Temperature control, enzyme

Fermentation. The term fermentation arose from the misconception that black tea production is a microbial process (73). The conversion of green leaf to black tea was recognized as an oxidative process initiated by tea—enzyme catalysis circa 1901 (74). The process, which starts at the onset of maceration, is allowed to continue under ambient conditions. Leaf temperature is maintained at less than 25—30°C as lower (15—25°C) temperatures improve flavor (75). Temperature control and air diffusion are faciUtated by distributing macerated leaf in layers 5—8 cm deep on the factory floor, but more often on racked trays in a fermentation room maintained at a high rh and at the lowest feasible temperature. Depending on the nature of the leaf, the maceration techniques, the ambient temperature, and the style of tea desired, the fermentation time can vary from 45 min to 3 h. More highly controlled systems depend on the timed conveyance of macerated leaf on mesh belts for forced-air circulation. If the system is enclosed, humidity and temperature control are improved (76). 

Temperature Control. While it was well known that enzyme catalysis is a direct function of temperature, little attention was paid to its control in kinetic enzyme assays until the pioneer work of Schneider and Willis (11). These workers showed that the temperature compartment of the Beckman DU spectrophotometer varied widely as a function of room temperature and of the number of times the cuvet compartment was opened. Thus, while most authors have assumed that they were conducting their assay at room temperature (i.e., a nominal 25 ) direct measurements showed that the cuvette temperature was closer to 32 C. Schneider and Willis suggested that thermospacers, hollow plates adjacent to each side of the cuvette compartment through which water at a constant temperature is circulated, be used in order to standardize clinical enzyme assay temperatures. 

The use of spectrophotometry to monitor enzyme-catalysed reactions (Table 8.6) is a very convenient and popular method owing to the simplicity of the technique and the precision that is possible. The technique lends itself readily not only to temperature control using water-jacketed or electrically heated cell holders but also to the measurement of initial velocities by continuous monitoring and recording techniques or by automated analysis systems. 

Production of the API begins with the selection of a synthetic route, as determined in the development program. Raw materials are added into a reaction vessel. These raw materials as reactants are heated or cooled in the reaction vessel (normal range is from -15 to 140 °C purpose-built vessels are needed for extreme reactions that require lower or higher temperature controls or pressurization of reaction processes). The chemical synthesis reactions are monitored and controlled via sensor probes (pH, temperature, and pressure) with in-process feedback controls for adjustments and alarms when necessary. Samples are withdrawn at dehned intervals for analysis to determine the reaction progress. Catalysts, including enzymes, may be added to speed up and direct the reaction along a certain pathway. 

Most ultrasonic experiments are carried out in temperature controlled systems to ensure that isothermal conditions are maintained. Even a small general increase in microbial temperature can influence both the active and passive transport systems of the cell membrane/wall and this in turn may lead to an increased uptake of compounds. If the temperature is not controlled then sonication could result in a large temperature increase which will lead to the denaturation (deactivation) of enzymes, proteins and other cellular components present within the microorganism [7]. 

Selected entries from Methods in Enzymology [vol, page(s)] Theory, 63, 340-352 measurement, 63, 365 cryosolvent [catalytic effect, 63, 344-346 choice, 63, 341-343 dielectric constant, 63, 354 electrolyte solubility, 63, 355, 356 enzyme stability, 63, 344 pH measurements, 63, 357, 358 preparation, 63, 358-361 viscosity effects, 63, 358] intermediate detection, 63, 349, 350 mixing techniques, 63, 361, 362 rapid reaction techniques, 63, 367-369 temperature control, 63, 363-367 temperature effect on catalysis, 63, 348, 349 temperature effect on enzyme structure, 63, 348. 

Temperature Ihe temperature in a bioreactor is an important parameter in any bioprocess, because all microorganisms and enzymes have an optimal temperature at which they function most efficiently. For example, optimal temperature for cell growth is 37 °C for Escherichia coli and 30 °C for Saccharomyces sp, respectively. Although there are many types of devices for temperature measurements, metal-resistance thermometers or thermistor thermometers are used most often for bioprocess instrumentation. The data of temperature is sufficiently reliable and mainly used for the temperature control of bioreactors and for the estimation of the heat generation in a large-scale aerobic fermentor such as in yeast production or in industrial beer fermentation. 

This problem led to the development of the composite we have mentioned earlier. Grafting the hydrophilic polyurethane onto the reticulated substratum while simultaneously immobilizing tlie enzyme allows us to maintain temperature control by passing cool air through the foam. We are not as concerned about cell structure that duty is handled by the reticulate. 

Ruffner, H. P., Hawker, J. S., and Hale, C. R. (1976). Temperature and enzymic control of malate metabolism in berries of Vitis vinifera. Phytochemistry 15,1877-1880. 

Proper pH and temperature control is critical during batch enzyme conversion processes, which usually last about 48 hours. In such processes, a number of enzyme tanks are filled sequentially from the converter at the adjusted temperature for treatment (140-150°F) and then dosed with the necessary enzymes. Progress of the reaction... 

Commercial dextrins are specifically the oligomers of starch. White dextrins, so called because of their visual appearance, are produced from a 30-40% suspension under the mildest possible hydrolysis conditions (79-120°C for 3-8 h in 0.2-2% H2S04 or HC1). Yellow dextrins and British gums are the partial hydrolysates at higher time-temperature integrals. Maltodextrins, dextrose equivalent20 5-19, derive from controlled enzyme or acid partial hydrolysis of gelatinized corn starch. The 20-24 dextrose equivalent hydrolysates tire com syrups (Appi, 1991). 

Environmental factors such as pH, ionic strength, and temperature affect enzyme activity. They must be controlled when making in vitro measurements of enzyme activity and heeded in vivo when abnormal conditions such as acidosis, alkalosis, and fevers may exist. 

An important case is the application of enzymes in laundry detergents. Market trends in the United States show that consumers prefer liquids to powder detergents by a ratio of 2 to 1. These products are stored with no temperature control on shelves in the presence of harsh surfactants, such as linear alkylbenzyl sulfonate (LAS) and require extraordinary measures for stabilization. LAS, by its nature as an effective cleaning agent, causes surfactant-induced unfolding in proteins. There are countless examples of the development of stabilization systems in the intellectual property space. A common theme is to reduce the water activity and to use borate/glycol stabilizers that bind to the active site of proteases. 

The effect of temperature on enzyme immunoassays should not be overlooked, and adequate thermo-static control is mandatory. This problem is not met with in other optical immunoassays which gives the latter some advantages. 

Some of the energy is used to form the cell structure. Reactions catalyzed by enzymes may be either endo- or exothermic depending on the particular stoichiometry. Because of the diluteness of the solutions normally handled, temperature control is achieved read-... 

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