Survival of lux-marked bacteria introduced into soil and the rhizosphere of bean (Phaseolus vulgaris L.)

The survival of lux-marked recombinants of Escherichia coli and Bacillus subtilis was studied in the rhizosphere of bean (Phaseolus vulgaris L.) and in bulk soil. The number of E. coli (pSB343) containing a complete lux operon did not differ significantly according to whether they were introduced into soil separately or together with a non-luminescent mutant Pseudomonas fluorescens R2fN. When genetically altered strains of E. coli and B. subtilis bearing a complete or an incomplete lux-reporter system were introduced into soil, the numbers of surviving cells were the same both in the rhizosphere and bulk soil. The insertion of lux genes into bacterial strains therefore does not affect their competitiveness and survival in the rhizosphere and bulk soil.The author is with the Department of Microbiology, University of Silesia, Jagielloska 28, 40-032 Katowice, Poland     

Effects of organic compounds on the degradation ofp-nitrophenol in lake and industrial wastewater by inoculated bacteria

Many microorganisms fail to degrade pollutants when introduced in different natural environments. This is a problem in selecting inocula for bioremediation of polluted sites. Thus, a study was conducted to determine the success of four inoculants to degradep-nitrophenol (PNP) in lake and industrial wastewater and the effects of organic compounds on the degradation of high and low concentrations of PNP in these environments.Corynebacterium strain Z4 when inoculated into the lake and wastewater samples containing 20 µg/ml of PNP degraded 90% of PNP in one day. Addition of 100 µg/ml of glucose as a second substrate did not enhance the degradation of PNP and the bacterium utilized the two substrates simultaneously. Glucose used at the same concentration (100 µg/ml), inhibited degradation of 20 µg of PNP in wastewater byPseudomonas strain MS. However, glucose increased the extent of degradation of PNP byPseudomonas strain GR. Phenol also enhanced the degradation of PNP in wastewater byPseudomonas strain GR, but had no effect on the degradation of PNP byCorynebacterium strain Z4.Addition of 100 µg/ml of glucose as a second substrate into the lake water samples containing low concentration of PNP (26 ng/ml) enhanced the degradation of PNP and the growth ofCorynebacterium strain Z4. In the presence of glucose, it grew from 2×10⁴ to 4×10⁴ cells/ml in 3 days and degraded 70% of PNP as compared to samples without glucose in which the bacterium declined in cell number from 2×10⁴ to 8×10³ cells/ml and degraded only 30% PNP. The results suggest that in inoculation to enhance biodegradation, depending on the inoculant, second organic substrate many play an important role in controlling the rate and extent of biodegradation of organic compounds.Abbreviations PNP p-nitrophenol     

Emission of volatile organic compounds by apple trees under spider mite attack and attraction of predatory mites

Emission rates of volatile organic compounds (VOCs) from Pirus malus L. subsp. mitis (Wallr.) var. Golden Delicious and var. Starking attacked by the phytophagous mite Panonychus ulmi Koch, and their attractiveness to the predatory mites Amblyseius andersoni Chant and Amblyseius californicus McGregor, were studied during three years. A large variability was found in the emission of individual VOCs depending on the infestation, the apple tree variety and the date. There were larger total VOC emission rates and larger total VOC leaf concentrations in apple trees attacked by phytophagous mites, especially in the var. Starking. In infested trees of this variety, there were also more predatory mites. An olfactometer assay showed that predatory mites preferentially chose branches infested by Panonychus ulmi (85% went to infested branches vs 15% to uninfested control branches) indicating that volatiles may be used as cues to find their prey.     

Metabolism of volatile phenolic compounds from hydroxycinnamic acids byBrettanomyces yeast

The formation of 4-ethyl and 4-vinyl derivatives of guaiacol, phenol and syringol from ferulic acid,p-coumaric acid and sinapic acid, respectively, byBrettanomyces sp. in a synthetic medium was studied by gas chromatography-mass spectrometry. Some of these metabolites possess strong spicy, smoke-like, medicinal, clove-like, woody or phenolic odours and their role as spoilage compounds in wine is discussed. Their formation appears to be characteristic of this yeast genus and its sporulating formDekkera, suggesting these yeasts are Pof⁺. This paper attempts to clarify the distinctive and characteristic odours which have long been attributed toBrettanomyces yeast metabolism.     

