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New developments in oxidative fermentation

APPLIED MICROBIOLOGY AND BIOTECHNOLOGY 2003年第6期60卷 643-653页

作者： Adachi, O Moonmangmee, D Toyama, H Yamada, M Shinagawa, E Matsushita, K Yamaguchi Univ Fac Agr Dept Biol Chem Yamaguchi 7538515 Japan King Mongkuts Univ Technol Thonburi Fac Sci Dept Microbiol Bangkok 10140 Thailand Ube Natl Coll Technol Fac Biotechnol Dept Biol & Chem Engn Ube Yamaguchi 7558555 Japan

Oxidative fermentations have been well established for a long time, especially in vinegar and in L-sorbose production. Recently, information on the enzyme systems involved in these oxidative fermentations has accumulated and new developments are possible based on these findings. We have recently isolated several thermo-tolerant acetic acid bacteria, which also seem to be useful for new developments in oxidative fermentation. Two different types of membrane-bound enzymes, quinoproteins and flavoproteins, are involved in oxidative fermentation, and sometimes work with the same substrate but produce different oxidation products. Recently, there have been new developments in two different oxidative fermentations, D-gluconate and D-sorbitol oxidations. Flavoproteins, D-gluconate dehydrogenase, and D-sorbitol dehydrogenase were isolated almost 2 decades ago, while the enzyme involved in the same oxidation reaction for D-gluconate and D-sorbitol has been recently isolated and shown to be a quinoprotein. Thus, these flavoproteins and a quinoprotein have been re-assessed for the oxidation reaction. Flavoprotein D-gluconate dehydrogenase and D-sorbitol dehydrogenase were shown to produce 2-keto-D-gluconate and D-fructose, respectively, whereas the quinoprotein was shown to produce 5-keto-D-gluconate and L-sorbose from D-gluconate and D-sorbitol, respectively. In addition to the quinoproteins described above, a new quinoprotein for quinate oxidation has been recently isolated from Gluconobacter strains. The quinate dehydrogenase is also a membrane-bound quinoprotein that produces 3-dehydroquinate. This enzyme can be useful for the production of shikimate, which is a convenient salvage synthesis system for many antibiotics, herbicides, and aromatic amino acids synthesis. In order to reduce energy costs of oxidative fermentation in industry, several thermotolerant acetic acid bacteria that can grow up to 40degreesC have been isolated. Of such isolated strains, some thermotoleran

关键词：醋酸/代谢醋杆菌/代谢不动杆菌属/代谢细菌蛋白质类/生理学发酵黄素腺嘌呤二核苷酸/代谢黄素蛋白类/生理学果糖/代谢葡糖酸盐类/代谢葡萄糖菌/代谢热工业微生物学酮类/代谢 L-艾杜糖醇-2-脱氢酶/代谢氧化还原 PQQ辅因子奎尼酸/类似物和衍生物奎尼酸/代谢喹诺酮类/代谢醌类/代谢莽草酸/代谢山梨醇/代谢丁糖类/代谢

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Effect of dispersing oil phase on the biodegradability of a solid alkane dissolved in non-biodegradable oil

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APPLIED MICROBIOLOGY AND BIOTECHNOLOGY 2002年第4-5期59卷 574-579页

作者： Hori, K Matsuzaki, Y Tanji, Y Unno, H Tokyo Inst Technol Grad Sch Biosci & Biotechnol Midori Ku Yokohama Kanagawa 2268501 Japan

Acinetobacter sp. CR was grown on a model oil, which consisted of an inert oil matrix of pristane with n-heneicosane dissolved in it as the sole carbon source, in a stirred-tank bioreactor. This bacterium takes up substrates from the oil phase by direct contact with the oil phase. A previously established mathematical model was applied to reveal the effect of agitation conditions on the growth and n-alkane degradation kinetics of the bacterium. Higher impeller speed resulted in both lower microbial growth and lower n-alkane degradation rate of the bacterium, although it increased the specific surface area of the oil, which was measured by a previously developed device. This result was due to the decreased number of cells adhering to the oil surface, i.e., intense agitation inhibited the adhesion of cells to the oil surface. The addition of a surfactant below a critical micelle concentration (CMC) inhibited the degradation of n-heneicosane dissolved in pristane, although the biodegradability of the substrate recovered gradually with the increase in the dose of surfactant over CMC. The results suggest that efforts to increase the specific surface area of the oil phase have the undesirable result of inhibiting oil degradation when the dominant microbial degraders take up substrates in oil by direct contact with the oil.

