ABSTRACT
Bacterial three-component dioxygenase systems consist of reductase and ferredoxin components which transfer electrons from NAD(P)H to a terminal oxygenase. In most cases, the oxygenase consists of two different subunits (alpha and beta). To assess the contributions of the alpha and beta subunits of the oxygenase to substrate specificity, hybrid dioxygenase enzymes were formed by coexpressing genes from two compatible plasmids in Escherichia coli. The activities of hybrid naphthalene and 2,4-dinitrotoluene dioxygenases containing four different beta subunits were tested with four substrates (indole, naphthalene, 2,4-dinitrotoluene, and 2-nitrotoluene). In the active hybrids, replacement of small subunits affected the rate of product formation but had no effect on the substrate range, regiospecificity, or enantiomeric purity of oxidation products with the substrates tested. These studies indicate that the small subunit of the oxygenase is essential for activity but does not play a major role in determining the specificity of these enzymes.
Subject(s)
Iron-Sulfur Proteins/metabolism , Multienzyme Complexes/metabolism , Oxygenases/metabolism , Burkholderia/enzymology , Dinitrobenzenes/metabolism , Dioxygenases , Escherichia coli/genetics , Indoles/metabolism , Iron-Sulfur Proteins/genetics , Multienzyme Complexes/genetics , Naphthalenes/metabolism , Naphthols/metabolism , Oxidation-Reduction , Oxygenases/genetics , Pseudomonas/enzymology , Recombinant Proteins/metabolism , Stereoisomerism , Substrate Specificity , Toluene/analogs & derivativesABSTRACT
Enucleated human eyes were banded with metal and silicone bands to produce reductions in their diameter of 2 mm and 4 mm. The ocular rigidity produced by each banding material at each diameter was measured in the pressure range of 10 mmHg to 40 mmHg. Metal bands produced mild reductions in ocular rigidity that were significantly (P less than 0.05 to 0.01) lower than the control ocular rigidities in some pressure ranges. Silicone bands produced large reductions in ocular rigidity that were significantly (P less than 0.01) lower than ocular rigidities observed in metal-banded or control conditions in all pressure ranges. The influence of the elastic silicone banding material on ocular rigidity was greater than the influence of altered shape and wall stress that occurred with metal banding.
