Degradation of long chain n-alkanes by Mucorales

Summary The degradation pathways on n-dodecane and n-tridecane were studied in seven representative strains of five families of the order Mucorales. Using thin-layer chromatographic, gas-liquid chromatographic and mass spectrometric methods, extracellular oxidation products of the relevant n-alkanes could be isolated and identified. All strains tested exhibited a formation of isomeric primary and secondary alcohols, isomeric ketones and monoic acids with chain length equivalent to the n-alkanes in the substrates, proving that in the order Mucorales both a terminal and a subterminal oxidation pathway are realized.     

Degradation of long chain n-alkanes by mucorales

Summary Extracellular oxidation products isolated from the substrates ofMortierella isabellina (CBS 224. 35) grown on n-dodecane and n-tridecane were primary and secondary isomeric alcohols, isomeric ketones, aldehydes and isomeric esters with the same numbers of carbon atoms presented in the used n-alkanes as detected by combined glass capillary gas chromatography — mass spectrometry. All esters were identified which theoretically could originate in the isomeric ketones by a reaction mechanism resembling a Baeyer-Villinger-type oxidation.     

Degradation of long chain n-alkanes by disrupted cells ofChlorella vulgaris

Summary The alkane oxidation byChlorella vulgaris is improved by disruption of the cells. Although living cells are not able to attack n-dodecane, disrupted cells produced detectable amounts of oxidation products. The amount of isomeric alcohols and ketones of n-tridecane was nearly double the sum found in living cells, whereas the equilibrium was shifted to the ketones. With n-tetradecane and n-pentadecane only the amount of ketones increased.     

Degradation of long chain alkanes by bacilli

Summary Among 14 different types of bacillus and 100 isolates from soil samples, no bacillus was able to assimilate C₁₄-C₁₈ alkanes as sole carbon source. In contrast to these results, five types of bacillus were substantially able to oxidize alkanes in the presence of other carbon sources.All five strains tested formed relatively high amounts of secondary alcohol in contrast to low amounts of ketones and traces of primary alcohols with chain length equivalent to the n-alkane in the substrate, showing that the degradation pathway in bacilli is mainly subterminal.This conclusion is also supported by comparison of the extracellular fatty acids of cultures of bacilli grown on n-tridecane and on n-tetradecane supplemented with other carbon sources and on the same carbon sources without alkane.     

Degradation of long chain alkanes by bacilli

Summary Five different species of bacilli tested at 30°C degrade hydrocarbons by cooxidation. Subterminal degradation pathway of these bacilli was proved by estimating isomeric esters, developed from Baeyer-Villiger oxidation of ketones, and also by finding diols, ketols, and diketones. Comparing the composition of fatty acids in lipids of bacilli grown with and without alkane as well as growing them on labeled n-tridecane suggested a subterminal, partly a di-subterminal pathway.It was also shown that Bacillus stearothermophilus cannot grow on alkane as sole carbon source even in a thermophilic temperature range and that oxidation products are formed in amounts twenty times less when grown on cooxidation medium at 50°C, compared to product formation on the same medium at 30°C.     

Degradation of long chain alkanes by bacilli

Summary Degradation of tetradecane by two thermophilic Bacillus strains was investigated. It was found that these strains do not grow on this hydrocarbon as sole carbon source but degrade tetradecane in cooxidation cultures partly via monoterminal pathway.     

Degradation of long-chain n-alkanes by mucorales

Summary The composition of fatty acids in the lipids ofAbsidia spinosa, Cunninghamella echinulata, andMortierella isabellina was determined after growth on glucose and some n-alkanes. Mycelia metabolizing odd-carbon n-alkanes contained, among others, mono- and polyenoic heptadecanoic acids, that is, C_17:1(9)-, C_{17:2(9, 12)}-, and C_{17:3(6, 9, 12)}-acids. From the amounts of the isolated lipids it can be deduced thatAbsidia andCunninghamella may mainly degrade the alkanes in a terminal oxidation pathway whileMortierella prefers a subterminal way.Dedicated to Professor Dr. L. Acker on the occasion of his 65th birthday     

Degradation of long-chain n-alkanes by the yeast Candida maltosa

Summary Microsomal membrane fractions of the yeast Candida maltosa were investigated with respect to their ability to catalyse the oxidation of n-alkanes, fatty alcohols and fatty acids. Analysis of intermediates of n-hexadecane oxidation led to the conclusion that monoterminal attack was predominant, whereas diterminal oxidation proceeded as a minor reaction. The oxidation of long-chain primary alcohols to the corresponding aldehydes occurred without addition of nicotinamide adenine dinucleotide (phosphate) [NAD(P)⁺] and was accompanied by stoichiometric oxygen consumption and hydrogen peroxide production, suggesting that an alcohol oxidase instead of an NAD(P)⁺-requiring alcohol dehydrogenase catalysed these reactions. As shown for n-hexadecane, the hydroxylation of palmitic acid was found to be carbon monoxide-dependent, indicating involvement of a cytochrome P-450 system, as in the case of n-alkane hydroxylation.     

