Kinetics of protein release from yeast using enzymatic lysis for selective product recovery |
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Authors: | M J Torner B A Andrews J A Asenjo |
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Institution: | (1) Biochemical Engineering Laboratory, University of Reading, RG6 2AP Reading, U.K.;(2) Present address: IATA, (CSIC), C/.Jaime Roig, 11, 46010 Valencia, Spain |
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Abstract: | The kinetics of release of four intracellular enzymes from different yeast cell locations using the Differential Product Release
(DPR) method has been investigated. The method uses a combination of physical, chemical and biological agents such as lytic
enzymes, an osmotic support and a spheroplast stabilizer. Using the DPR technique a wall enzyme, invertase, was released with
a very high specific activity in the first step from a breadmaking strain ofS. cerevisiae. Maximum release could be obtained in this step when the incubation time was extended from 60 min to 100 min. Two cytosol
enzymes, α-D-glucosidase and alcohol dehydrogenase were released in the second step. Fumarase was released in the third step
almost instantaneously after disruption of the mitochondria which reduces considerably, by ca. 1 hour, the total incubation
time of DPR. This paper investigates the kinetics of enzyme release during the 3 steps of DPR. |
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