Kinetic,thermodynamic parameters and in vitro digestion of tannase from Aspergillus tamarii URM 7115

Tannase is an enzyme used in various industries and produced by a large number of microorganisms. The aim of this study was to evaluate tannase production to determine the biochemical, kinetic, and thermodynamic properties and to simulate tannase in vitro digestion. The tannase-producing fungal strain was isolated from “jamun” leaves and identified as Aspergillus tamarii. Temperature at 26°C for 67?h was the best combination for maximum tannase activity (6.35-fold; initial activity in Plackett–Burman design—15.53?U/mL and average final activity in Doehlert design—98.68?U/mL). The crude extract of tannase was optimally active at 40°C, pH 5.5 and 6.5. Moreover, tannase was stimulated by Na⁺, Ca²⁺, Mg²⁺, and Mn²⁺. The half-life at 40°C lasted 247.55?min. The free energy of Gibbs, enthalpy, and entropy, at 40°C, was 81.47, 16.85, and ?0.21?kJ/mol?·?K, respectively. After total digestion, 123.95% of the original activity was retained. Results suggested that tannase from A. tamarii URM 7115 is an enzyme of interest for industrial applications, such as gallic acid production, additive for feed industry, and for beverage manufacturing, due to its catalytic and thermodynamic properties.     

Effect of select parameters on the properties of edible film from water-soluble fish proteins in surimi wash-water

Edible film from water-soluble fish proteins were developed by casting film solution on leveled trays and effects of pH (9.5, 10.0 and 10.5), heating temperature (60, 70 and 80 °C), and heating time (10, 20 and 30 min) of the film solution on various film properties were determined using Response Surface Methodology (RSM). The impact of pH and heating temperature of film solution was more significant, overall, on the film's properties than heating time. Contour plots of tensile strength and elongation at break was highest at pH of 10.0 at 70 °C (2.75-3.02 MPa) but low in elongation at break (6.35-9.16%), while water vapor permeability and oxygen permeability were at their lowest (58.55-65.96 g mm/m² d kPa and 351.33-624.18 cm³ μm/m² d kPa). There was a direct correlation between the films’ and proteins’ solubility on one hand, and heating temperature of film solution on the other, which reversed with change in pH of film solution. Film color was darker and more yellowish with increase in the pH of film solution.     

抗真菌蛋白Rs-AFP2基因定点突变及其对大肠杆菌中表达的影响

为了研究遗传密码子对表达调控的影响,利用PCR重叠延伸法,对萝卜抗真菌蛋白Rs-AFP2基因编码序列区的部分核苷酸进行沉默突变,构建突变体Rs-AFPm.序列分析表明,PCR产物全长240bp,有一个阅读框,编码的蛋白由29个氨基酸的信号肽和51个氨基酸的抗真菌蛋白组成.突变体与突变前的Rs-AFP2基因相比,在编码区第3号氨基酸Lys相差一个碱基(TTG→TTA),第5号氨基酸Gln相差一个碱基(CAG→CAA),第6号稀有密码子Arg相差两个碱基(CAG→CGA).重新合成引物,将切除信号肽的Rs-AFP2基因和Rs-AFPm基因与原核表达载体pET-21b(+)分别重组到大肠杆菌BL21菌株.IPTG诱导后,二者均得到了表达.软件分析显示,突变前pETAFPo表达产物占全菌蛋白的3%,突变后pETAFPm的表达产物占全菌蛋白含量的8%;表达蛋白主要以包涵体的形式存在,包涵体经超声波破碎后,蛋白质复性,抑菌结果表明,pETAFPm表达产物的抑菌半径大于pETAFP2表达产物的抑菌半径.这些都说明改造后的Rs-AFPm基因与Rs-AFP2基因相比,已有效地提高表达量.     

Effect of enzymatic hydrolysis of proteins on growth of Bifidobacterium bifidus in milk

The effect of enzymatic hydrolysis of proteins in milk using neutrase on the growth of the probiotic strain Bifidobacterium bifidus was evaluated by estimation of microbial growth, acidity, viscosity and flavour production. A significant increase in the growth of B bifidus was observed in neutrase‐hydrolysed milk. The setting time of bifidus‐cultured milk was advanced by about 12 h at 5% degree of hydrolysis. Enzymatic hydrolysis of proteins prior to cultivation also significantly increased the viscosity of the product. An approximately 60% increase in viscosity of the product was observed in neutrase‐hydrolysed milk. Production of steam‐volatile monocarbonyls as an indication of development of flavour was also higher in neutrase‐hydrolysed milk. The concentration of steam‐volatile monocarbonyls was 2.47 µmol per 100 ml in neutrase‐hydrolysed milk but only 1.84 µmol per 100 ml in control milk at the setting point of the curd. © 2002 Society of Chemical Industry     