关键词：不动杆菌属/药物作用不动杆菌属/生长和发育不动杆菌属/代谢烷烃类/化学烷烃类/代谢细菌黏附生物降解环境/药物作用生物反应器培养基乳状剂动力学模型生物学辛苯昔醇/药理学溶解度表面特性表面活性剂/药理学萜类/化学

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Isolation of an Acinetobacter sp ST-550 which produces a high level of indigo in a water-organic solvent two-phase system containing high levels of indole

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APPLIED MICROBIOLOGY AND BIOTECHNOLOGY 2002年第4期58卷 543-546页

作者： Doukyu, N Nakano, T Okuyama, Y Aono, R Tokyo Inst Technol Grad Sch Biosci & Biotechnol Dept Biol Informat Midori Ku Yokohama Kanagawa 2268501 Japan

Acinetobacter sp. strain ST-550 was isolated from fumus soil as an efficient indigo producer in the presence of organic solvent. The minimum inhibitory concentration of indole was 0.4 mg/ml for ST-550. ST-550 produced only a small amount of indigo (less than 0.01 mug/ml) when grown in the presence of indole at concentrations of 0.05 to 0.3 mg/ml without any organic solvent. However, ST-550 produced indigo effectively when grown in the presence of a large: volume of diphenylmethane and a high level of indole;optimized conditions were 3 ml of a medium containing 0.3 ml diphenylmethane and 2.7 mg indole. Under these conditions, ST-550 produced 0.88 mg indigo (292 mug/ml medium).

关键词：不动杆菌属/分类不动杆菌属/分离和提纯不动杆菌属/代谢生物降解环境培养基发酵吲哚类/化学吲哚类/代谢土壤微生物学溶剂

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Solubilization of polyaromatic hydrocarbons by recombinant bioemulsifier AlnA

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APPLIED MICROBIOLOGY AND BIOTECHNOLOGY 2002年第4-5期59卷 580-584页

作者： Toren, A Ron, EZ Bekerman, R Rosenberg, E Tel Aviv Univ Dept Mol Microbiol & Biotechnol IL-69978 Ramat Aviv Israel

AlnA is the protein responsible for the emulsifying and solubilizing activity of the Acinetobacter radioresistens KA53 bioemulsifier alasan. AlnA was produced in Escherichia coli, purified to homogeneity and then used to measure the enhanced solubility of 12 polyaromatic hydrocarbons (PAHs). The amount of PAH solubilized was directly proportional to AlnA concentration. The ratio of PAH solubilized by 40 mug/ml AlnA compared to that soluble in the aqueous buffer varied greatly, from 4 (fluorene) to 81 (hexylbenzylcyclosilane). Calculations of moles PAH solubilized per mole AlnA yielded values from 4.3 (hexylphenylbenzene) to 55.8 (1,10-phenanthrolene). There was no obvious relationship between the amount of PAH solubilized and its molecular weight or intrinsic solubility. Native gel electrophoresis indicated that AlnA formed hexamers in the presence of PAHs. With molar ratios of fluorene to AlnA of 0.75 or less, only the monomer was observed, whereas at ratios of 7.5 or higher, only the hexamer was detected. At an intermediate molar ratio of 2.6, both monomer and hexamer appeared. The data indicate that PAHs are initially solubilized by binding to the monomeric form of AlnA, and as the amount bound increases above one molecule PAH per AlnA, the protein aggregates to form a specific oligomer of 5-8 monomers which allows for the binding and solubilization of more PAH.

关键词：不动杆菌属/遗传学不动杆菌属/代谢细菌外膜蛋白质类/遗传学细菌外膜蛋白质类/代谢细菌蛋白质类/代谢辅药芴类/代谢烃类芳香/化学烃类芳香/代谢多糖类细菌/代谢重组蛋白质类/遗传学重组蛋白质类/代谢溶解度

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4-hydroxybenzoate uptake in an isolated soil Acinetobacter sp.

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CURRENT MICROBIOLOGY 2000年第1期40卷 34-39页

作者： Allende, JL Gibello, A Fortún, A Mengs, G Ferrer, E Martín, M Univ Complutense Madrid Fac Vet Dept Bioquim & Biol Mol 4 E-28040 Madrid Spain Univ Complutense Madrid Fac Vet Dept Fis Aplicada E-28040 Madrid Spain Univ Complutense Madrid Fac Vet Dept Patol Anim & Microbiol E-28040 Madrid Spain

The isolated soil bacteria Acinetobacter strain BEM2 is able to utilize some xenobiotic aromatic compounds as a carbon source. In this study the metabolism of 4-hydroxybenzoate (4-HBA) by strain BEM2 was characterized. Degradation involved a meta-cleavage pathway yielding 3,4-dihydroxybenzoate (3,4-DHBA) as an intermediate and CO2 as the principal product from the C atoms in the aromatic ring. 4-HBA uptake was studied, and the kinetic parameters were determined. The uptake was shown to be directly coupled to ATP hydrolysis and its synthesis, according to the Mitchell chemiosmotic hypothesis.