Variations in volatile compounds fromElshoizia cilliata

We used gas chromatography-mass spectrometry (GC-MS) to analyze total yields and relative composition of the volatile compounds from leaf extracts ofElshoizia cilliata. This species contains 22 compounds. The major constituents of its essential oils are dihydrotagetone (62.7%), ß-caryophellene (4.96%), germacrene-d (4.03%), and ±-humulene (1.34%). Compounds in these leaf extracts are remarkably high in dihydrotagetones (40.5 to 81.6%). The total amount of monoterpenoids is 3.17 to 7.03 times greater than that of sesquiterpenoids, and is highly correlated with the level of dihydrotagetone (r = 0.97). Seasonal variations are significantly different only for dihydrotagetone (p < 0.0005), but not for the yields of other volatile compounds.

     

樟树叶片挥发性有机物释放季节动态和日动态变化规律

采用动态顶空采集法和热脱附-气质联用技术,对不同季节和一天内不同时间点樟树叶片释放的挥发性有机物(VOCs)成分含量及其变化规律进行了分析。结果表明:全年樟树叶片释放的VOCs共计78种,其中萜烯类(19种)和烷烃类(18种)化合物种类较多; 32种樟树叶片释放的挥发性成分相对含量在不同季节存在显著差异(P<0.05); 3月份释放的VOCs种类最为丰富,萜烯类化合物相对含量最高(43.49%),主要为1-石竹烯、D-柠檬烯和α-蒎烯,其他月份释放的VOCs则以烷烃类和醇类为主;春季,樟树叶片在绝大多数时间点释放VOCs以萜烯类化合物为主,且早上8:00释放的萜烯类化合物相对含量最大。综上结果认为,樟树是营造保健型生态园林的理想树种,其叶片释放的VOCs中富含多种对人体健康有益的萜烯类化合物,春季的早晨是进行森林康养活动的最佳时间。     

Distribution and immunological characterization of microbial aldehyde reductases

The distribution of microbial aldo-keto reductases was examined and their immunochemical characterization was performed. p-Nitrobenzaldehyde, pyridine-3-aldehyde and ethyl 4-chloro-3-oxobutanoate reductase activities were found to be widely distributed in a variety of microorganisms. In immunodiffusion studies, most yeasts belonging to the genera Sporobolomyces, Sporidiobolus and Rhodotorula formed precipitin bands with anti-Sporobolomyces salmonicolor aldehyde reductase serum. Furthermore, the results of immunotitration experiments suggested that Sporobolomyces salmonicolor AKU 4429 contains other enzyme(s) which can reduce p-nitrobenzaldehyde, pyridine-3-aldehyde and/or ethyl 4-chloro-3-oxobutanoate, and which are inactivated by anti-Sporobolomyces salmonicolor aldehyde reductase serum.     

Effects of nonmethane volatile organic compounds on microbial community of methanotrophic biofilter

Effects of nonmethane volatile organic compounds (NMVOCs) on methanotrophic biofilter were investigated. Laboratory-scale biofilters packed with pumice and granular-activated carbon (10:1, w/w) were operated with CH₄ and NMVOCs including dimethyl sulfide (DMS) and benzene/toluene (B/T). DMS alone exhibited a positive effect on the methanotrophic performance; however, the coexistence of B/T removed this effect. B/T alone exerted no effect on the performance. Pyrosequencing and quantitative PCR revealed that the NMVOCs strongly influenced the bacterial and methanotrophic communities but not the population density of methanotrophs. DMS alone diversified and changed both bacterial and methantrophic communities, but its effects were nullified by the presence of B/T. Canonical correspondence analysis revealed significant correlations between the NMVOCs and community composition and significant interaction between DMS and B/T. DMS did not affect the distribution of types I/II methanotrophs (60/40), while B/T increased the abundance of type I to 82 %. DMS and B/T favored the growth of the methanotrophic bacteria Methylosarcina and Methylomonas, respectively. These results suggest that NMVOCs can be a significant abiotic factor influencing methane biofiltration.     

