pH shift enhancement of Candida utilis pyruvate decarboxylase production.
Pyruvate decarboxylase (PDC) catalyses the synthesis of asymmetric carbinols, e.g., chiral precursors for pharmaceuticals such as ephedrine and pseudoephedrine. The production of PDC by Candida utilis in a minimal medium was improved by manipulating the pH during fermentation in a 5 L bioreactor. At an aeration rate of 0.1 vvm with a stirrer speed of 300 rpm at constant pH 6, a specific PDC activity of 141 U/g dry cell weight (DCW) was achieved (average of two fermentations +/-13%). By allowing the yeast to acidify the growth medium from pH 6 to 2.9, the final specific PDC activity increased by a factor of 2.7 to 385 U/g DCW (average from 4 fermentations +/-16%). The effect of this pH drift on PDC production was confirmed by another experiment with a manual shift of pH from 6 to 3 by addition of 5 M sulfuric acid. The final PDC activity was 392 U/g DCW (average from two fermentations +/-5%). However, experiments with constant pH of 6, 5, 4, or 3 resulted in average specific activities of only 102 to 141 U/g DCW, suggesting that a transitional pH change rather than the absolute pH value was responsible for the increased specific PDC activity.
Chen
Allen Kuan-Liang
AK
School of Biotechnology and Biomolecular Sciences, University of New South Wales, Sydney, Australia.
Breuer
Michael
M
Hauer
Bernhard
B
Rogers
Peter L
PL
Rosche
Bettina
B
D019149:Bioreactors / Q000382:microbiology*; D002175:Candida / Q000737:chemistry* / Q000201:enzymology* / Q000254:growth & development; D018929:Cell Culture Techniques / Q000379:methods*; D004789:Enzyme Activation; D004795:Enzyme Stability; D006863:Hydrogen-Ion Concentration; D011767:Pyruvate Decarboxylase / Q000096:biosynthesis* / Q000737:chemistry* / Q000302:isolation & purification
D011767:Pyruvate Decarboxylase