关键词：不动杆菌属/酶学不动杆菌属/生长和发育不动杆菌属/代谢腺苷三磷酸/代谢需氧生活厌氧生活生物降解环境生物转运主动酶抑制剂/药理学动力学对羟苯甲酸类/代谢土壤微生物学

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Toluene vapour removal in a laboratory-scale biofilter

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APPLIED MICROBIOLOGY AND BIOTECHNOLOGY 2000年第2期54卷 248-254页

作者： Zilli, M Del Borghi, A Converti, A Univ Genoa Dept Chem & Proc Engn GB Bonino I-16145 Genoa Italy

A bench-scale biofilter with a 0.5-m high filter bed, inoculated with a toluene-degrading strain of Acinetobacter sp. NCIMB 9689, was used to study toluene removal from a synthetic waste air stream. Different sets of continuous tests were conducted at influent toluene concentrations ranging over 0.1-4.0 g m(-3) and at superficial gas velocities ranging over 17.8-255 m h(-1). The maximum volumetric toluene removal rate for the biofilter (242 g m(-3) h(-1)) was obtained at a superficial gas velocity of 127.5 m h(-1) (corresponding to a residence time of 28 s) and a toluene inlet concentration of 4.0 g m(-3). Under these operating conditions, toluene removal efficiency was only 0.238, which suggested that effective operation required higher residence times. Removal efficiencies higher than 0.9 were achieved at organic loads less than 113.7 g m(-3) h(-1). A macro-kinetic study, performed using concentration profiles along the bioreactor, revealed this process was limited by diffusion at organic loads less than 100 g m(-3) h(-1) and by biological reaction beyond this threshold.

关键词：不动杆菌属/代谢生物降解环境生物反应器扩散过滤气体动力学气味甲苯/代谢

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Phenotypic characterization and antibiotic resistance of Acinetobacter spp. isolated from aquatic sources

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JOURNAL OF APPLIED MICROBIOLOGY 1999年第5期87卷 659-667页

作者： Guardabassi, L Dalsgaard, A Olsen, JE Royal Vet & Agr Univ Dept Vet Microbiol DK-1870 Frederiksberg C Denmark

A total of 99 Acinetobacter isolates from sewage, freshwater aquaculture habitats, trout intestinal contents and frozen shrimps was characterized phenotypically and antibiotic susceptibility patterns determined. One group of genomic species, including Ac. johnsonii, Ac. lwoffi and spp. 15TU, was detected in all sample types and represented the majority of the isolates (n = 54). Isolates belonging to the Acb complex (Ac. calcoaceticus, Ac. baumannii and genomic species 3) were detected in sewage (n = 6) and frozen shrimps (n = 1), Ac. haemolyticus in frozen shrimps (n = 6) and trout intestinal contents (n = 2) and genomic species 11 in freshwater aquaculture habitats (n = 6) and trout intestinal contents (n = 1). Acinetobacter junii (n = 5), genomic species 10 (n = 2), 14BJ (n = 8) and 16BJ (n = 4) were only isolated from sewage. Acinetobacter isolates from sewage were generally more biochemically reactive and resistant to antimicrobial agents compared with isolates from other sample types. Different strains, often belonging to different genomic species, were isolated from sites situated upstream and downstream of the discharge point of a pharmaceutical plant. This finding supported the hypothesis that the waste effluent from the pharmaceutical plant was likely to cause a change in the distribution of Acinetobacter spp. by selecting and/or introducing antibiotic-resistant strains into the recipient sewers.

关键词：不动杆菌属/分类不动杆菌属/分离和提纯不动杆菌属/代谢阿莫西林/药理学细菌分型技术氯霉素/药理学环丙沙星/药理学 DNA 细菌/分析抗药性微生物淡水/微生物学庆大霉素类/药理学微生物敏感性试验土霉素/药理学表型污水/微生物学磺胺甲基异恶唑/药理学水微生物学

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Phenotypic expression of PCR-Generated random mutations in a Pseudomonas putida gene after its introduction into an Acinetobacter chromosome by natural transformation

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APPLIED AND ENVIRONMENTAL MICROBIOLOGY 1999年第4期65卷 1675-1680页