三酮类除草剂微生物降解的研究进展

三酮类除草剂是一类高效、广谱、高选择性的除草剂,能够有效地防除玉米地多种阔叶杂草和禾本科杂草。该类除草剂在土壤、水体中残留,易造成地表和地下水污染。有关研究表明,微生物降解有望成为解决该类除草剂残留的有效措施。本文分析了2种三酮类除草剂带来的生态效应,总结了已报道的微生物降解资源,简述了降解基因/酶的研究进展及可能的降解途径,为深入研究三酮类除草剂的生物降解提供一定的信息支撑。     

Production of volatile aroma compounds by bacterial strains isolated from different surface-ripened French cheeses

Twelve bacterial strains belonging to eight taxonomic groups: Brevibacterium linens, Microbacterium foliorum, Arthrobacter arilaitensis, Staphylococcus cohnii, Staphylococcus equorum, Brachybacterium sp., Proteus vulgaris and Psychrobacter sp., isolated from different surface-ripened French cheeses, were investigated for their abilities to generate volatile aroma compounds. Out of 104 volatile compounds, 54 volatile compounds (identified using dynamic headspace technique coupled with gas chromatography-mass spectrometry [GC-MS]) appeared to be produced by the different bacteria on a casamino acid medium. Four out of eight species used in this study: B. linens, M. foliorum, P. vulgaris and Psychrobacter sp. showed a high flavouring potential. Among these four bacterial species, P. vulgaris had the greatest capacity to produce not only the widest varieties but also the highest quantities of volatile compounds having low olfactive thresholds such as sulphur compounds. Branched aldehydes, alcohols and esters were produced in large amounts by P. vulgaris and Psychrobacter sp. showing their capacity to breakdown the branched amino acids. This investigation shows that some common but rarely mentioned bacteria present on the surface of ripened cheeses could play a major role in cheese flavour formation and could be used to produce cheese flavours.     

Analysis of volatile compounds released from embryogenic cultures and somatic embryos of sweet oranges by head space SPME

Volatile compounds released from callus and nucellar embryo tissues of ‘Valencia late’ and ‘Washington Navel’ sweet oranges (Citrus sinensis, L. Osbeck) were collected/concentrated by head space solid phase micro extraction and analysed by gas chromatography-mass spectrometry. Friable, white embryogenic cultures released a number of volatile compounds, including some essential oils. Different samples of the same embryogenic culture showed variability, possibly related to the presence of tissues undergoing differentiation. Analyses of the somatic embryos permitted the identification of several components, including limonene and methyl anthranilate. Considering the simplicity and the very small sample required (0.3 g of fresh tissue) head space solid phase micro extraction is suitable for studies and comparisons of volatile metabolites released from in vitro Citrus tissue cultures suggesting its potential in Citrus biochemical, genetic and breeding research. This revised version was published online in June 2006 with corrections to the Cover Date.     

固相微萃取-气相色谱-质谱结合电子鼻技术分析发酵乳中挥发性风味物质

【背景】德氏乳杆菌保加利亚亚种(Lactobacillus delbrueckii subsp. bulgaricus)和嗜热链球菌(Streptococcus thermophilus)作为酸奶的常用发酵剂,在乳制品生产中的应用十分广泛。【目的】在前期研究基础上,以筛选出的具有良好风味的复配发酵剂(复配比为1:100)L.delbrueckiisubsp.bulgaricusIMAU20312与S.thermophilusIMAU80809为实验对象,分析牛乳在发酵及贮藏期间风味物质的动态变化规律,为发酵剂的开发与应用提供依据。【方法】采用固相微萃取-气相色谱-质谱联用(solid-phasemicroextraction-gaschromatography-mass spectrometry, SPME-GC-MS)结合电子鼻技术检测并解析新鲜牛乳在发酵及贮藏阶段所产生的挥发性风味物质。【结果】该发酵剂在牛乳发酵及贮藏期间产生酸类物质23种,醛类物质13种,醇类物质15种,酮类物质18种,酯类物质11种,烷烃类物质22种。香气活度值(odor activity value, OAV...     

