Culture conditions for the formation of dehydroacetic acid hydrolase (DHA hydrolase) and triacetic acid lactone hydrolase (TAL hydrolase) by
Pseudomonas sp. C-5-1, and some properties of their enzymes were investigated.
DHA in the culture medium was necessary for the enzyme formation and the optimum medium was determined as follows: DHA-Na, 0.2%; KH
2•PO
4, 0.1%; MgSO
4•7H
2O, 0.05%; NH
4NO
3, 0.1%; yeast extract, 0.1%. Both enzymes have been partially purified by DEAE-cellulose column chromatography and gel filtration on Sephadex G-75. Optimum pH and temperature for DHA hydrolase activity were 5.0 and 55_??_60°C, respectively, and the effect of metal ions such as Mg
2+, Mn
2+ Zn
2+, Cu
2+ and Hg
2+ on the activity was not determined.
On the other hand, optimum pH and temperature for TAL hydrolase activity were found to be 6.0_??_7.5 and 50°C, respectively. This enzyme was activated by the addition of Mn
2+, but was completely inhibited with EDTA, Cu
2+, Zn
2+ and Hg
2+.
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