作者： Kok, RG Young, DM Ornston, LN Yale Univ Dept Biol New Haven CT 06520 USA

Localized sets of random point mutations generated by PCR amplification can be transferred efficiently to the chromosome of Acinetobacter ADP1 (also known as strain BD413) by natural transformation. The technique does not require cloning of PCR fragments in plasmids: PCR-amplified DNA fragments are internalized by cells and directly incorporated into their genomes by homologous recombination. Previously such procedures for random mutagenesis could be applied only to Acinetobacter genes affording the selection of mutant phenotypes, Here we describe the construction of a vector and recipient that allow for mutagenesis, recovery, and expression of heterologous genes that may lack a positive selection. The plasmid carries an Acinetobacter chromosomal segment interrupted by a multiple cloning site next to a kanamycin resistance marker. The insertion of heterologous DNA into the multiple cloning site prepares the insert as a target for PCR mutagenesis, PCR amplifies the kanamycin resistance marker and a flanking region of Acinetobacter DNA along with the insert of heterologous DNA. Nucleotide sequence identity between the flanking regions and corresponding chromosomal segments in an engineered Acinetobacter recipient allows homologous recombination of the PCR-amplified DNA fragments into a specific chromosomal docking site from which they can be expressed. The recipient strain contains only a portion of the kanamycin resistance gene, so donor DNA containing both this gene and the mutagenized insert can be selected by demanding growth of recombinants in the presence of kanamycin. The effectiveness of the technique was demonstrated with the relatively GC-rich Pseudomonas putida xylE gene. After only one round of PCR amplification (35 cycles), donor DNA produced transformants of which up to 30% carried a defective xylE gene after growth at 37 degrees C. Of recombinant clones that failed to express xylE at 37 degrees C, about 10% expressed the gene when grown at 22 degrees C,

关键词：不动杆菌属/遗传学不动杆菌属/代谢氨基酸序列碱基序列邻苯二酚2 3-双加氧酶染色体细菌双加氧酶类卡那霉素抗药性/遗传学分子序列数据诱变插入氧合酶类/遗传学质粒/遗传学聚合酶链反应/方法假单胞菌恶臭/遗传学序列分析 DNA 转化细菌

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Conversion of unsaturated fatty acids by bacteria isolated from compost

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CURRENT MICROBIOLOGY 1999年第4期38卷 250-255页

作者： Kaneshiro, T Kuo, TM Nakamura, LK USDA ARS Natl Ctr Agr Utilizat Res Peoria IL 61604 USA

A compost mixture amended with soybean oil was enriched in microorganisms that transformed unsaturated fatty acids (UFAs). When oleic acid or 10-ketostearic acid was the selective fatty acid, Sphingobacterium thalpophilum (NRRL B-23206, NRRL B-23208, NRRL B-23209, NRRL B-23210, NRRL B-23211, NRRL B-23212), Acinetobacter spp. (NRRL B-23207, NRRL B-23213), and Enterobacter cloacae (NRRL B-23264, NRRL B-23265, NRRL B-23266) represented isolates that produced either hydroxystearic acid, ketostearic acid, or incomplete decarboxylations. When ricinoleic (12-hydroxy-9-octadecenoic) acid was the selective UFA, Enterobacter cloacae (NRRL B-23257, NRRL B-23267) and Escherichia sp. (NRRL B-23259) produced 12-C and 14-C homologous compounds, and Pseudomonas aeruginosa (NRRL B-23256, NRRL B-23260) converted ricinoleate to a trihydroxyoctadecenoate product. Also, various Enterobacter, Pseudomonas, and Serratia spp. appeared to decarboxylate linoleate substrate incompletely. These saprophytic, compost bacteria were aerobic or facultative anaerobic Gram-negative and decomposed UFAs through decarboxylation, hydroxylation, and hydroperoxidation mechanisms.

关键词：不动杆菌属/代谢脂肪酸类不饱和/代谢革兰氏阴性兼性厌氧杆菌/代谢粪肥/微生物学假单胞菌属/代谢蓖麻油酸类/代谢土壤微生物学豆油/代谢

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INCIDENCE AND IDENTIFICATION OF PHOSPHOLIPASE C-PRODUCING BACTERIA IN FRESH AND SPOILED HOMOGENIZED MILK

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JOURNAL OF DAIRY SCIENCE 1976年第11期59卷 1857-1864页

作者： FOX, CW CHRISOPE, GL MARSHALL, RT UNIV MISSOURI DEPT FOOD SCI & NUTR COLUMBIA MO 65201 USA

Bacteria which produced phospholipase C were isolated from 13 of 34 fresh and 15 of 35 spoiled samples of homogenized milk. No single off flavor was assigned consistently to samples with phospholipase producers, but 75% of them were bitter. Pseudomonads constituted 62% of the isolates. Other phospholipase C-producing genera and their numbers were 2 Acinetobacter, 3 Alcaligenes, 2 Bacillus, 1 Citrobacter, 3 Enterobacter and 2 Flavobacterium. Two unidentified yeasts were isolated.

关键词：不动杆菌属/代谢产碱菌属/代谢芽孢杆菌属/代谢细菌/分离和提纯细菌/代谢柠檬酸杆菌属/代谢肠杆菌科/代谢黄杆菌属/代谢乳/微生物学乳/标准磷脂酶类/生物合成假单胞菌属/代谢动物

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