Chemically Defined Medium for the Accumulation of Intracellular Malate Dehydrogenase by Streptomyces aureofaciens
AUTOR(ES)
Laluce, Cecília
RESUMO
A chemically defined medium was developed for the production of intracellular malate dehydrogenases by Streptomyces aureofaciens NRRL-B 1286. The composition of the medium (per liter) was as follows: 50 g of starch, 4 g of ammonium sulfate, 7.32 g of l-aspartic acid, 13.8 g of MgSO4 · 7H2O, 1.7 g of K2HPO4, 0.01 g of ZnSO4 · 7H2O, 0.01 g of FeSO4 · 7H2O, 0.01 g of MnSO4 · H2O, and 0.005 g of CoSO4 · 7H2O. The pH of the medium was adjusted to 6.7 to 7.0 after sterilization. The activity of the intracellular malate dehydrogenases of the crude cell extract was greatest after 40 h of mycelium growth in a rotary shaker at 30°C. The best temperature for the enzyme reactions was approximately 35°C for NAD+ activity at pH 9.7 and 40°C for NADP+ -linked enzyme at pH 9.0. The NAD+ activity required Mg2+, and both activities were sensitive to SH-group reagents. The NADP+ -dependent activity remained completely stable, and the NAD+ -dependent activity decreased to a very low residual level after 30 min at 60°C.
ACESSO AO ARTIGO
http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=204024Documentos Relacionados
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