Limited bacterial mineralization of fungal degradation intermediates from synthetic lignin.

The ability of selected bacterial strains and consortia to mineralize degradation intermediates produced by Phanerochaete chrysosporium from 14C-labeled synthetic lignins was studied. Three different molecular weight fractions of the intermediates were subjected to the action of the bacteria, which had been grown on a lignin-related dimeric compound. Two consortia isolated from wood being decayed naturally by a Ganoderma species of white rot fungus (the palo podrido system) mineralized 10 to 11% of the fraction with a molecular weight of approximately 500 but less than 4% of the higher- and lower-molecular-weight fractions. The consortia mineralized 5 to 9% of the original lignins. The ability of two pseudomonads isolated earlier from lignin-rich environments to mineralize the original lignins or fungus degradation products was much lower.

Biochemical and genetic studies of bacteria metabolizing lignin-related compounds.

The ability of bacterial strains to metabolize lignin model compounds was studied. Strains examined were non-filamentous bacterial isolates obtained from decaying wood and the actinomycete Streptomyces viridosporus T7A. Model compounds included dimers containing either the beta-1 (1,2-diarylethane) or the beta-O-4 (arylglycerol-beta-aryl ether) type of linkage. Pseudomonas fluorescens biovar I A1 proliferated on anisoin (4,4'-dimethoxybenzoin) accumulating anisic acid temporarily. Cleavage at the beta-1 bond was also observed with crude extracts prepared from the same strain. In turn, cleavage of the beta-O-4 linkage of veratrylglycerol-beta-guaiacyl ether was detected in cultures of Pseudomonas acidovorans D3. In this case, main degradation intermediates were beta-hydroxypropioveratrone, acetoveratrone and guaiacol. S. viridosporus T7A reduced the carbonyl group of some beta-1 dimers and did not modify the beta-O-4 model compounds tested. Attempts to ascribe a catabolic character to large molecular weight extrachromosomal DNA present in some strains were unsuccessful. Gene banks of P. fluorescens biovar I A1 and P. acidovorans D3 were prepared utilizing the broad host range cosmid pLAFR1 as vector.

DNA polymerases from the extremely thermophilic bacterium Thermus thermophilus HB-8.

Three DNA polymerase isoenzymes, which have been called A, B and C, were purified from Thermus thermophilus HB-8. These enzymes can be separated by chromatography (pH 7.5) on phosphocellulose and DNA-agarose. Their relative molecular masses, as determined by glycerol gradient centrifugation, fall in the range of 110000-120000. The three of them are devoid of exonuclease activity. Species A, B and C differ in their sensitivity towards N-ethylmaleimide (A greater than B greater than C) and urea (A greater than B = C) and also in their stability at high temperature (90 degrees C) (B greater than C greater than A). In addition, these enzymes can be distinguished utilizing various templates under different conditions. Thus, with activated DNA and Mg2+ as a cofactor, the highest incorporation is obtained at 50 degrees C with enzyme A and at 63 degrees C with enzymes B and C. If Mg2+ is replaced by Mn2+, the optimal temperatures remain unchanged, but enzyme A is stimulated twofold, while the activities of enzymes B and C decrease to one-half. On the other hand, with either poly(dA) X (dT)10 or poly(dA-dT) and Mg2+, enzyme A is inactive and enzyme C is severalfold more active than enzyme B. With the former synthetic template, optimal temperatures are 50 degrees C (enzyme C) and 40 degrees C (enzyme B), while with poly(dA-dT) they both work best at 63 degrees C. In turn, only enzyme C is able to utilize poly(rA) X (dT)10, although only with Mn2+ as a cofactor.