Addition of citrate to Acidithiobacillus ferrooxidans cultures enables precipitate-free growth at elevated pH and reduces ferric inhibition.
Acidithiobacillus ferrooxidans is an acidophilic chemolithoautotroph that is important in biomining and other biotechnological operations. The cells are able to oxidize inorganic iron, but the insolubility and product inhibition by Fe(3+) complicates characterization of these cultures. Here we explore the growth kinetics of A. ferrooxidans in iron-based medium in a pH range from 1.6 to 2.2. It was found that as the pH was increased from 1.6 to 2.0, the maintenance coefficient decreased while both the growth kinetics and maximum cell yield increased in the precipitate-free, low Fe(2+) concentration medium. In higher iron media a similar trend was observed at low pH, but the formation of precipitates at higher pH (2.0) hampered cell growth and lowered the specific growth rate and maximum cell yield. In order to eliminate ferric precipitates, chelating agents were introduced into the medium. Citric acid was found to be relatively non-toxic and did not appear to interfere with iron oxidation at a maximum concentration of 70 mM. Inclusion of citric acid prevented precipitation and A. ferrooxidans growth parameters resumed their trends as a function of pH. The addition of citrate also decreased the apparent substrate saturation constant (KS ) indicating a reduction in the competitive inhibition of growth by ferric ions. These results indicate that continuous cultures of A. ferrooxidans in the presence of citrate at elevated pH will enable enhanced cell yields and productivities. This will be critical as these cells are used in the development of new biotechnological applications such as electrofuel production.
Department of Chemical Engineering, Columbia University, 500W 120th Street, New York, New York, 10027.
Xiaozheng
X
Li
Roel
R
Mercado
Timothy
T
Kernan
Alan C
AC
West
Scott
S
Banta
D042763:Acidithiobacillus / Q000254:growth & development* / Q000378:metabolism; D018929:Cell Culture Techniques; D002614:Chelating Agents / Q000378:metabolism*; D011232:Chemical Precipitation; D019343:Citric Acid / Q000378:metabolism*; D003470:Culture Media / Q000378:metabolism; D005290:Ferric Compounds / Q000737:chemistry / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D007501:Iron / Q000378:metabolism*; D010084:Oxidation-Reduction
D002614:Chelating Agents; D003470:Culture Media; D005290:Ferric Compounds; D019343:Citric Acid; D007501:Iron
chemolithoautotrophy; iron chelation; iron-oxidizing bacteria; maintenance coefficient; product inhibition