Evaluation of column flotation in the downstream processing of fermentation products: recovery of a genetically engineered alpha-amylase.
Flotation is a simple, inexpensive, and versatile unit operation with a largely unexplored potential in biotechnology. There is a general lack of research concerning biotechnological applications in this area, especially in the recovery of fermentation products. Moreover, the few reports in the literature do not consider the modern concept of column flotation as practiced in the mineral industry. We report herein the application of column flotation for the recovery of a Bacillus stearothermophilus alpha-amylase expressed in Escherichia coli by the use of a food-grade polymer, (hydroxypropyl)methylcellulose (HPMC), and ammonium sulfate. First, the enzyme was removed from the liquid phase by partition to a salted-out HPMC phase. The enzyme-containing polymer flocs were then floated from the liquid. Recovery of active enzyme was as high as 90%, with throughput as high as 94 m3/(h.m2). The floatability of the enzyme from a periplasmic extract was higher than extracellular enzyme in the broth due to the presence of depressors of molecular weight lower than 10,000 in the broth.
Miranda
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Department of Agricultural Engineering, Michigan State University, East Lansing 48824.
Berglund
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D000645:Ammonium Sulfate; D001426:Bacterial Proteins / Q000235:genetics / Q000302:isolation & purification*; D001709:Biotechnology / Q000379:methods*; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D005069:Evaluation Studies as Topic; D005285:Fermentation; D005818:Genetic Engineering / Q000379:methods; D001411:Geobacillus stearothermophilus / Q000201:enzymology / Q000235:genetics; D011108:Polymers; D000516:alpha-Amylases / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification*
D001426:Bacterial Proteins; D011108:Polymers; D000516:alpha-Amylases; D000645:Ammonium Sulfate