Preliminary study of the pyrolysis of steam classified municipal solid waste.

Steam classified municipal solid waste (MSW) has been studied for use as a combustion fuel, feedstock for composting, and cellulytic enzyme hydrolysis. A preliminary study has been conducted using a prototype plasma arc pyrolysis system (in cooperation with Plasma Energy Applied Technology Inc., Huntsville, AL) to convert the steam classified MSW into a pyrolysis gas and vitrified material. Using a feed rate of 50 lbs/h, 300 lbs of the material was pyrolysized. The major components of this pyrolysis gas were H(2), CO, and CO(2). A detailed presentation of the emission data along with details on the system used will be presented.

Investigation of the lignin-degrading activity of Serratia marcescens: biochemical screening and ultrastructural evidence.

Forty-one morphologically distinct bacterial isolates were developed from six lignin-containing environments. Each isolate was initially screened for potential lignin-degrading activity using relative growth on a lignocellulosic substrate and relative decolorization of a polymeric dye. Screened isolates were then tested for the ability to oxidize various lignin-related monomers, and the dimers anisoin and veratrylglycerol-beta-guaiacyl ether. Although most of the isolates oxidized the monomers, only two successfully oxidized the dimers. The dimer-degrading isolates were tested for extracellular activity against the beta-O-4 dimer veratryl-glycerol-beta-guaiacyl ether. No activity was detected for the isolates. Phanerochaete chrysosporium Burds used as a positive control demonstrated a high degree of activity in each assay. Extensive ultrastructural studies of lignocellulose alteration by the dimer-degrading isolates were conducted via light and transmission electron microscopy. These studies indicate that one of the isolates, identified as Serratia marcescens, is capable of degrading highly lignified secondary cell wall components. This activity is localized, apparently requiring direct contact between cells and substrate, which could be facilitated by an associated glycocalix. The results of the dimer degradation assays concur with the characterization of the responsible enzyme system as being membrane associated.

Reconstitution of the Escherichia coli pyruvate dehydrogenase complex.

The binding of pyruvate dehydrogenase and dihydrolipoyl dehydrogenase (flavoprotein) to dihydrolipoyl transacetylase, the core enzyme of the E. coli pyruvate dehydrogenase complex [EC 1.2.4.1:pyruvate:lipoate oxidoreductase (decaryboxylating and acceptor-acetylating)], has been studied using sedimentation equilibrium analysis and radioactive enzymes in conjunction with gel filtration chromatography. The results show that the transacetylase, which consists of 24 apparently identical polypeptide chains organized into a cube-like structure, has the potential to bind 24 pyruvate dehydrogenase dimers in the absence of flavoprotein and 24 flavoprotein dimers in the absence of pyruvate dehydrogenase. The results of reconstitution experiments, utilizing binding and activity measurements, indicate that the transacetylase can accommodate a total of only about 12 pyruvate dehydrogenase dimers and six flavoprotein dimers and that this stoichiometry, which is the same as that of the native pyruvate dehydrogenase complex, produces maximum activity. It appears that steric hindrance between the relatively bulky pyruvate dehydrogenase and flavoprotein molecules prevents the transacetylase from binding 24 molecules of each ligand. A structural model for the native and reconstituted pyruvate dehydrogenase complexes is proposed in which the 12 pyruvate dehydrogenase dimers are distributed symmetrically on the 12 edges of the transacetylase cube and the six flavoprotein dimers are distributed in the six faces of the cube.

A kinetic study of dihydrolipoyl transacetylase from bovine kidney.

The mammalian pyruvate dehydrogenase complex contains a core, consisting of dihydrolipoyl transacetylase, to which pyruvate dehydrogenase and dihydrolipoyl dehydrogenase are joined. This report describes studies on the kinetic mechanism of the transacetylase-catalyzed reaction between [1-14C]acetyl-CoA and dihydrolipoamide. This reaction appears to be a model of the physiological reaction, in which the acetyl group is transferred from the S-acetyldihydrolipoyl moiety, bound covalently to the transacetylase, to CoA. The model reaction is not affected by pyruvate dehydrogenase or dihydrolipoyl dehydrogenase, their substrates and products, or by removal of the covalently bound lipoyl moiety. These findings, together with the results of initial velocity, product inhibition, and dead-end inhibition studies, indicate that the model reaction and, apparently, the physiological reaction as well, proceeds via the Random Bi Bi (rapid equilibrium) mechanism. It appears that at the catalytic center of the transacetylase there are two adjacent sites, one that binds CoA and acetyl-CoA and another that binds dihydrolipoamide and S-acetyldihydrolipoamide (or the corresponding forms of the covalently bound lipoyl moiety.