Microbial metabolism of quinoline and related compounds. II. Degradation of quinoline by Pseudomonas fluorescens 3, Pseudomonas putida 86 and Rhodococcus spec. B1

Quinoline catabolism was investigated with different bacterial strains, able to use quinoline as sole source of carbon, nitrogen and energy. Some degradation products of quinoline were isolated from the culture fluids and identified. With Pseudomonas fluorescens and Pseudomonas putida we found 2-oxo-1,2-dihydroquinoline, 8-hydroxy-2-oxo-1,2-dihydroquinoline, 8-hydroxycoumarin and 2,3-dihydroxyphenylpropionic acid as intermediates. With a Rhodococcus strain 2-oxo-1,2-dihydroquinoline, 6-hydroxy-2-oxo-1,2-dihydroquinoline, a red meta-cleavage product and a blue fluorescent compound were isolated. The red compound was identified as 5-hydroxy-6-(3-carboxy-3-oxopropenyl)-1H-2-pyridone. From this the blue fluorescent azacoumarin 2H-pyrano-2-one-[3,2b]-5H-6-pyridone is formed by chemical decomposition. Therefore it can be considered a by-product of quinoline-degradation in Rhodococcus spec. With the present results two different degradation pathways for quinoline in different microorganisms are proposed.     

Microbial metabolism of quinoline and related compounds. XI. Degradation of quinoline-4-carboxylic acid by Microbacterium sp. H2, Agrobacterium sp. 1B and Pimelobacter simplex 4B and 5B.

From soil enrichment cultures four strains, using quinoline-4-carboxylic acid as sole source of energy and carbon, have been isolated. According to their physiological properties these bacteria have been identified as Microbacterium sp. designated H2, as Agrobacterium sp. designated 1b and Pimelobacter simplex designated 4B and 5B. Metabolites of the degradation pathway of quinoline-4-carboxylic acid have been isolated and identified. With Pimelobacter simplex 4B and 5B 2-oxo-1,2-dihydroquinoline-4-carboxylic acid and 8-hydroxycoumarin-4-carboxylic acid were isolated. The Agrobacterium strain accumulated 2-oxo-1,2-dihydroquinoline-4-carboxylic acid and 2-oxo-1,2,3,4-tetrahydroquinoline-4-carboxylic acid in the media during growth; with Microbacterium sp. H2 we only found 8-hydroxycoumarin-4-carboxylic acid. With mutants of Microbacterium sp. H2 which were induced with N-methyl-N'-nitro-N-nitrosoguanidine we found 2-oxo-1,2-dihydroquinoline-4-carboxylic acid, 8-hydroxy-coumarin-4-carboxylic acid and 2,3-dihydroxyphenyl-succinic acid.     

苯酚高效降解菌的筛选和降解特性的研究

从天津市煤气厂的活性污泥中筛选、分离得到一株高效苯酚降解菌。经BIOLOG细菌自动鉴定系统及16SrDNA鉴定,该菌株为粪产碱杆菌(Alcaligenesfaecalis)。苯酚降解实验证实,该菌能在76h内完全降解1600mg·L-1的苯酚,并且随着苯酚浓度的增加,底物抑制作用增强,细胞得率下降。     

热凝胶的高产策略及功能研究进展

热凝胶（curdlan）又名β-（1→3）-D-葡聚糖,在食品、医药、保健品等领域具有重要的应用潜力。由粪产碱杆菌（Alcaligenes faecalis）或土壤杆菌（Agrobacterium sp．）生物合成的热凝胶以成本低廉、提取与纯化技术成熟而成为研究热点。笔者主要针对提高热凝胶产量的菌株改造和发酵控制策略以及热凝胶生物活性的研究进展和相关专利进行综述。     

Alcaligenes faecalis subsp. parafaecalis subsp. nov., a bacterium accumulating poly-beta-hydroxybutyrate from acetone-butanol bioprocess residues

