Studies on phosphate uptake by Acinetobacter calcoaceticus under aerobic conditions

Acinetobacter calcoaceticus was cultivated in a well-aerated stirred tank reactor and its phosphate uptake capacity was investigated. Statistical media optimization was done to figure out favourable growth conditions of Acinetobacter calcoaceticus NRRLB-552. Plackett–Burman design was used to figure out the key nutrients (sodium acetate, ammonium chloride and calcium chloride) featuring high growth and/or uptake of phosphate. The optimal concentrations for these nutrients were (sodium acetate 5.0 g/l, ammonium chloride 0.67 g/l, calcium chloride 0.05 g/l) obtained by central composite design (CCD) protocols and verified in shake flask cultivations. Predicted and experimental dry cell weights obtained using the optimized media were 2.046 and 2.54 g/l indicating 97% agreement. The optimal values of pH and temperature for growth and phosphate uptake were found to be 7.69 and 31.86 °C, respectively, using CCD. Batch kinetics was also established in shake flask and fermenter using optimized medium and environmental conditions. Phosphate uptakes of 21 mg/g biomass and 36 mg/g biomass were obtained in shake flask and fermenter, respectively. The possible inhibition of nutrients (carbon, nitrogen and phosphate) was also established under shake flask cultivation conditions. Growth of the bacteria was inhibited at a concentration higher than 0.4% carbon and 0.6% nitrogen. However increasing concentration of phosphate did not show any inhibitory effect on growth. The above kinetics and inhibition data will serve as suitable database for the development of a mathematical model for growth and its use will be able to facilitate appropriate reactor design for the removal of phosphates from industrial effluents.     

Diversion of the metabolic flux from pyruvate dehydrogenase to pyruvate oxidase decreases oxidative stress during glucose metabolism in nongrowing Escherichia coli cells incubated under aerobic, phosphate starvation conditions

Ongoing aerobic metabolism in nongrowing cells may generate oxidative stress. It is shown here that the levels of thiobarbituric acid-reactive substances (TBARSs), which measure fragmentation products of oxidized molecules, increased strongly at the onset of starvation for phosphate (P(i)). This increase in TBARS levels required the activity of the histone-like nucleoid-structuring (H-NS) protein. TBARS levels weakly increased further in DeltaahpCF mutants deficient in alkyl hydroperoxide reductase (AHP) activity during prolonged metabolism of glucose to acetate. Inactivation of pyruvate oxidase (PoxB) activity decreased the production of acetate by half and significantly increased the production of TBARS. Overall, these data suggest that during incubation under aerobic, P(i) starvation conditions, metabolic flux is diverted from the pyruvate dehydrogenase (PDH) complex (NAD dependent) to PoxB (NAD independent). This shift may decrease the production of NADH and in turn the adventitious production of H(2)O(2) by NADH dehydrogenase in the respiratory chain. The residual low levels of H(2)O(2) produced during prolonged incubation can be scavenged efficiently by AHP. However, high levels of H(2)O(2) may be reached transiently at the onset of stationary phase, primarily because H-NS may delay the metabolic shift from PDH to PoxB.     

草酸青霉YTY及其突变体解磷能力、pH和分泌有机酸的关系

本研究利用离子束-UV复合诱变草酸青霉YTY选育高效解磷突变体,分析了出发菌株YTY及其突变体解磷过程中的解磷能力、pH和有机酸的变化及其相关性,探讨草酸青霉的解磷机理。结果表明: 离子束-UV复合诱变选育获得5株高效解磷突变株P9-8、P9-9、P15-4、P15-6和P15-7,解磷能力均较YTY提高60%以上。解磷过程中突变株解磷能力及解磷速率均高于YTY,而pH显著低于YTY,同时,突变体分泌有机酸种类及含量发生了不同程度的变化,突变体和YTY均可分泌乳酸、乙酸和草酸,P9-8还产生了柠檬酸。皮尔逊相关分析显示,YTY及5株突变株的解磷能力和pH值呈显著负相关;YTY及其突变体(P15-4除外)的解磷能力与有机酸浓度和pH呈显著相关。分泌有机酸和降低环境pH值可能是草酸青霉解磷的内在机制,离子束-UV复合诱变可引起草酸青霉YTY有机酸分泌种类和分泌量的变化,还可能诱发YTY启动其他H⁺释放途径降低pH参与解磷。本研究为高效解磷青霉的开发及青霉解磷机理阐明提供了生物材料及理论依据。     

Ethanol and glycerol production under aerobic conditions by wild-type,respiratory-deficient mutants and a fusion product of Saccharomyces cerevisiae

