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Biomedical subjects

T K Kirk

Publications and source records attributed to T K Kirk.

At least 55 records · Page 3Linked to original sources

Relationship Between Lignin Degradation and Production of Reduced Oxygen Species by Phanerochaete chrysosporium.

The relationship between the production of reduced oxygen species, hydrogen peroxide (H(2)O(2)), superoxide (O(2)), and hydroxyl radical (.OH), and the oxidation of synthetic lignin to CO(2) was studied in whole cultures of the white-rot fungus Phanerochaete chrysosporium Burds. The kinetics of the synthesis of H(2)O(2) coincided with the appearance of the ligninolytic system; also, H(2)O(2) production was markedly enhanced by growth under 100% O(2), mimicking the increase in ligninolytic activity characteristic of cultures grown under elevated oxygen tension. Lignin degradation by whole cultures was inhibited by a specific H(2)O(2) scavenger, catalase, implying a role for H(2)O(2) in the degradative process. Superoxide dismutase also inhibited lignin degradation, suggesting that O(2) is also involved in the breakdown of lignin. The production of .OH was assayed in whole cultures by a benzoate decarboxylation assay. Neither the kinetics of .OH synthesis nor the final activity of its producing system obtained under 100% O(2) correlated with that of the lignin-degrading system. However, lignin degradation was inhibited by compounds which react with .OH. It is concluded that H(2)O(2), and perhaps O(2), are involved in lignin degradation; because these species are relatively unreactive per se, their role must be indirect. Conclusions about a role for .OH in ligninolysis could not be reached.

Journal Article↗

Nutritional Regulation of Lignin Degradation by Phanerochaete chrysosporium.

Previous studies have shown that a lignin-degrading system appears in cultures of the white rot fungus Phanerochaete chrysosporium in response to nitrogen starvation, apparently as part of secondary metabolism. We examined the influence of limiting carbohydrate, sulfur, or phosphorus and the effect of varying the concentrations of four trace metals, Ca, and Mg. Limitation of carbohydrate or sulfur but not limitation of phosphorus triggered ligninolytic activity. When only carbohydrate was limiting, supplementary carbohydrate caused a transient repression of activity. In carbohydrate-limited cultures, ligninolytic activity appeared when the supplied carbohydrate was depleted, and this activity was associated with a decrease in mycelial dry weight. The amount of lignin degraded depended on the amount of carbohydrate provided, which determined the amount of mycelium produced during primary growth. Carbohydrate-limited cultures synthesized only small amounts of the secondary metabolite veratryl alcohol compared with nitrogen-limited cultures. l-Glutamate sharply repressed ligninolytic activity in carbohydrate-starved cultures, but NH(4) did not. Ligninolytic activity was also triggered in cultures supplied with 37 muM sulfur as the only limiting nutrient. The balance of trace metals, Mg, and Ca was important for lignin degradation.

Journal Article↗

Ligninolytic enzyme system of Phanaerochaete chrysosporium: synthesized in the absence of lignin in response to nitrogen starvation.

The relationship between growth, nutrient nitrogen assimilation, and the appearance of ligninolytic activity was examined in stationary batch cultures of the wood-destroying hymenomycete Phanerochaete chrysosporium Burds. grown under conditions optimized for lignin metabolism. A reproducible sequence of events followed inoculation: 0 to 24 h, germination, linear growth, and depletion of nutrient nitrogen; 24 to 48 h, cessation of linear growth and derepression of ammonium permease activity (demonstrating nitrogen starvation); 72 to 96 h, appearance of ligninolytic activity (synthetic 14C-lignin leads to 14CO2). Experiments with cycloheximide demonstrated that appearance of ligninolytic activity occurs irrespective of the presence of lignin; lignin did not induce additional activity. Addition of NH4+ to cultures immediately prior to the time of appearance of the ligninolytic system delayed its appearance, suggesting that the NH4+ led to interference with synthesis of the enzyme system. Addition of NH4+ to ligninolytic cultures resulted in an eventual, temporary decrease in ligninolytic activity. The results suggest that all or essential protein components of the ligninolytic enzyme system are synthesized as part of a series of physiological ("secondary metabolic") events that are initiated by nutrient nitrogen starvation.

Basidiomycota↗

Microbial decomposition of synthetic C-labeled lignins in nature: lignin biodegradation in a variety of natural materials.

