胚珠和纤维组织中表达,还有部分基因在根、茎、花、蕾和分生区中表达,在叶和愈伤组织中表达的基因最少。     

红素氧还蛋白研究进展

自然界中大多数的红素氧还蛋白(rubredoxin)集中来自严格厌氧菌,包括细菌、古菌和少数微需氧细菌,而植物中该类蛋白相对较少,它的功能主要表现在作为辅因子或电子供体参与多个生化途径。也有研究显示,无论是植物中还是微生物中的红素氧还蛋白基因在协助生物抵御氧化胁迫方面均发挥着重要的作用。本文全面介绍自然界中的红素氧还蛋白基因的分布、结构以及在微生物、植物中所发挥的作用等,希望能为今后的进一步研究提供参考。     

马乳脂肪球膜蛋白组鉴定及潜在功能分析

试验旨在研究马乳脂肪球膜（milk fat globule membrane，MFGM）蛋白组成及潜在的生物学功能。选取12匹5岁左右、平均体重为450~500 kg的健康纯血母马作为试验动物，主要饲喂干牧草，所有马匹饲养和管理条件均相同。将12份样品以每组4份进行混合，分离提取产后第60天马乳中MFGM蛋白，进行高分辨质谱仪鉴定和生物信息学分析。结果显示，马MFGM组分中共鉴定出310种表达蛋白，这些蛋白主要参与的生物过程为生物调节、刺激应答、定位、多细胞生物过程、运输、信号、细胞通讯、发育过程、细胞分化和免疫系统过程等；细胞成分主要为胞外区域、膜、囊泡、核仁和线粒体等；分子功能主要为催化活性、蛋白质结合、碳水化合物衍生物结合和小分子结合等。KEGG通路分析表明，MFGM蛋白主要参与血小板活化通路、内吞、脂肪酸生物合成和Ras信号通路等。马乳MFGM蛋白的蛋白质互作网络分析图中共包含215种蛋白质。本研究结果揭示了马乳MFGM蛋白质组的复杂性，为解析其营养和生物学功能提供了科学数据。     

Effect of wilting,silage additive,PEG treatment and tannin content on the distribution of N between different fractions after ensiling of three different sainfoin (Onobrychis viciifolia) varieties

Sainfoin (Onobrychis viciifolia) is a tanniniferous, leguminous plant that has potentially beneficial effects on protein utilization in ruminants. As ensiling causes protein breakdown and elevated levels of buffer soluble N (BSN), we studied the distribution of N before and after ensiling sainfoin. Three varieties of sainfoin were either direct‐cut and frozen directly or wilted and frozen before later ensiling in mini‐silos with and without acidification with Promyr (PM; an acidifying commercial mixture of propionic and formic acid) and with or without polyethylene glycol (PEG). Extractable tannins (ET) and protein‐bound tannins (PBT) were measured with an HCl/butanol method in an attempt to correlate tannin levels to N fractions. The sainfoin silages showed good ensiling characteristics and had relatively high concentrations of undegraded protein. The effect of wilting on BSN levels (g/kg N) was dependent on sainfoin variety (P < 0·001). PEG increased and PM decreased the level of BSN in the silages (P < 0·001). PM treatment also produced less non‐protein N and ammonia‐N (P < 0·05) as compared with no additive. Addition of PEG to the silage increased the BSN‐proportion 1·8‐ and 2·6‐fold for both DM stages. A strong tannin‐protein binding effect is, therefore, confirmed in sainfoin. However, correlations between tannin levels (ET and PBT) and BSN were poor in the (non‐PEG) silages, indicating either that the HCl/butanol method is unsuitable for measuring tannin in silages or that qualitative attributes of tannins are more relevant than quantitative. The HCl/butanol method seems therefore not to be useful to predict degradation of protein in sainfoin silages.     

利用甜高粱秸秆汁产单细胞蛋白的分批补料发酵研究

本文以甜高粱秸秆汁为主要的发酵原料,对热带假丝酵母SG-1在15-L罐上产单细胞蛋白(SCP)的分批补料发酵过程进行研究。首先对分批发酵模式下的菌体代谢过程变化进行研究。研究发现,接种后菌体快速进入繁殖阶段,但是总糖在12 h处于耗竭状态,使得菌体量和SCP产量过早的在18 h就达到峰值。基于此,在15-L罐上建立了热带假丝酵母SG-1产SCP的分批补料发酵模式。结果表明:菌体代谢因基质的补加而得到进一步的延续,放罐时(42 h)其菌体干重和SCP产量分别达19.86 g/L和9.93 g/L,显著高于分批发酵模式的10.45 g/L和4.70 g/L。     

Dietary fiber in a low-protein diet during gestation affects nitrogen excretion in primiparous gilts,with possible influences from the gut microbiota