The authors have previously isolated a solvent tolerant bacterium, strain G(T), (T = type strain) capable to convert acetone-butanol bioprocess residues into poly-beta-hydroxybutyrate. Strain G(T) was initially identified as Alcaligenes spp by standard bacteriological tests. In this study the taxonomic position of the bacterium was investigated in detail. The 165 rDNA sequence analysis, the G + C content of DNA (56 mol%) and the presence of ubiquinone Q-8 confirmed strain G(T) as a representative of the genus Alcaligenes. In the polyamine pattern of the bacterium putrescine and cadaverine were detected, but only trace amounts of 2-hydroxyputrescine. The extremely low content of 2-hydroxyputrescine is remarkable, since this unique diamine is a common marker for beta-proteobacteria. Phylogenetic analyses of 16S rDNA demonstrated that Alcaligenes sp. G(T) is most closely related to the species Alcaligenes faecalis (99.6% sequence similarity to A. faecalis HR4 and 98.7% sequence similarity to A. faecalis [ATCC 8750T = DSM 30030T]. On the basis of DNA-DNA relatedness (56% similarity), the unique polyamine pattern, the physiological and biochemical differences strain G(T) could be distinguished from the species A. faecalis. Therefore, a new subspecies for the species Alcaligenes faecalis is proposed; Alcaligenes faecalis subsp. parafaecalis subsp. nov.     

Spontaneous mutation of polysaccharide production in Alcaligenes faecalis var. myxogenes 10C3.

Alcaligenes faecalis var. myxogenes 10C3, which produces large amounts of succinoglucan and small amounts of curdlan, was genetically unstable and mutated spontaneously to a form producing more curdland than succinoglucan when stocked on nutrient agar slants. The mutation occurred in the absence of cell division when the cells were incubated in saline and was enhanced by treatment with N-methyl-N'-nitro-N-nitrosoguanidine, ethyl methane sulfonate, or ultraviolet light. Mutant strains were genetically stable and did not revert spontaneously for at least 1 year when stocked on nutrient agar slants.     

Selection and identification of bacteria isolated from waste crude oil with polycyclic aromatic hydrocarbons removal capacities

Fifteen bacterial strains isolated from solid waste oil samples were selected due to their capacity of growing in the presence of hydrocarbons. The isolates were identified by PCR of the 16S rDNA gene using fD1 and rD1 primers. The majority of the strains belonged to genera Bacillus, Bacillus pumilus (eight strains) and Bacillus subtilis (two strains). Besides, three strains were identified as Micrococcus luteus, one as Alcaligenes faecalis and one strain as Enterobacter sp. Growth of the above-mentioned strains in mineral liquid media amended with naphthalene, phenanthrene, fluoranthene or pyrene as sole carbon source was studied and our results showed that these strains can tolerate and remove different polycyclic aromatic hydrocarbons that may be toxic in the environment polluted with hydrocarbons. Finally, the capacity of certain strains to emulsify octane, xilene, toluene, mineral oil and crude oil, and its ability to remove hydrocarbons, look promising for its application in bioremediation technologies.     

生姜根际茎腐病拮抗菌的筛选及鉴定

从生姜根际土样分离获得的86个细菌分离物中,通过离体拮抗试验,筛选出2株对生姜茎腐病有良好拮抗效果的细菌,编号分别为JF24和JF3。通过对它们进行形态学观察、生理生化测定以及16S rRNA序列分析,初步确定,JF24为荧光假单胞菌(Pseudomonas fluorescens),JF3为粪产碱菌(Alcaligenes faecalis)。JF24和JF3的16S rRNA序列的GenBank登录号分别为FJ999730和FJ999731。     

The Properties and Classification of the Predominant Bacteria in Rotten Eggs

A collection of 226 strains of bacteria was assembled from eggs which had rotted on the premises of the producer and from others which had been allowed to rot in the laboratory at 10, 20 or 30°. A majority of the eggs had a mixed infection. All but 8 of the isolates were Gram negative rods. The predominant types were Alcaligenes faecalis, Aeromonas liquefaciens, Proteus vulgaris, Cloaca spp., Citrobacter sp. and Pseudomonas fluorescens. A nonpigmented pseudomonad could not be identified with any of the species included in Pseudomonas.     

Microbial metabolism of quinoline and related compounds. IX. Degradation of 6-hydroxyquinoline and quinoline by Pseudomonas diminuta 31/1 Fa1 and Bacillus circulans 31/2 A1

Two strains, using 6-hydroxyquinoline as sole source of energy, carbon and nitrogen, have been isolated. These bacteria, designated 31/1 Fa1 and 31/2 A1, are also able to degrade quinoline. According to their physiological properties strain 31/1 Fa1 has been identified as Pseudomonas diminuta and strain 31/2 A1 as Bacillus circulans. 6-Hydroxy-2-oxo-1,2-dihydroquinoline was found as intermediate in the degradation of 6-hydroxyquinoline and quinoline. 2-Oxo-1,2-dihydroquinoline was the first metabolite in the degradation of quinoline.     

