Effect of temperature on recombinant protein production using the Bm5/Bm5.NPV expression system

A series of experiments have been conducted using a recombinant baculovirus/insect cell expression system (Bm5/Bm5.NPV.CAT) to establish the optimum temperature for both cell growth and virus infection. Bm5 cell growth was found to be limited at temperatures below 22°C and ceased completely at temperatures above 34°C. In the range between 24 and 28°C, final cell densities always reached 96% of the highest achievable viable cell density. The shortest population doubling time was obtained at 28°C. Overall, a consistent increase in metabolism with increasing temperatures was observed. During the infection/viral replication phase, an increase in the temperature from 25 to 31°C resulted in a faster decrease in viable cell density and an earlier production of chloramphenicol acetyltransferase (CAT). Furthermore, protein yield at temperatures above 28°C was significantly reduced. Overall, the best temperature for the infection phase for the Bm5/Bm5.NPV expression system was found to be 25°C when the cells are cultured in serum free media.     

Predictive modelling of the 3-D structure of interleukin-13

Several atomic structures are now available for the family ofhelical cytokines, which includes growth hormone as well asmany of the interleukins. Using structural information fromfive members of this family, two alternative models of interleukin(IL)-13 are proposed. IL-13 has biological properties similarto those of IL-4 and, like the other interleukins, is a potentiallyimportant pharmaceutical target. The model of IL-13 is discussedand compared with the known interleukin structures.     

OPTIMUM-AIV: A planning and scheduling system for spacecraft AIV

This paper presents a project undertaken for the European Space Agency (ESA). The project is developing a knowledge based system for planning and scheduling of activities for spacecraft assembly, integration and verification (AIV). The system extends to the monitoring of plan execution and the plan repair phases.

The objectives of the contract are to develop an operational kernel of a planning, scheduling and plan repair tool, called OPTIMUM-AIV, and to provide facilities which will allow individual projects to customize the kernel to suit its specific needs. The kernel shall consist of a set of software functionalities for assistance in the initial specification of the AIV plan, in the verification and generation of valid plans and schedules for the AIV activities, and in interactive monitoring and execution problem recovery for the detailed AIV plans. Embedded in OPTIMUM-AIV are external interfaces which allow integration with alternative scheduling systems and project databases.

The current status of the OPTIMUM-AIV project, as of May 1991, is that the architectural design of the system has been agreed on by ESTEC/ESA and detailed design and implementation is now underway, expecting a final delivery in October of 1991.     

磺化竹红菌素对蛋白质荧光猝灭机理的研究

本文详细研究了磺化竹红菌素对多种蛋白质在溶液状态下的荧光猝灭过程。结果表明，磺化竹红菌素对多种蛋白质荧光猝灭服从Ｓｔｅｎ－Ｖｏｌｍｅｒ曲线。实验观察了温度、粘度、ｐＨ值和盐酸胍对荧光猝灭过程的影响。由于磺化竹红菌素是一两性分子，对于不同蛋白具有不同猝灭过程；磺化竹红菌素对蛋白质的荧光猝灭常数Ｋｑ在１０＾１３ｍｏｌ／Ｌ．ｓ＾－１左右，这说明，磺化竹红菌素是一种比其它蛋白质荧光猝灭剂更加有效的荧光猝灭     

大豆蛋白分离工艺研究

阐述了大豆蛋白质的分离原理,分析了运用“碱提酸沉”分离法提取大豆蛋白质过程中,豆粕的粉碎方式、浸提温度、浸提时间、浸提液的pH值以及离心工艺对提取效果的影响,为大豆蛋白／聚丙烯腈纤维的工业化生产提供了依据。     

Modulation of the enzymatic activity of papain by interdomain residues remote from the active site

The two main catalytic residues Cys25 and Hisl59 of the monomericcysteine protease papain are located on different walls of acleft formed by two domains. This topology suggests a possiblerelationship between relative domain organization and catalyticmechanism. The effect on enzymatic parameters of structuralmodifications at various locations of the twodomain interfaceof papain was examined by individual or double replacementsby Ala of pairs of interacting residues. Most modificationshad no effect on enzyme activity. However, the enzyme's substrateturnover (k_cat) decreased following simultaneous alterationof the two most conserved residues, forming an apolar contactlocated 15 Å away from the active site. The pH activityprofile of the double mutant was unchanged, indicating a conservedionization state of the active site thiolate-imidazolium ionpair. This state is strongly dependent on the distance separatingthe two residues, thus suggesting that the active site geometryhas not been significantly altered. Efficient enzymatic activityin papain requires more than a correct active site geometryand is influenced by domain packing properties in a region remotefrom the active site.     

Generation of a green fluorescent protein gene chromosomal insertion containing Escherichia coli strain for gene induction-based quantification of bioavailable lysine

The importance of lysine determination in feed materials is crucial for the feed industry because this amino acid can be limiting in many of the cereal materials used for animal feeds. The bacterial gene induction-based assay developed in this study aimed to measure lysine bioavailability in feeds as an alternative analytical method for animal assays. The advantages of a gene induction-based approach include rapid and quantitative estimation of many samples and results that relate a bacterial response to a biological response observed in animals. A whole-cell biosensor strain was constructed using a fluorescent E. coli strain that has an inducible fluorescent phenotype sensitive to extracellular lysine contents. A genetic fusion that links the promoter of cad operon with a green fluorescent protein encoding gene (gfp) was constructed, and a fluorescent assay was developed. A standard lysine curve (R² = 0.95) was generated and used for lysine bioavailability quantification of four feed ingredients (whole egg protein, blood-, soybean-, and meat and bone meal). Quantities as low as 50 μg/ml protein of digested samples were sufficient for analyses using the biosensor, except for meat and bone meal. Because of the low levels of free lysine in non-digested samples, fluorescence of these protein sources containing lower than 500 μg/ml protein was not detected (except for soybean meal). The results using enzymatically digested protein sources showed that the test strain emitted a fluorescent response that was proportional to the level of lysine present in the feed samples.     

蛋白质分子链的超声速孤立子

在Ｄａｖｙｄｏｖ简谐蛋白质分子链模型的基础上，进行合理的连续近似处理，得到了超声速和亚声速孤立子解，并求得孤立子能量．     

Whey Drying on Porous Carriers

Whey is an undurable product. treated very often as a waste which pollutes the natural environment. Whey which is a valuable source of protein, lactose, vitamins and mineral salts should be utilized completely. The present paper is a proposal of whey drying on porous carriers. It is proved experimentally that the proposed drying method guarantees good product quality.