Expression of a recombinant crustacean hyperglycemic hormone of the kuruma prawn Penaeus japonicus in methylotrophic yeast Pichia pastoris

ABSTRACT:   Until now, six crustacean hyperglycemic hormones (CHH) designated Pej-SGP-I, -II, -III, -V, -VI and -VII have been characterized in the kuruma prawn Penaeus japonicus . All CHH consist of 72 amino acid residues and have an amidated carboxyl (C)-terminus. In the present study, we expressed Pej-SGP-III in methylotrophic yeast Pichia pastoris in order to obtain a large quantity of recombinant CHH possessing biological activity. A cDNA encoding Pej-SGP-III that had been previously cloned was processed by polymerase chain reaction (PCR) and the resulting product was ligated into an expression vector. Pichia pastoris was transformed with this vector after which a recombinant Pej-SGP-III was expressed having an additional amino acid residue (glycine) at the C-terminus (rPej-SGP-III-Gly), a form considered to be a putative precursor of this hormone. rPej-SGP-III-Gly secreted into the culture medium was purified by reversed-phase HPLC, and amidated using a peptidylglycine alpha-amidating enzyme. The amidated rPej-SGP-III (rPej-SGP-III) showed hyperglycemic activity in in vivo bioassay almost comparable to that of the natural Pej-SGP-III. rPej-SGP-III thus obtained will be a useful tool not only for its physiological study but also for the determination of its 3-D structure.     

A review of antimicrobial peptides: defensins and related cationic peptides

Cationic antimicrobial peptides are present throughout the plant and animal kingdoms and bear striking structural and functional similarities across species lines. They provide primitive, nonspecific means of combating a variety of bacteria, fungi, enveloped viruses, and protozoa. Some are also cytotoxic against host cells, including neoplastic cells. Cationic antimicrobial peptides may play various roles in inflammation and tissue repair. Antimicrobial peptides are found in epithelial tissues regularly exposed to microbial attack as well as in cells whose primary function is defense against potential pathogens. They constitute an important part of the nonoxidative antimicrobial arsenal of leukocytes. They are preformed and/or readily synthesized when the cells are stimulated by exposure to pathogens. They exert their effects directly by inserting into membranes of target cells and forming ion channels which increase membrane permeability; however, antimicrobial peptides can also act as opsonins to facilitate phagocytosis. Resistance to defensins is a virulence factor for organisms such as Salmonella sp. The study of cationic antimicrobial peptides is increasing our understanding of innate immunity, inflammation, and the pathogenesis of genetic diseases such as specific granule disease in humans. Therapeutic applications of antimicrobial peptides are currently under investigation.     

饲粮中添加抗菌肽对肉鸭增重及血清尿素氮、总蛋白水平的影响

240只1日龄肉鸭随机成分4组,每组设3个重复,对照组用基础日粮,试验组在基础日粮上每千克分别添加1、2、3mL抗菌肽制剂。结果显示:1)2、3mL/kg添加量组增重效果显著(P<0.05),3mL/kg添加量组最佳;净肉率升高,达到显著水平(P<0.05),其中主要是胸肌率上升;腹脂率降低但差异不显著(P>0.05);料重比无显著变化(P>0.05)。2)血清总蛋白浓度差异不显著(P>0.05),但尿素氮浓度下降,2、3mL/kg添加量组达显著水平(P<0.05),3mL/kg添加量组最低。     

Biosynthesis of Depsipeptide Mycotoxins in Fusarium

The cyclic hexadepsipeptide enniatin is known as a phytopathogenic compound from Fusaria causing necrosis and wilt. The molecule consists of three alternating residues each of a branched chain amino acid and D-hydroxyisovaleric acid (D-Hiv). Enniatins are synthesized by a 347kDa multienzyme (enniatin synthetase) via a thiol template mechanism. The corresponding gene esyn1 has an open reading frame of 9393 nucleotides and harbours two modules, one responsible for D-hydroxy acid activation and one for L-amino acid activation with an integrated N-methyltransferase domain. Such methyltransferases build an homologous group among N-methyl peptide synthetases. Enniatins are synthesized by step-wise condensation of dipeptidol building blocks in an iterative manner resembling fatty acid synthesis. A key enzyme in enniatin biosynthesis is the NADPH-dependent D-2-hydroxyisovalerate dehydrogenase, that supplies enniatin synthetase with D-Hiv. Enniatins contribute to the wilt toxic character of Fusaria. Virulence was significantly reduced in F. avenaceum after disruption of the esyn1 gene.     