A novel method for characterisation of microbial growth kinetics on volatile organic compounds

A novel method for the determination of microbial growth kinetics on hydrophobic volatile organic compounds (VOC) has been developed. A stirred tank reactor was operated as a fed-batch system to which the VOC was continuously fed via the gas phase, assuring a constant VOC concentration in the mineral medium. A flow of air was saturated with the VOC, and then mixed with a further flow of air, to obtain a predetermined VOC concentration. Thus, different VOC concentrations in the mineral medium could be obtained by altering the VOC concentration in the feed gas. The growth kinetics of Xanthobacter autotrophicus GJ10 on 1,2-dichloroethane (DCE) and of Pseudomonas sp. strain JS150 on MonoChloroBenzene (MCB) were assessed using this method. The growth of strain JS150 was strongly inhibited at MCB concentrations higher than 160 mg l⁻¹, and the results were fitted using a piecewise function. The growth kinetics of strain GJ10 were described by the Luong model where maximum growth rate μ_max = 0.12 h⁻¹, substrate saturation constant K _S = 7.8 mg l⁻¹, and maximum substrate concentration S _m (above which growth is completely inhibited) = 1080 mg l⁻¹. Varying nitrogen and oxygen flows enabled the effect of oxygen concentration on the growth kinetics of Pseudomonas JS150 to be determined. Received: 30 November 1998 / Received revision: 19 March 1999 / Accepted: 20 March 1999     

Kinetics of Light Emission and Oxygen Consumption by Bioluminescent Bacteria

Oxygen plays a key role in bacterial bioluminescence. The simultaneous and continuous kinetics of oxygen consumption and light emission during a complete exhaustion of the exogenous oxygen present in a closed system has been investigated. The kinetics are performed with Vibrio fischeri, V. harveyi, and Photobacterium phosphoreum incubated on respiratory substrates chosen for their different reducing power. The general patterns of the luminescence time courses are different among species but not among substrates. During steady-state conditions, substrates, which are less reduced than glycerol, have, paradoxally, a better luminescence efficiency. Oxygen consumption by luciferase has been evaluated to be 17% of the total respiration. Luciferase is a regulatory enzyme presenting a positive cooperative effect with oxygen and its affinity for this final electron acceptor is about 4–5 times higher than the one of cytochrome oxidase. The apparent Michaelis constant for luciferase has been evaluated to be in the range of 20 to 65 nM O₂. When O₂ concentrations are as low as 10 nM, luminescence can still be detected; this means that above this concentration, strict anaerobiosis does not exist. By n-butyl malonate titration, it was clearly shown that electrons enter the luciferase pathway only when the cytochrome pathway is saturated. It is suggested that, in bioluminescent bacteria, luciferase acts as a free-energy dissipating valve when anabolic processes (biomass production) are impaired.     

