海马水解蛋白的氨基酸组成与抗氧化能力的关系

为了探讨大海马蛋白质的高效利用方法,本试验将脱脂大海马粉经碱性蛋白酶和胰蛋白酶联合消解后,酶解液(PH)经超滤膜分离为5组分(PH-Ⅰ、PH-Ⅱ、PH-Ⅲ、PH-Ⅳ、PH-Ⅴ),结合各组分的氨基酸组成,回归分析了各组分的自由基清除能力与氨基酸组成间的相关关系。结果表明,酶解液经超滤膜分离的5组分中PH-Ⅴ (<2 500 Da)抗氧化能力较强,清除DPPH自由基能力为34.2%±0.1%,对超氧阴离子的清除能力为29.2%±0.1%,Fe³⁺还原力为0.28%±0.1%。海马酶解多肽各组成中疏水性氨基酸含量低于亲水性氨基酸,但疏水性氨基酸对自由基的清除能力起着决定性作用,极性氨基酸和非极性氨基酸可通过协同作用来增强海马多肽清除自由基的能力,且每个单位极性氨基酸的抗氧能力是非极性氨基酸的1.2倍,每个单位芳香族氨基酸的抗氧化能力是脂肪族氨基酸的2倍。本研究不仅制备了一类具有开发价值的抗氧化多肽,并且分析了多肽抗氧化能力与氨基酸组成的相关关系,为大海马的高值化利用提供了理论依据。     

PRRSV 2b蛋白与LGALS1相互作用的验证研究

旨在研究猪繁殖与呼吸综合症病毒(Porcine reproductive and respiratory syndrome virus,PRRSV)2b蛋白与宿主细胞蛋白之间的相互作用。本研究利用酵母回返试验、GST-pull down试验、免疫共沉淀试验3种方法对LGALS1与PRRSV 2b蛋白间是否相互作用进行验证。结果表明:在酵母回返试验中,含有LGALS1与2b基因的杂交酵母菌株在SD/-Trp-Leu-His-Ade/AbA/X-α-gal的平板上生长出蓝色菌落;在GST-pull down试验中,LGALS1与2b蛋白反应的样品经Western blot可检测到融合蛋白带;在免疫共沉淀试验中,LGALS1与2b蛋白反应的样品经HA单抗沉淀后,经Western blot可检测到相应蛋白带。该研究证明了LGALS1与PRRSV 2b蛋白间发生相互作用,为研究PRRSV感染途径和增殖过程提供了理论基础。     

Evaluation of three non‐genetically modified soybean cultivars as ingredients and a yeast‐based additive as a supplement in practical diets for Pacific white shrimp Litopenaeus vannamei

Two growth trials and a physiology assessment were conducted to evaluate three non‐genetically modified (GM) soybean cultivars as ingredients in practical diets for Pacific white shrimp, Litopenaeus vannamei. In addition, a commercially available fermented yeast product was evaluated as dietary supplement. For the growth trials (46 and 35 days, respectively, for trials 1 and 2), the basal diet was primarily composed of soybean meal (SBM), fishmeal (FM), whole wheat, corn protein concentrate, poultry meal (PM, pet food grade) and corn starch. Non‐GM cultivars were processed with novel methodologies to produce Navita? ingredients (N1, N2 and N3) which were incorporated at low (L) or high (H) levels into the experimental diets, in partial replacement of FM and full replacement of conventional SBM. The last two formulations incorporated the fermented yeast for a total of nine experimental diets (Table 1 ). Results from the growth trials indicate that shrimp fed diet 5 (HN2) exhibited significantly lower (p < .05) weight gain as compared to shrimp fed diets 1 and 2 (basal and LN1, respectively) in trial 1, as well as compared to animals fed diets 1, 2, 3 and 8 (basal, LN1, HN1 and basal + yeast) in trial 2. The feed conversion ratio significantly increased for shrimp fed diet 5, in contrast with shrimp fed diets 1, 2, 3 and 8 in trials 1 and 2, as well as compared to shrimp fed diets diet 6 (LN3) in trial 2. For the physiological assessment (stress and immune responses), only the effects of diets 1, 3, 8 and 9 (basal, HN1, basal + yeast and HN1 + yeast, respectively) were investigated. Granular cell counts were significantly higher for shrimp fed the yeast‐containing diets. Haemolymph glucose and haemolymph packed cell volume were significantly reduced for shrimp fed diets 3, 8 and 9. No significant differences were observed in total haemocyte counts, hyaline cells counts, semi‐granular cells counts, haemolymph protein, haemocyte phagocytic capacity and haemocyte respiratory burst activity. Results of this work indicate that selective soy breeding technology coupled with novel processing options has the potential to increase the nutritional value of conventional SBM for shrimp feeds. Trends on immune responses were more difficult to elucidate possibly due to the limited length of the feeding trial.     

