A new method for pH determination in isoelectric focusing experiments

A rapid and reproducible method for the pH measurement in the effluent from density gradient electrofocusing is described. By this procedure, after preparative isoelectric focusing, the detection of protein zones and pH measurement can be accomplished simultaneously, by serially coupling a uv flow cell with a pH flow cell. This last one is connected to the recorder by a control unit, which allows the simultaneous printing of pH and uv absorption on the same chart.     

A new method to determine optimum temperature and activation energies for enzymatic reactions

A new method for determination of the optimum temperature and activation energies based on an idea of the average rate of enzymatic reaction has been developed. A mathematical model describing the effect of temperature on a dimensionless activity for enzyme deactivation following the first-order kinetics has been derived. The necessary condition for existence of the function extreme of the optimal temperature has been applied in the model. The developed method has been verified using the experimental data for inulinase from Kluyveromyces marxianus.     

A mathematical method for temperature compensation of pH measurements.

A continuous recording viscometer using oscillating capillary tubes has been developed. The basic principles, general design, and construction details of the viscosity sensing device are given. Two such devices, mechanically coupled, comprise a bridge viscometer with which two fluids can be compared directly. A bridge viscometer complete with the electronic amplifier circuits is described. The general design is readily adaptable to automatic systems.The instrument has a wide dynamic range. The fluid under test may be allowed to flow through the system. The capacity of each arm of the viscometer bridge is less than 100 μl.     

A new method for ethanol productivity determination

Summary A simple and easily applicable method for determination of the ethanol fermentation rate is described. The formulas to be used for the ethanol productivity calculation in the case of free and agar immobilized cells are given.     

A convenient pH-gradient method for the determination of the maximum and minimum pH for microbial growth

Previously existing methods for determining the pH limits for the growth of microorganisms have involved (1), the setting up of individual cultures, each having a specific pH; (2), the pH gradient plate technique devised by Sacks (1956) in which a continuous pH gradient is established in a Petri dish by means of a buffer system; and (3), the pH gradient plate technique of Zak (unpublished), in which a continuous pH gradient is established by means of an electric current. The discontinuous pH gradient technique described here provides a convenient method of determining the maximum and minimum pH at which a microorganism can grow. The technique can be used aerobically or anaerobically, and has a precision of about ± 0.1 pH unit. Data are given for several yeasts and forSerratia marcescens. In all cases, the organisms tested continued to metabolize at pH values beyond those representing the limits for growth, sometimes by as much as 0.5 pH unit. The results suggest that pH limits are unsuitable criteria in microbial classification.     

A new colorimetric method for determination of creatine phosphokinase

A new colorimetric method has been developed for the determination of creatine phosphokinase (CPK). This method is based on the reaction of creatine, formed enzymatically from creatine phosphate and ADP, with p-nitrophenylglyoxal (PNPG) under alkaline conditions to produce a colored product which absorbs maximally at 480 nm. At 25 degrees C the reaction was complete after 10 min in 0.1 M sodium pyrophosphate, pH 12, containing 0.15 M sodium ascorbate. The colored product was stable for at least 24 h and obeyed Beer's law in the range 0.005-0.05 mM creatine. The color reaction was used to determine the activity of CPK in serum and tissue extracts. The results obtained by this method agreed well with the results obtained by other available methods for CPK determination. However, the PNPG method was more rapid and more sensitive than other colorimetric methods and required a single chromogenic reagent.