Identification of dimethyl disulfide-forming bacteria isolated from activated sludge

Twenty-four strains with high dimethyl disulfide (DMDS)-forming ability were isolated from activated sludge and identified to the genus level. These bacteria were classified into four groups (A, B, C, and D) by the API ZYM System (API System S.A., Montalieu, France). Group A (three strains) was identified as genus Lactobacillus by the API 20B System, by the method of Cowan and Steel, and by production of lactic acid as confirmed by gas-liquid chromatography. Group B (eight strains) was identified as genus Corynebacterium by API 20B and the Cowan and Steel method. Group C (one strain) was suggested to belong to genus Corynebacterium by the API 20B System. Group D (12 strains) was identified as genus Pseudomonas or Alcaligenes by the API 20B System, as genus Alcaligenes by the Cowan and Steel method, and as Achromobacter group Vd by the API 20NE System. However, on the basis of guanine-plus-cytosine contents in DNA and form of flagella, these strains were identified as genus Pseudomonas. Formation of DMDS from DL-methionine and S-methyl-L-cysteine was tested. DMDS-forming bacteria isolated from activated sludge formed DMDS from both precursors. In genus Pseudomonas, P. aeruginosa could not form DMDS from either precursor, but P. acidovorans, P. alcaligenes, P. pseudoalcaligenes, and P. testosteroni formed DMDS. In genus Alcaligenes, A. denitrificans subsp. xylosoxydans, A. denitrificans subsp. denitrificans, A. faecalis, and A. odorans formed DMDS from both precursors. Achromobacter group Vd formed DMDS from S-methyl-L-cysteine, but could not from DL-methionine.     

Spontaneous mutation of polysaccharide production in Alcaligenes faecalis var. myxogenes 10C3

Alcaligenes faecalis var. myxogenes 10C3, which produces large amounts of succinoglucan and small amounts of curdlan, was genetically unstable and mutated spontaneously to a form producing more curdland than succinoglucan when stocked on nutrient agar slants. The mutation occurred in the absence of cell division when the cells were incubated in saline and was enhanced by treatment with N-methyl-N'-nitro-N-nitrosoguanidine, ethyl methane sulfonate, or ultraviolet light. Mutant strains were genetically stable and did not revert spontaneously for at least 1 year when stocked on nutrient agar slants.     

Microbial oxidation of dimethylnaphthalene isomers.

Three bacterial strains, identified as Alcaligenes sp. strain D-59 and Pseudomonas sp. strains D-87 and D-186, capable of growing on 2,6-dimethylnaphthalene (2,6-DMN) as the sole source of carbon and energy were isolated from soil samples. 2,6-Naphthalene dicarboxylic acid was formed in the culture broths of these three strains grown on 2,6-DMN. In addition, 2-hydroxymethyl-6-methylnaphthalene and 6-methylnaphthalene-2-carboxylic acid were detected in the culture broth of strain D-87. Strain D-87 grew well on 1,2-, 1,3-, 1,4-, 1,5-, 2,3-, and 2,7-DMN as the sole source of carbon and energy and accumulated 2-methylnaphthalene-3-carboxylic acid and 2,3-naphthalene dicarboxylic acid from 2,3-DMN, 4-methylnaphthalene-1-carboxylic acid from 1,4-DMN, and 7-methylnaphthalene-2-carboxylic acid from 2,7-DMN.     

Microbial Metabolism of Quinoline by Comamonas sp.

An aerobic bacterial strain which can use quinoline as the sole carbon and energy source has been isolated from activated sludge and identified as Comamonas sp. The microbial metabolism of quinoline by this strain has been investigated. A pH 8 and a temperature of 30 °C were the optimum degradation conditions of quinoline. Five intermediates including 2-oxo-1,2-dihydroquinoline, 5-hydroxy-6-(2-carboxyethenyl)-1H-2-pyridone, 6-hydroxy-2-oxo-1,2-dihydroquinoline, 5,6-dihydroxy-2-oxo-1,2-dihydroquinoline, and 8-hydroxy-2-oxo-1,2-dihydroquinoline were found during quinoline biodegradation. The presence of these intermediates suggested that at least two pathways were involved for quinoline degradation by Comamonas sp. and a reasonable degradation route was proposed to account for the intermediates observed. This revised version was published online in July 2006 with corrections to the Cover Date.     