Degradation of long-chain n-alkanes by the yeast Lodderomyces elongisporus

Summary Cells of the yeast Lodderomyces elongisporus, precultured on glycerol, were incubated with long-chain n-alkanes. The results whow that monoterminal alkane oxidation is the main pathway of alkane degradation in the investigated yeast. The amount of diterminal activity is negligible, while subterminal degradation did not occur at all.Fatty acids were the first detectable intermediates. Using different n-alkanes, in every case the fatty acids with substrate chain length predominated in the cells. The formation of radioactive fatty acids from (1-¹⁴C)-hexadecane was time-dependent and indicated that desaturation elongation and -oxidation occurred.Extracellularly, the fatty acid pattern was similar, except for the additional presence of fatty acid methyl esters and the prevalence of octadecenoic acid after growth of cells on n-hexadecane.     

Fast biodegradation of long chain n-alkanes and crude oil at high concentrations with Rhodococcus sp. Moj-3449

Biodegradation of long chain n-alkanes and crude oil with fast rate and high concentration are desirable for bioremediation, especially in heavily oil-polluted areas, and enhanced oil recovery. We discovered Rhodococcus sp. Moj-3449 with such unique abilities by screening microorganisms for the growth on n-hexadecane at 30 mg/mL. The new strain grew very fast on 120 mg/mL of n-hexadecane giving a cell density of 14.7 g cdw/L after only 2 days’ incubation. During the growth with this strain, the oil–water phases were rapidly emulsified, giving rise to tolerance to high alkane concentration (250 mg/mL) and fast growth rate of 0.10–0.20 h^?1 for alkane concentration of 1–180 mg/mL. The degraded concentration of n-hexadecane increased linearly with the initial alkane concentration (1–250 mg/mL). Incubation on n-hexadecane at 250 mg/mL for 7 days gave a cell density of 13.5 g cdw/L and degraded 124 mg/mL of n-hexadecane. The strain grew also fast on n-dodecane (C12), n-tetradecane (C14), and n-octadecane (C18), with degradation preference of C14 (=C16) > C12 > C18. Different from many alkane-degrading strains, Rhodococcus sp. Moj-3449 was found to have subterminal oxidation pathway. Rhodococcus sp. Moj-3449 degraded also crude oil fast at 60–250 mg/mL, with a wide range of n-alkanes (C10–C35) as substrates in which C14–C19 are preferred. The degradation ability increased with initial oil concentration from 60 to 150 mg/mL and slightly decreased afterwards. Incubation on 150 mg/mL of crude oil for 7 days degraded 37% of n-alkanes. The outstanding ability of rapidly degrading long chain n-alkanes and crude oil at high concentration makes Rhodococcus sp. Moj-3449 potentially useful for bioremediation and microbial enhanced oil recovery.     

Degradation of n-alkanes by Candida parapsilosis and Penicillium frequentans immobilized on granular clay and aquifer sand

Summary The immobilization intensity of cells of Penicillium frequentans and Candida parapsilosis on materials such as granular clay, granular clay + aquifer sand and aquifer sand alone, was followed by scanning electron microscopy (SEM). The results demonstrate that the granular clay was the best adsorbent for both organisms, followed by the mixture of both granular clay and aquifer sand. Poor adhesion of cells was detected on using aquifer sand alone with C. parapsilosis.The highest degree of degradation of the alkane mixture (C₁₂–C₁₈) used was achieved by cells immobilized on granular clay, followed by those cells adsorbed on clay and sand. The weakest degradation was observed with cells immobilized on the sand alone.     

Degradation of long-chain n-alkanes (C36 and C40) by Pseudomonas aeruginosa strain WatG

Pseudomonas aeruginosa strain WatG was unable to utilize either n-hexatriacontane (C₃₆) or n-tetracontane (C₄₀), which are both insoluble in a mineral salts medium (MSM), as a sole carbon source. However, when C₃₆ and C₄₀ were added to MSM containing crude oil, more than 25% of each of the compounds was degraded by this strain after 2 weeks at 30 °C. These results demonstrate that P. aeruginosa strain WatG has the ability to degrade long-chain alkanes up to C₄₀, when they are solubilized by crude oil components.     