Production of Antioxidative Maillard Reaction Products from Gelatin Hydrolysate of Unicorn Leatherjacket Skin

Antioxidative activities of Maillard reaction products (MRPs) derived from gelatin hydrolysate (GH) of unicorn leatherjacket skin as affected by types of saccharides, GH to saccharide ratio, incubation temperatures, relative humidity (RH), and times were investigated. MRPs prepared using mixture of GH and galactose at 2:1 (w/w) ratio showed the highest antioxidative activity. At different incubation temperatures (50–70°C) and RHs (55–75%), optimal condition for preparing antioxidative MRPs included heating GH and galactose (2:1) powder at 70°C and 55% RH for 36 h. Electrophoresis and Fourier transform infrared spectroscopy revealed the formation of high molecular weight polymers, and the advanced glycation took place in MRPs.     

陆地棉GhMYB9基因克隆及结合特性分析

MYB转录因子蛋白普遍存在于植物中,广泛参与植物的生长发育和代谢调控。为研究GhMYB9基因对棉纤维的调控机制,使用PCR方法,从棉花中克隆1个R2R3-MYB基因GhMYB9(Gen Bank登录号:AF336286.1);利用生物信息学方法分析其氨基酸序列;构建酵母表达载体。结果表明,该基因c DNA全长1 145 bp,开放阅读框长795 bp,编码264个氨基酸,预测其蛋白分子量约为29.624 k D,等电点为9.13。推测的氨基酸序列中含有2个高度保守的SANT结构域。GhMYB9基因组包含2个外显子和1个内含子。进化树分析表明,GhMYB9和Gh MYB聚在同一分支(HQ234875.1)。酵母单杂交试验结果表明,GhMYB9转录因子能够特异结合AC顺式作用元件。本研究结果为进一步阐明GhMYB9的生物学功能奠定了基础。     

利用酵母双杂交膜系统筛选与小麦蓝矮植原体免疫膜蛋白互作的异沙叶蝉蛋白

正小麦蓝矮病(Wheat blue dwarf,WBD)是我国西北麦区一种重要植原体病害,在我国西部地区危害严重。该病害由异沙叶蝉(Psammotettix alienus L.)专化性传播,介体传毒成为病害流行的重要中心环节[1]。本实验室前期通过免疫荧光标记研究发现WBD植原体免疫膜蛋白(immunodominant membrane protein, IMP)与介体异沙叶蝉肌动蛋白互作,说明IMP在植原体传播和致病过程中起关键作用。     

优化预处理及脱毒工艺以制备玉米芯半纤维素水解液

为了降低玉米芯半纤维素水解液中的乙酸、糠醛和酚类等有毒物质含量,使可利用糖类含量增加。本研究以玉米芯为原料制备水解液,对预处理及脱毒工艺进行优化,并利用高效液相色谱(HPLC)检测优化效果。结果表明,10~20目的玉米芯在10%氨水浸泡24 h后,可将乙酸和酚类物质的脱除率,分别提高至93.4%和32.3%,而糠醛的含量也大大降低。经过真空浓缩、CaO脱毒以及2%活性炭吸附后,使有毒物质含量再次降低,此时的水解液可完全用于菌株的发酵试验,达到了去除的目的。本文所得到的最佳水解液处理步骤,其结果符合菌株发酵用标准,说明水解液处理方式的不同,直接影响着水解液的制备。这为研究和利用木质纤维素等可再生资源提供了思路,也为发酵生产奠定了理论基础。     

Bacillus subtilis and yeast cell wall improve the intestinal health of broilers challenged by Clostridium perfringens

1. The objective was to investigate the effects of Bacillus subtilis, yeast cell wall (YCW) and their combination on intestinal health of broilers challenged by Clostridium perfringens over a 21-d period.

2. Using a 5 × 2 factorial arrangement of treatments, 800 1-d-old male Cobb 500 broilers were used to study the effects of feed additives (without additive or with zinc bacitracin, B. subtilis, YCW, and the combination of B. subtilis and YCW), pathogen challenge (without or with Clostridium perfringens challenge), and their interactive effects.

3. C. perfringens infection increased intestinal lesions scores, damaged intestinal histomorphology, increased serum endotoxin concentration, cytokine mRNA expression and intestinal population of C. perfringens and Escherichia coli and decreased ileal bifidobacteria numbers. The 4 additives decreased serum endotoxin. Zinc bacitracin tended to decrease cytokine mRNA expression and the intestinal number of C. perfringens and E. coli. B. subtilis, YCW and their combination increased cytokine mRNA expression. B. subtilis and YCW decreased the number of C. perfringens and E. coli in the ileum, and their combination decreased pathogens numbers in the ileum and caecum.

4. In conclusion, B. subtilis, YCW and their combination improved the intestinal health of NE-infected broilers, and could be potential alternatives to antibiotics.