Molecular analysis of maleate cis-trans isomerase from thermophilic bacteria

Several strains of thermophilic bacteria containing maleate cis-trans isomerase were isolated from soil samples and identified as Bacillus stearothermophilus, Bacillus circulans, Bacillus brevis, and Deleya halophila. The maleate cis-trans isomerase was purified and characterized from one of the isolated strains, B. stearothermophilus MI-102. The purified enzyme of strain MI-102 showed higher thermal stability than the enzyme of a mesophile, Alcaligenes faecalis IFO13111. The seven maleate cis-trans isomerase genes (maiA) of thermophile were cloned and sequenced. B. stearothemophilus MI-102 MaiA has 67% amino acid identity with A. faecalis MaiA. All eight amino acid sequences of maiA gene products had significant conserved regions containing cysteine residues, which were previously suggested to be involved in an active site of the enzyme. To probe the catalytic mechanism, three cysteine residues in the conserved regions of A. faecalis MaiA were replaced with serine by site-directed mutagenesis. The results suggest that Cys80 and Cys198 play important roles in the enzyme activity.     

生物絮凝剂产生菌的分离与鉴定

通过多点采样,多次反复筛选,从旱田土壤中筛选出1株具有高絮凝活性的细菌。细菌生长过程均可产生具有絮凝作用的胞外分泌物,经实验对内蒙天然碱碱泥具有高絮凝作用。通过对菌株的个体形态特征、菌落形态特征、运动性进行观察;同时对其接触酶、氧化酶等生理生化指标进行了测试,最终确定为粪产碱杆菌（Alcaligenes faecalis)。     

A note on the isoprenoid quinone content of Bordetella avium and related species

The isoprenoid quinone content of isolates of Bordetella avium (four strains), Alcaligenes faecalis (one strain), Bordetella bronchiseptica (one strain) and a Bordetella avium -like organism (four strains) was determined by reverse-phase high-performance liquid chromatography. All the isolates contained ubiquinones with eight isoprene units as the major component. No menaquinones were detected.     

Antifungal effect of a heterotrophic nitrifier Alcaligenes faecalis

Alcaligenes faecalis suppressed the growth of 11 strains of fungal plant pathogens in vitro. When it was cultivated on a synthetic medium containing (NH4)2 SO4 as the sole nitrogen source, NH2OH, NO2- and NO3- were produced, indicating that heterotrophic nitrification was occurring. The suppressive effect of A. faecalis on plant pathogens was due to its NH2OH produced. © Rapid Science Ltd. 1998     

A note on the isoprenoid quinone content of Bordetella avium and related species

The isoprenoid quinone content of isolates of Bordetella avium (four strains), Alcaligenes faecalis (one strain), Bordetella bronchiseptica (one strain) and a Bordetella avium-like organism (four strains) was determined by reverse-phase high-performance liquid chromatography. All the isolates contained ubiquinones with eight isoprene units as the major component. No menaquinones were detected.     

Transfer and expression of Klebsiella nif genes in Alcaligenes faecalis, a nitrogen-fixing bacterium associated with rice root

Plasmid pRD1 carrying Klebsiella nif genes was found to be transferable by conjugation from Escherichia coli JC5466 (pRD1) to Alcaligenes faecalis A-15 at a frequency of 5 X 10(-4). Nitrogenase activity of four A-15 (pRD1) strains tested was found to be higher than that of their parent A-15, as determined by the acetylene reduction assay. A-15-1 was a Nif- mutant derived from A-15. After mating with JC5466 (pRD1), the nitrogenase activity was restored. PRD1 was stable in A. faecalis and could be transferred to E. coli JC5466-1 by conjugation. The fact that Klebsiella nif genes carried in pRD1 can be expressed in A. faecalis makes it possible to use pRD1 as a tool for genetic analysis and genetic engineering of the nitrogen fixation system in A. faecalis.