Optimization of invertase immobilization by adsorption in ionic exchange resin for sucrose hydrolysis

This work presents as a main objective to study the immobilization process of yeast invertase by adsorption in the ion exchanging resin Duolite A-568 for invert sugar production. Initially, a kinetic study of the soluble form of the enzyme was carried out. At the sequence was studied the immobilization process of yeast invertase in the weakly exchanging anionic resin Duolite A-568. The influences of the pH, enzyme concentration and temperature in the enzyme immobilization were analyzed through a central composite design (CCD). The results indicated that the retention of the catalytic activity in immobilization was strongly dependent of these variables, being maximum in a pH value of 5.0, with an enzyme concentration of 12.5 g/L (1.875 g of protein per liter) and temperature of 30 °C. The simultaneous influence of pH and temperature on the free and immobilized invertase activity was also studied through a CCD.     

Ultrasound-Assisted Extraction of Water-Soluble Polysaccharides from the Fruit of Acanthopanaxbrachypus: Physicochemical,Structural and Functional Properties

The ultrasound-assisted extraction (UAE) parameters of total water-soluble polysaccharides (TABPs) from Acanthopanaxbrachypus fruit were optimized by Box-Behnken design (BBD) and response surface methodology (RSM). Physicochemical, structural, and functional properties of TABPs were investigated by chemical analysis, inductively coupled plasma optical emission spectrometry (ICP-OES), high performance liquid chromatography (HPLC), scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FT-IR), thermogravimetric analysis (TGA), water-holding capacity (WHC), oil-holding capacity (OHC), emulsion capacity (EC), emulsion stability (ES), as well as DPPH^. and ABTS^.+scavenging assays. The results showed that the maximal UAE-yield of TABPs was 3.81±0.18 % under the optimal conditions (ultrasonic power 325 W, extraction temperature 47 °C, extraction time 22 min). TABPs was rich in some beneficial element (Mg, K, Fe, Zn and Na) but little in harmful elements (Hg, Cd, As and Pb), and displayed rough surface with flake-like features and large dents, contained 93.89±0.08 % of total carbohydrate with more different monosaccharides including glucose, galactose, rhamnose, arabinose, mannose, xylose, and uronic acid in a molar ratio of 8.83 : 7.90 : 4.74 : 4.55 : 2.80 : 2.39 : 1.00, respectively. TABPs exhibited broad weight distribution (11.2–133.5 kDa), excellent thermal stability (>280 °C), WHC (0.61±0.08 g water/g sample) and OHC (4.53±0.12 g oil/g sample), as well as higher EC (43.75±1.23 %) and ES (38.32±1.50 %). Furthermore, TABPs also displayed remarkable scavenging activities on DPPH^. and ABTS^.+ in vitro. These findings provide a scientific basis for the applications of TABPs in functional additives for food, medicine, and cosmetics.     

传染性法氏囊病病毒弱毒株对Vero细胞的适应性研究

传染性法氏囊病病毒疫苗的传统生产工艺是利用鸡胚纤维细胞增殖ＩＢＤＶ,但易被外源病原污染,且生产成本很高,借鉴国内外流行性乙型脑炎疫苗生产的经验,使用Ｖｅｒｏ细胞增殖ＩＢＤＶ弱毒株,从病毒高峰出现时间、滴度峰值的高低、对细胞代谢的影响等方面研究了ＩＢＤＶ对Ｖｅｒｏ细胞的适应性,敏感性以及适应后的种毒的长期保存过程中发现病毒滴度有下降趋势,试用了不同试剂对病毒进行保护。     

Anionic Antimicrobial and Anticancer Peptides from Plants

Anionic antimicrobial peptides (AAMPs) have been identified in a wide variety of plant species with net charges that range between ?1 and ?7 and structures that include: extended conformations, α-helical architecture and cysteine stabilized scaffolds. These peptides commonly exist as multiple isoforms within a given plant and have a range of biological activities including the ability to kill cancer cells as well as phytopathogenic bacteria, fungi, pests, molluscs, and other predatory species. In general, the killing mechanisms underpinning these activities are poorly understood although they appear to involve attack on intracellular targets such as DNA along with compromise of cell envelope integrity through lysis of the cell wall via chitin-binding and/or permeabilisation of the plasma membrane via lipid interaction. It is now becoming clear that AAMPs participate in the innate immune response of plants and make a major contribution to the arsenal of defence toxins produced by these organisms to compensate for their lack of some defence mechanisms possessed by mammals, such as mobility and a somatic adaptive immune system. Based on their biological properties, a number of potential uses for plant AAMPs have been suggested, including therapeutically useful anticancer agents and novel antimicrobial compounds, which could be utilized in a variety of scenarios, ranging from the protection of crops to the disinfection of hospital environments.     

Nutritional impacts of dietary oregano and Enviva essential oils on the performance,gut microbiota and blood biochemicals of growing ducks