法氏囊活性肽BP7调节鸡未成熟B细胞的基因表达谱及功能分析

试验旨在探索法氏囊活性肽BP7调节鸡未成熟B细胞的分子基础.利用BP7刺激禽前B淋巴细胞DT40细胞,采用荧光定量PCR(qPCR)检测IgM的mRNA水平,并采用基因芯片分析基因表达谱及其生物学功能.结果 显示,BP7刺激的DT40细胞产生IgM的mRNA水平明显升高.基因芯片分析发现,BP7处理的DT40细胞中共有...     

黄芪多糖的超声波-闪式协同提取工艺优化及抗氧化活性研究

以黄芪为原料,采用超声波-闪式协同提取黄芪多糖。以黄芪多糖得率为评价指标,采用单因素试验和正交试验,确定最佳提取工艺。结果表明,超声波提取最佳工艺条件为：料液比1∶30（g/mL）,超声温度70 ℃,超声时间40 min,超声波功率500 W,在此条件下,黄芪多糖得率为7.11%;在对超声波提取工艺优化的基础上,协同闪式提取,最佳闪式提取工艺参数为：提取次数3 次,闪提时间100 s,提取电压160 V,在此条件下,黄芪多糖得率为11.03%;体外抗氧化活性试验结果表明：当黄芪多糖质量浓度为1.0 mg/mL 时,其对1,1-二苯基-2-三硝基苯肼（1,1-diphenyl-2-picrylhydrazyl,DPPH）自由基和2,2-联氨-双-3-乙基苯并噻唑啉-6-磺酸（2,2’-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid),ABTS）阳离子自由基的清除率分别为66.27%和58.27%。本研究获得了黄芪多糖的最佳提取方法,得到的黄芪多糖具有较强的抗氧化作用。该方法可为黄芪多糖的提取及开发应用提供参考。     

甘肃沙枣多糖提取工艺研究

采用热水浸提、醇沉法提取沙枣多糖,对沙枣多糖提取工艺进行了研究。选择料液比、浸提温度和浸提时间作为单因素进行梯度实验,确定其提取备件范围,再通过正交试验进一步确定沙枣多糖最佳提取工艺条件。结果表明,沙枣多糖的最佳提取工艺为：浸提温度为85℃、固液比为1：60、浸提时间为2h,在此条件下提取得甘肃沙枣多糖含量为4．98％。     

抗菌肽对抗生素耐药菌株的抑菌活性研究

[目的]研究抗菌肽对抗生素耐药菌株的抑菌活性。[方法]利用抗性平板划线法从腹泻病牛血便中筛选分离出1株耐药菌,通过16S rDNA序列进行鉴定,采用琼脂孔穴扩散法通过梯度盐酸壮观霉素（spectinomycin,Spe⁺）、氨苄青霉素（ampicillin,Amp⁺）、硫酸卡那霉素（kanamycin,Kan⁺）和氯霉素（chloramphenicol,Cm⁺）试验确定该菌药敏特性,并利用1种抗菌肽制剂对该菌株进行药敏试验。[结果]经BLAST比对分析该菌16S rDNA序列,鉴定该耐药菌为科氏葡萄球菌（Staphylococcus cohnii）,此菌对Amp⁺敏感,但对试验中其他抗生素均有耐药性,各梯度抗菌肽对该耐药菌均具有明显的抑菌活性。[结论]抗菌肽能有效抑制耐药科氏葡萄球菌的生长,有望在畜牧生产中代替抗生素使用。     

酵母细胞壁多糖在动物生产中的应用

酵母细胞壁多糖(YCP)作为一种天然绿色的添加剂,具有促进动物生长发育、提高动物生产性能、保护动物肠道健康和增强动物免疫力等诸多功能,可以有效替代抗生素在动物饲料中广泛使用.本文就YCP的组成结构以及在动物生产中的应用进行综述,为其进一步的研究和利用提供理论依据.     

茗荷水溶性多糖提取及抗氧化性能测定

采用单因素试验和厶(34)正交试验设计,研究了不同的料液比、提取时间和提取温度对茗荷多糖提取率的影响.并采用邻苯三酚自氧化法测定茗荷多糖对超氧阴离子自由基(O2-.)清除活性,Fenton体系测定茗荷多糖对羟基自由基(·OH)的清除作用.茗荷多糖提取的最佳工艺条件为以1:30的科液比,在90℃水浴条件下.提取时间为4h;茗荷多糖对超氧阴离子自由基和羟基自由基有较强的清除作用.