Lignin biodegradation in a variety of natural materials was examined using specifically labeled synthetic C-lignins. Natural materials included soils, sediments, silage, steer bedding, and rumen contents. Both aerobic and anaerobic incubations were used. No C-labeled lignin biodegradation to labeled gaseous products under anaerobic conditions was observed. Aerobic C-labeled lignin mineralization varied with respect to type of natural material used, site, soil type and horizon, and temperature. The greatest observed degradation occurred in a soil from Yellowstone National Park and amounted to over 42% conversion of total radioactivity to CO(2) during 78 days of incubation. Amounts of C-labeled lignin mineralization in Wisconsin soils and sediments were significantly correlated with organic carbon, organic nitrogen, nitrate nitrogen, exchangeable calcium, and exchangeable potassium.

Journal Article↗

Preparation and microbial decomposition of synthetic [14C]ligins.

A definitive assay for microbiological and biochemical research on the biodegradation of lignin was developed using radioactive synthetic lignins specifically labeled in the side chains, aromatic rings or in the methoxyl groups. The [14C]lignins were prepared by oxidative polymerization with peroxidase and H2O2 Of specifically labeled coniferyl alcohol (4-hydroxy-3-methyoxycinnamyl alcohol). The synthetic polymers were shown by spectroscopic and chemical methods to contain the same intermonomer linkages found in natural lignins. Incubation of the [14C]lignins with known lignin-degrading fungi and with a forest soil resulted in 14CO2 evolution.

Agaricales↗

Oxygenation of 4-alkoxyl groups in alkoxybenzoic acids by Polyporus dichrous.

The degradation of several alkyl ethers of vanillic acid, of 3-ethoxy-4-hydroxybenzoic acid, and of syringic acid, by the lignin-decomposing fungus Polyporus dichrous included (i) 4-dealkylation (e.g., 3-ethoxy-4-isopropoxybenzoic acid was in part dealkylated to 3-ethoxy-4-hydroxybenzoic acid), (ii) hydroxylation of the 4-alkoxyl groups (e.g., 3-ethoxy-4-isopropoxybenzoic acid was oxidized in part to 2-[4-carboxy-2-ethoxyphenoxy]-propane-1-ol), and (iii) reduction of carboxyl groups (older cultures) (e.g., 3-ethoxy-4-isopropoxybenzoic acid was reduced to 3-ethoxy-4-isopropoxybenzaldehyde and 3-ethoxy-4-isopropoxybenzyl alcohol). Some ethers (e.g., tri-O-methyl gallic acid and glycerol-beta-[4-carboxy-2-ethoxyphenyl]-ether) were not affected. The dealkylations and hydroxylations indicate that the fungus has a relatively nonspecific mechanism for oxygenating various 4-alkoxyl groups of alkoxybenzoic acids; no evidence for oxygenation of 3-alkoxyl groups was obtained. Hydroxylation products were generally degraded further, probably via dealkylation. The vanillic acid and 3-ethoxy-4-hydroxybenzoic acid formed by dealkylations were readily metabolized. Although the isopropyl ether of syringic acid was hydroxylated to 2-(4-carboxy-2, 6-dimethoxyphenoxy)-propane-1-ol, neither this compound nor the parent isopropyl ether was dealkylated; syringic acid itself was only slowly and incompletely metabolized. The relationship of these results to lignin degradation is discussed.

Basidiomycota↗

Degradation of methoxylated benzoic acids by a Nocardia from a lignin-rich environment: significance to lignin degradation and effect of chloro substituents.

Strain A81 of Nocardia corallina hydroxylates or demethylates p-anisic acid to p-hydroxybenzoic acid and isovanillic acid. It demethylates veratric acid to a mixture of vanillic and isovanillic acids. These are both demethylated to protocatechuic acid, which undergoes ring cleavage to afford beta-carboxy-cis-cis-muconic acid. The intermediacy of protocatechuic acid in the catabolic pathway of veratric acid was confirmed by blocking ring cleavage with an additional substituent in the ring: 5-chlorovanillic acid was demethylated to 5-chloro-protocatechuic acid, which accumulated. Chloro substituents in the ring of other methoxylated benzoic acids also arrested their normal metabolism by the Nocardia: an ortho-chloro substituent thwarted both demethylation and ring-opening. ortho-Hydroxylation of p-methoxybenzoic acid to isovanillic acid was unaffected by a chlorine ortho to the methoxyl group. Washed whole cells of veratric acid-grown N. corallina A81 produced no detected structural changes in an isolated lignin. The implications of this observation are discussed.

Benzoates↗