Platelet aggregation is inhibited by long chain acyl-CoA

Oleoyl coenzyme A and other acyl-CoA derivatives inhibited ADP or thrombin-induced aggregation of platelets. Arachidonic acid-induced aggregation was also inhibited, but not the slower aggregation caused by 1-oleoyl-2-acetylglycerol or tetradecanoyl-phorbol-13-acetate. Coenzyme A and free fatty acids had little or no effect, and transfer of labeled oleate from oleoyl Co-A to other lipid classes was not detected. Because acyl Co-A compounds have recently been shown to modulate protein kinase C activity, acyl Co-A may provide a useful tool for investigating activation sequences in platelets and other membranes.     

Degradation of very long chain dicarboxylic polyunsaturated fatty acids in mouse hepatocytes, a peroxisomal process

Polyunsaturated fatty acids can be omega-oxidized to dicarboxylic polyunsaturated fatty acids (DC-PUFA), bioactive compounds which cause vasodilatation and activation of PPARalpha and gamma. DC-PUFA can be shortened by beta-oxidation, and to determine whether mitochondria and/or peroxisomes are responsible for this degradation 20-carboxy-[1-(14)C]-eicosatetraenoic acid (20-COOH-AA) was synthesized and given to hepatocytes from mouse models with peroxisomal dysfunctions. In contrast to wild type cells, hepatocytes from mice with liver-selective elimination of peroxisomes, due to Pex5p deficiency, failed to produce (14)CO(2) and labeled acid-soluble oxidation products, indicating that peroxisomes are involved in the degradation of 20-COOH-AA. Subsequently, the oxidation of 20-COOH-AA was analyzed in hepatocytes lacking multifunctional protein 1 (MFP1) or MFP2, key enzymes of the peroxisomal beta-oxidation. Degradation of 20-COOH-AA was partially impaired in MFP1, but not in MFP2 knockout hepatocytes. Taken together, peroxisomes and not mitochondria are the site of beta-oxidation of DC-PUFA, and MFP1 is involved in this process.     

Inhibition of chymase activity by long chain fatty acids

The activity of chymase was markedly inhibited by fatty acids with carbon chain lengths of 14-22 at doses greater than 0.02 microM, irrespective of the number of double bonds. Cis acids with a carbon chain length of 18, such as stearic acid, oleic acid, linoleic acid, and linolenic acid were potent inhibitors, whereas the trans isomer of oleic acid, elaidic acid, showed less inhibitory activity. The extent of inhibition by oleyl alcohol was almost the same as that by oleic acid, suggesting that the acid moiety itself was not necessary for the inhibition; but a fatty acid with a terminal functional amide, oleamide, showed little inhibitory activity. The inhibition was noncompetitive and was reversible, and the Ki value of oleic acid was 2.7 microM. Stearic acid and oleic acid inhibited all chymotrypsin-type serine endopeptidases tested. The ID50 values of these fatty acids for atypical mast cell protease were higher than those for the other chymotrypsin-type serine endopeptidases tested. Other proteases, such as papain, trypsin, collagenase, and carboxypeptidase A, except cathespin D, were not affected by stearic or oleic acid.     

Degradation of n-alkanes and polycyclic aromatic hydrocarbons in petroleum by a newly isolated Pseudomonas aeruginosa DQ8

A bacterial isolate, designated as DQ8, was found capable of degrading diesel, crude oil, n-alkanes and polycyclic aromatic hydrocarbons (PAHs) in petroleum. Strain DQ8 was assigned to the genus Pseudomonas aeruginosa based on biochemical and genetic data. The metabolites identified from n-docosane as substrate suggested that P. aeruginosa DQ8 could oxidize n-alkanes via a terminal oxidation pathway. P. aeruginosa DQ8 could also degrade PAHs of three or four aromatic rings. The metabolites identified from fluorene as substrate suggested that P. aeruginosa DQ8 may degrade fluorene via two pathways. One is monooxygenation at C-9 of fluorene, and the other is initiated by dioxygenation at C-3 and C-4 of fluorene. P. aeruginosa DQ8 should be of great practical significance both in bioremediation of oil-contaminated soils and biotreatment of oil wastewater.