Nowadays, there is much legislation in the world devoted to restrict the use of synthetic antibiotics in the poultry industry, which could reduce performance rate and production profits. Various phyto-biotic growth promoters have been proposed to serve as antibiotic alternatives with emphasis on plant extracts and essential oils. This study was conducted to assess the impacts of using the oregano essential oil (OEO) (comprised of 5% thymol and 65% carvacrol) and Enviva essential oil (EEO) (4.5% cinnamaldehyde and 13.5% thymol) as phytobiotic feed additives (PFA) on growth performance, cecal microbiota and serum biochemicals of growing ducks. In total, 800 11-day-old ducklings, housed in 20 floor pens, were allotted randomly into five dietary treatments: (i) A basal diet (BD) (control), (ii.) BD+50 mg EEO/kg, (iii.) BD+100 mg EEO/kg, (iv.) BD+150 mg OEO/kg and (v.) BD+300 mg OEO/kg diet. The growth performance traits were studied between 11 and 42 days of age. At the experiment end, 40 ducks were slaughtered (eight/ treatment) and cecal digesta and blood samples were collected to estimate the cecal bacterial populations and serum blood biochemicals. The results indicated that the tested levels of OEO and EEO did not display any significant effect (P>0.05) on the duck’s final BW, BW gain, growth rate, feed intake, feed conversion ratio or survivability rate. Besides, the different levels of EEO and OEO decreased the cecal populations of Coliforms (P<0.01), total aerobes (P<0.01) and lactose-negative Enterobacteria (P<0.05) in comparison with those of the control group. Finally, the tested EEO and OEO levels did not show any significant effect on the serum variables; in terms of total protein, albumin, globulin, total cholesterol, triglycerides, alanine aminotransferase and aspartate aminotransferase. In conclusion, the antimicrobial effect of the OEO and EEO against the cecal microbiota has been proven, while they did not display significant effects on the growth performance or blood variables of growing ducks.     

桉叶饲料添加剂的研制和应用

桉树叶及其提取物是近期开发研究的一种新型饲料添加刺。在采用柠檬桉和窿缘桉等树叶作原料,单罐浸提,选择到最适工艺条件为浸提时间60分钟,溶剂浓度60％,料液比1:6—7,浸提次数2次,可取得较好的浸提效果。饲养禽畜试验表明,桉叶饲料添加剂能使猪增重13％左右,饲料报酬提高5—10％,蛋鸡提高产蛋2—3％,死亡率降低1％。肉鸡增重提高3％,饲科报酬提高4—8％,有显著的经济效益。     

黑蒴定芽诱发增殖的初步研究

黑蒴(Melasma arvense)为云南省民族民间珍稀药材,现已成为云南天然药物开发的热点药材种类.但目前在云南已濒临灭绝,急需对其进行资源保护和人工繁育.该研究在筛选黑蒴不同器官培养的基础上,以茎枝为外植体,依次考察了不同基本培养基、不同种类细胞分裂素和质量浓度、不同天然有机添加物对定芽(顶芽和侧芽)萌发增殖及生长的影响.结果表明:以黑蒴带节茎枝为外植体,将其剪成2~3 cm带节嫩茎枝,经严格的外植物体消毒程序处理后,接种到1/2MS+6-BA 0.5 mg·L-1+Pt 50 g·L-1+Bn 80 g·L-1+25 g·L-1蔗糖+7 g·L-1琼脂,pH值5.8的培养基上,接种后放置在光照强度1500~2000 lx,光照10 h·d-1,温度22℃下培养,接种后8~10 d开始萌发,40 d单茎平均增殖数达8.1,主茎40 d生长长度74.2 mm,显示出极好的诱发增殖生长效果.该研究结果初步建立了黑蒴嫰茎枝诱发定芽及增殖培养体系,为云南珍稀民族药黑蒴的资源保护、人工栽培和可持续利用提供了科学依据.     

Activities of anionic and cationic trypsins in the temperature range from 5 to 37 degrees C. Mutant anionic trypsins as a model of cold-adapted (psychrophilic) enzymes

Temperature dependences of kinetic constants (k _cat and K _m) were studied for enzymatic hydrolysis of N-succinyl-L-alanyl-L-alanyl-L-prolyl-L-arginine-p-nitroanilide and N-succinyl-L-alanyl-L-alanyl-L-prolyl-L-lysine-p-nitroanilide by bovine cationic and rat anionic (wild-type and mutant) trypsins. The findings were compared with the corresponding literature data for hydrolysis of N-benzoyl-DL-arginine-p-nitroanilide by bovine cationic trypsin and natural trypsins of coldadapted fishes. The anionic and cationic trypsins were found to differ in organization of the S₁ -substrate-binding pocket. The difference in the binding of lysine and arginine residues to this site (S₁) was also displayed by opposite temperature dependences of hydrolysis constants for the corresponding substrates by the anionic and cationic trypsins. The data suggest that the effect of any factor on the binding of substrates (the K _m value) to the anionic and cationic trypsins and on the catalytic activity k _cat should be compared only with the corresponding data for the natural enzyme of the same type. Mutants of rat anionic trypsin at residues K188 or Y228 were prepared by site-directed mutagenesis as approximate models of natural psychrophilic trypsins. Substitution of the charged lysine residue in position 188 by hydrophobic phenylalanine residue shifted the pH optimum of the resulting mutant trypsin K188F from 8.0 to 9.0-10.0, similarly to the case of some natural psychrophilic trypsins, and also 1.5-fold increased its catalytic activity at low temperatures as compared to the wild-type enzyme.     

Comparative Evaluation of α-Amylase Refolding Through Two Different Artificial Chaperone Systems

Two different artificial chaperone systems were evaluated in this work using either detergents or CDs as the stripping agents. Upon dilution of urea-denatured α-amylase to a non-denaturing urea concentration in the presence of the capturing agent, complexes of the detergent and non-native protein molecules are formed and thereby the formation of protein aggregates is prevented. The so-called captured protein is unable to refold from the detergent-protein complex states unless a stripping agent is used to remove the detergent molecules. Our results by fluorescence, UV, turbidity measurement, circular dichroism, surface tension and activity assay indicated that the extent of refolding assistance was different due to different inter- and intra- molecular interactions in the two different systems. However, the high activity recovery in the presence of detergents, as the stripping agent, suggests that they can constitute suitable replacement for the more expensive and common stripping agent of cyclodextrins.