We investigated the effects of dietary fiber (DF) supplementation in normal or low crude protein (CP) diets on reproductive performance and nitrogen (N) utilization in primiparous gilts. In total, 77 Landrace × Yorkshire pregnant gilts were randomly allocated to four dietary treatments in a 2 × 2 factorial design. The groups comprised 1) equal intake of normal CP (12.82% and 0.61% total lysine), 2) low CP (LP) (10.53% and 0.61% total lysine), and 3) with or 4) without DF supplementation (cellulose, inulin, and pectin in a 34:10:1 ratio). A low-protein diet during gestation significantly reduced daily weight gain from days 91 to 110 of pregnancy (−162.5 g/d, P = 0.004). From N balance trials conducted at days 35 to 38, 65 to 68, and 95 to 98 of pregnancy, DF addition increased fecal N excretion at days 65 to 68 (+24.1%) and 95 to 98 (+13.8%) of pregnancy (P < 0.05) but reduced urinary N excretion (P < 0.05), resulting in greater N retention at each gestational stage. DF increased fecal microbial protein levels and excretion during gestation. An LP diet also reduced urinary N excretion at different gestational stages. An in vitro fermentation trial on culture media with nonprotein N urea and ammonium bicarbonate (NH₄HCO₃) as the only N sources revealed that microbiota derived from feces of gestating gilts fed the high DF diet exhibited a greater capacity to convert nonprotein N to microbial protein. Microbial fecal diversity, as measured by 16S rRNA sequencing, revealed significant changes from DF but not CP diets. Gilts fed an LP diet had a higher number of stillbirths (+0.83 per litter, P = 0.046) and a lower piglet birth weight (1.52 vs. 1.37 kg, P = 0.006), regardless of DF levels. Collectively, DF supplementation to gestation diets shifted N excretion from urine to feces in the form of microbial protein, suggesting that the microbiota had a putative role in controlling N utilization from DF. Additionally, a low-protein diet during gestation negatively affected the litter performance of gilts.     

骆驼乳清蛋白对热应激所致大鼠肝氧化损伤的保护作用

本试验旨在研究骆驼乳清蛋白（CWP）对热应激（HS）大鼠肝损伤、氧化应激及肝功能的保护作用。选取6周龄SD大鼠36只,适应性饲养2周后随机分为6组：正常对照组饲喂基础日粮;CWP对照组添加400 mg·kg^-1的CWP;HS组除饲喂基础日粮外,每天进行2 h HS处理,连续8 d;3个CWP干预组分别于每次HS前灌服100、200和400 mg·kg^-1的CWP。试验结束后,取大鼠肝组织,HE染色观察肝组织病理学变化,同时检测肝功能生物标志物、氧化应激标志物水平及抗氧化酶活性。结果表明：CWP降低了HS大鼠血清谷草转氨酶（AST）和谷丙转氨酶（ALT）活性,400 mg·kg^-1的CWP干预效果最好（P<0.01）;400 mg·kg^-1CWP干预组有效降低了HS诱导的大鼠肝组织病理学改变; CWP降低了大鼠肝活性氧（ROS）及丙二醛（MDA）水平,增加了超氧化物歧化酶（SOD）、过氧化氢酶（CAT）、谷胱甘肽过氧化物酶（GSH-Px）活性及谷胱甘肽（GSH）水平,400 mg·kg^-1的CWP干预效果最好（P<0.01）。结果提示,CWP干预能够以剂量依赖性方式降低大鼠肝氧化应激,增加抗氧化系统防御能力,从而缓解HS所致大鼠肝损伤。     

Low potassium diets with different levels of calcium in comparison with different anionic diets fed to prepartum dairy cows: Effects on sorting behaviour,total tract digestibility,energy metabolism,oxidative status and hormonal response

This study investigated the effects of low potassium diets with different levels of Ca compared to two diets low in dietary cation–anion difference (DCAD) fed prepartum as a strategy to prevent hypocalcemia on sorting behaviour, total tract digestibility, oxidative status and energy and protein metabolism of transition cows. Forty-eight pregnant dairy cows were assigned to 4 treatment groups: Low Ca, low K (LCLK), High Ca, low K (HCLK), Supplementation with anionic mineral mixture (AMS) supplementation with SoyChlor (CAS). After parturition, all animals were fed a standard postpartum diet. Data were collected until 21 DIM. Prepartum urinary pH was significantly reduced by the low DCAD diets, while postpartum Ca homeostasis was affected by the HCLK ration. Feeding AMS induced sorting against particles <1.18 mm in favour of particles >19 mm prepartum. In contrast, cows fed CAS showed an increase in selective consumption of fine particles and sorted against longer particles similar to the HCLK and LCLK groups. Postpartum sorting activity was not affected by the dietary treatments. After calving, apparent digestibility of NDF was significantly reduced in the HCLK group. Prepartum, we observed effects on serum concentrations of non-esterified fatty acids were higher and insulin sensitivity was lower in the AMS group. Blood urea nitrogen (BUN) was decreased in cows fed the CAS ration. Postpartum, we found serum protein to be decreased with the low DCAD diets while BUN was decreased in the CAS group. The low DCAD rations increased prepartum serum malondialdehyde concentrations, while postpartum total antioxidant capacity was lower in the HCLK and the AMS group. From these data, we conclude that AMS decreased prepartum intake due to compromised palatability. Intermediate protein metabolism was affected by the low DCAD diets, while parameters of oxidative stress were probably affected by acid–base balance and Ca homeostasis.