给水管网计算中的数据转换方法

在给水管网计算中,通过建立环-管段矩阵、环-节点矩阵以及输入节点流量、节点坐标等少量数据,通过计算和数据转换,便可求得经济管径、管段设计流量、节点水压值、水厂供水量、管网造价、年费用折算值等。采用该方法进行管网计算,可以减少40%～70%的计算时间。     

Insertional re-activation of a chloramphenicol acetyltransferase misfolding mutant protein

The deletion of nine residues from the C-terminus of the bacterialchloramphenicol acetyltransferase (CAT) results in depositionof the mutant protein in cytoplasmic inclusion bodies and lossof chloramphenicol resistance in Escherichia coli. This foldingdefect is relieved by C-terminal fusion of the polypeptide withas few as two residues. Based on these observations, efficientpositive selection for the cloning of DNA fragments has beendemonstrated. The cloning vector encodes a C-terminally truncatedCAT protein. Restriction sites in front of the stop codon allowthe insertion of target DNA, resulting in the production ofproperly folded CAT fusion proteins and regained chloramphenicolresistance. The positive selection of recombinants is accomplishedby growth of transformants on chloramphenicol-containing agarplates. The method appears particularly convenient for the cloningof DNA fragments amplified by the PCR because minimal informationto restore CAT folding can be included in the primers. The cloningof random sequences shows that the folding defect can be relievedby fusion to a wide variety of peptides, providing great flexibilityto the positive selection system. This vector may also contributeto the determination of the role of the C-terminus in CAT folding.     

天然多糖类/蛋白质复合材料的研究进展

在参考了 31篇文献的基础上 ,详述了天然多糖类与蛋白质复合材料的研究进展     

Trypsin Inhibitor Activity In Vitro Digestibility and Sensory Quality of Meat-Like Yuba Products as Affected by Processing

ABSTRACT: :
Soybeans ( Glycine max ) were soaked and ground to obtain soymilk. The soymilk was cooked in an open tank and held at 85 to 90 deg;C. Yuba films were picked up in 20 min intervals and dried for 20 min. Yuba films were soaked in chicken-flavor solutions (25% and 35%), and baking soda (BS) solutions (0%, 1%, 2%, and 3% BS), and cooked at 100 °C for 30 min, 60 min, and 90 min. TIA decreased (p < 0.05) with the increase of heating time and BS concentration. In vitro protein digestibility (IVPD) decreased with heating time and BS concentration (p < 0.05). Sensory characteristics were affected by flavor concentration. By using 0% BS, 25% of the chicken flavor concentration, and a short heating time method, meat-like products with low TIA, high IVPD, and good sensory characteristics were obtained.     

Feasibility of fishmeal replacement by shrimp head silage protein hydrolysate in Nile tilapia (Oreochromis niloticus L) diets

This study provides information on the use of shrimp head silage protein hydrolysate (SPH) as an alternative protein source for tilapia feeding. Six diets (28% protein, 12% lipid) were prepared where fishmeal protein was replaced at levels of 0 (control), 10, 15, 20, 25 and 30% with the hydrolysate. The diets were supplied to Nile tilapia fry (338 mg initial weight) stocked in plastic recirculating 20 l tanks (10 animals per tank), with three replicates per treatment. After an 8 week experimental period, fish fed the diets containing 10 and 15% SPH showed significantly better performance in terms of final body weight, weight gain (%), mean daily weight gain (mg day^?1), specific growth ratio and feed conversion ratio than those fed the control diet (fishmeal as protein source) and higher‐SPH diets. It is concluded that shrimp head hydrolysate is a promising alternative protein source for tilapia feeding, improving growth ratio at dietary inclusion levels as high as 15%. In addition, the diets with added shrimp silage protein were well accepted by the fish, which avidly consumed the feed during the experiment. © 2002 Society of Chemical Industry     

从烟草悬浮细胞中分离液泡前体的方法研究

通过跟踪含有液泡前体特异标记物的组分, 利用细胞壁酶水解胞壁制造原生质体, 并用真针头挤压的方法裂解细胞, 用蔗糖密度梯度离心的方法分离液泡前体. 通过相差显微镜观察, 发现所分离的液泡前体保持完整的结构. 利用不同细胞器的标记物抗体进行免疫标记检测, 证明所纯化的液泡前体不含其他细胞器, 具有较高的纯度.