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In situ bubble fractionation strategies for separating individual proteins in a batch baker’s yeast fermentation process

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Abstract

Extracellular proteins produced by yeast have been observed to stratify in the extracellular fluid of a batch bioreactor, thus creating a vertical concentration gradient. We observed that, in the four different experiments conducted, each varied in their protein recovery characteristics. For example, sparging the system with gas accentuates the separation, though even in a nonsparged system, thein situ generation of minute carbon dioxide bubbles by yeast cells creates a protein gradient as the bubbles carry proteins upward. Based on these and other observations, we propose possible strategies for recovering the individual proteins from a system containing the four major proteins considered. A simple steady-state mathematical model, based on convective upward protein transport being balanced by downward protein diffusion, has been used to describe the behavior of each of these four extracellular proteins in the fermentation broth.

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Abbreviations

Cb :

protein concentration at the bottom of the fermentor

CL :

liquid concentration of an individual protein, mg/cm3

Cm :

protein concentration at the middle of the fermentor

Ct :

protein concentration at the top of the fermentor

D:

protein dispersion coefficient, cm2/s; k, 3QK/Dr (πR2), cm−1

kH:

separation efficiency factor

K:

proportionality (equilibrium) constant, F/Cl, cm

SR:

separation ratio, Ct/Cb

Q:

gas flow rate, cm3/s

X:

dimensionless protein concentration at position z

Xm :

dimensionless concentration at the fermentor midpoint (at z=H/2)

z:

column vertical position, cm (0<z<H), where z = 0 is the base position and z=H is the top of the fermentation broth Γ, bubble protein concentration, mg/cm2

r:

radius of bubble, cm

R:

radius of column, cm.

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DeSouza, A.H.G., Wilson, D.J. & Tanner, R.D. In situ bubble fractionation strategies for separating individual proteins in a batch baker’s yeast fermentation process. Appl Biochem Biotechnol 24, 565–578 (1990). https://doi.org/10.1007/BF02920279

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  • DOI: https://doi.org/10.1007/BF02920279

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