Laccase-catalyzed oxidation of phenolic compounds in organic media

Rhus vernificera laccase-catalyzed oxidation of phenolic compounds, i.e., (+)-catechin, (−)-epicatechin and catechol, was carried out in selected organic solvents to search for the favorable reaction medium. The investigation on reaction parameters showed that optimal laccase activity was obtained in hexane at 30 °C, pH 7.75 for the oxidation of (+)-catechin as well as for (−)-epicatechin, and in toluene at 35 °C, pH 7.25 for the oxidation of catechol. E_a and Q₁₀ values of the biocatalysis in the reaction media of the larger log p solvents like isooctane and hexane were relatively higher than those in the reaction media of lower log p solvents like toluene and dichloromethane. Maximum laccase activity in the organic media was found with 6.5% of buffer as co-solvent. A wider range of 0–28 μg protein/ml in hexane than that of 0–16.7 μg protein/ml in aqueous medium was observed for the linear increasing conversion of (+)-catechin. The kinetic studies revealed that in the presence of isooctane, hexane, toluene and dichloromethane, the K_m values were 0.77, 0.97, 0.53 and 2.9 mmol/L for the substrate of (+)-catechin; 0.43, 0.34, 0.14 and 3.4 mmol/L for (−)-epicatechin; 2.9, 1.8, 0.61 and 1.1 mmol/L for catechol, respectively, while the corresponding V_max values were 2.1 × 10⁻², 2.3 × 10⁻², 0.65 × 10⁻² and 0.71 × 10⁻² δA/μg protein min); 1.8 × 10⁻², 0.88 × 10⁻², 0.19 × 10⁻² and 1.0 × 10⁻² δA/μg protein min); 0.48 × 10⁻², 0.59 × 10⁻², 0.67 × 10⁻² and 0.54 × 10⁻² δA/μg protein min), respectively. FT-IR indicated the formation of probable dimer from (+)-catechin in organic solvent. These results suggest that this laccase has higher catalytic oxidation capacity of phenolic compounds in suitable organic media and favorite oligomers could be obtained.     

Elicitation of volatile compounds in photomixotrophic cell culture of Petroselinum crispum

Photomixotrophic cells of Petroselinum crispum accumulated >500 mg chlorophyll per kg wet weight and grew well in a broad range of phytoeffector conditions. Autoclaved fungal cells were lethal for photoheterotrophic cells, but induced in photomixotrophic cells the formation of volatile n-alkanes, phthalides, coumarins, and elemicine. Most of the compounds elicited reached a concentration maximum between 20 and 30 h after addition of the mycelium, whereas the group of n-alkanes increased steadily during the 90 h monitored. Maximum concentrations were: 12 mg of graveolone, 1 mg of bergapten, 0.5 mg of sedanenolide, and 0.5 mg of n-tetradecane per 1 nutrient medium. A dose/effect relationship was found; 10 to 25 g of fungal wet weight per 1 culture medium resulted in maximum accumulation of volatiles. The formation of volatiles by photomixotrophic in vitro cells is discussed as an integral part of plant responses to ecological stress.     

Degradation of aromatic compounds byRhizobium spp.

Summary The ability of rhizobia to utilize catechol, protocatechuic acid, salicylic acid, p-hydroxybenzoic acid and catechin was investigated. The degradation pathway of p-hydroxybenzoate byRhizobium japonicum, R. phaseoli, R. leguminosarum, R. trifolii andRhizobium sp. isolated from bean was also studied.R. leguminosarum, R. phaseoli andR. trifolii metabolized p-hydroxybenzoate to protocatechuate which was cleaved by protocatechuate 3,4-dioxygenasevia ortho pathway.R. japonicum degraded p-hydroxybenzoate to catechol which was cleaved by catechol 1,2-dioxygenase.Rhizobium sp., a bean isolate, dissimilatedp-hydroxybenzoate to salicylate. Salicylate was converted to gentisic acid prior to ring cleavage. The rhizobia convertedp-hydroxybenzoate to Rothera positive substance. Catechol and protocatechuic acid were directly cleaved by the species.R. japonicum converted catechin to protocatechuic acid.     

Effect of cobalt ions on the metabolism of some volatile and polar compounds in the marine invertebrates Mytilus galloprovincialis and Actinia equina

The compositions of the volatile and polar fractions from two coexisting Black Sea invertebrates, the mussel Mytilus galloprovincialis and the beadlet anemone Actinia equina, were established. The main metabolites in the volatile fraction from the investigated animals appeared to be methyl esters of fatty acids and fatty aldehydes. In the polar fraction from both animals low concentrations of free acids and nitrogen-containing compounds were obtained. Free carbohydrates were in much higher concentrations in M. galloprovincialis than in A. equina. Some sterols, probably as polar conjugates, were identified mainly in A. equina. Significant changes among all compounds appeared after treatment of both invertebrates with two different concentrations of cobalt ions. The variety of changes in each invertebrate could be due to their different evolutionary status. The effect of cobalt ions was often stronger at medium cobalt-ion concentrations.