Summary Experiments were performed to investigate growth, ethanol and glycerol production by wild-type strains (RHO) and respiratory-deficient (rho) mutants of Saccharomyces cerevisiae. Furthermore protoplasts were fused in order to enhance the fermentation capacity of a flocculent strain. At high substrate conditions, 150 g/l of saccharose, there is no difference in cell growth. However, at a glucose concentration of 10–20 g/l the mutants grow much slower. After 3 days of incubation at 28° C in a complete medium the viability of the two strains is the same. In minimal medium on the other hand the number of viable cells of the mutant is 100-fold reduced. All mutants tested showed a higher specific activity of alcohol dehydrogenase (ADH I) and an enhanced production of glycerol compared with the wild-type strain. By protoplast fusion a modified flocculent strain was obtained with higher specific activity of ADH I and a reduced biosynthesis of glycerol. However, the yields of ethanol (75–78%) are about the same for the wild-type strain and the rho mutants under aerobic conditions in absence of catabolite repression.     

Decomposition of humic acids by incubation in a soil water-extract under various conditions of oxygen availability

Summary Decomposition of humic acids suspended in a soil water-extract under various conditions of oxygen availability (aerobic, anaerobic, alternating aerobic/anaerobic) led to the formation of fulvic acids, lower molecular weight compounds, and humin, a more complex mixture of insoluble compounds. The transformation of humic acids and formation of new humic compounds were higher under aerobic conditions than under either anaerobic or alternating aerobic/anaerobic ones.Although subjected to decomposition under greatly differing conditions of oxygen availability, the residual humic acids showed essentially the same types of chemical alterations, an increase of functional groups containing oxygen and a decrease of hydrolysable ternary and quarternary fractions.     

萃取条件对TBP络合萃取混合有机酸的影响

采用磷酸三丁酯(TBP)络合萃取来自城市污泥厌氧发酵的混合有机酸.通过萃取条件优化,探讨利用TBP络合萃取混合有机酸的可能性.研究表明,在萃取剂TBP浓度为3.63 mol/L,混合有机酸浓度为20 g/L,萃取时间为5 min,萃取温度为35℃,油水相比为1:1,经过3次萃取之后,混合有机酸萃取效率可达到98.47%...     

磷胁迫条件下落叶松幼苗对难溶性磷的利用

以ＡｌＰＯ４为Ｐ源在温室内采用砂培的方法,研究了落叶松（Ｌａｒｉｘ ｇｍｅｌｉｎｉ）２年生幼苗对难溶性Ｐ酸盐的利用状况。结果表明,落叶松幼苗可以利用一定数量的ＡｌＰＯ４。在供ＡｌＰＯ４不接种菌根菌时,落叶松幼苗吸收的Ｐ可达正常供Ｐ时的３５．１％和６４．９％。不同菌种对落叶松幼苗利用ＡｌＰＯ４的影响不同。接种点柄乳牛肝菌（Ｓｕｉｌｌｕｓ ｇｒａｎｕｌａｔｕｓ）时,落叶松幼苗对ＡｌＰＯ４的利用量高于不     

Biodegradation of p-aminoazobenzene by Bacillus subtilis under aerobic conditions

Biological oxidation of organic dyes is important for textile industry wastewater treatment. The aim of this work was to assess the biodegradation kinetics of a specific azo-dye, p-aminoazobenzene. The degradation of p-aminoazobenzene by Bacillus subtilis was examined through batch experiments in order to investigate the effect of p-aminoazobenzene on the bacterial growth rate and elucidate the mechanism of dye degradation. The results proved that B. subtilis cometabolizes p-aminoazobenzene in the presence of glucose as carbon source, producing aniline and p-phenylenediamine as the nitrogen–nitrogen double bond is broken. The azo-dye was found to act as an inhibitor to microbial growth. A mathematical model was developed that describes cellular growth, glucose utilization, p-aminoazobenzene degradation and product formation. Received 26 July 1996/ Accepted in revised form 14 May 1997     

Variation of the adhesion to polystyrene of phenotypic mutants of Pseudomonas aeruginosa ATCC 27853 during starvation conditions

The aim of this work was to analyse the effects of different growth conditions (phosphate and contemporary carbon-phosphate starvation) on polystyrene adhesion of a strain of Pseudomonas aeruginosa ATCC 27853 and its four phenotypic mutants during experimental growth in starvation conditions. Bacterial adhesion was measured at 20, 40, 60 and 720 min. Data obtained showed that growth conditions are an important factor for the capacity of initial adhesion to inanimate surfaces. The analyses of adhesion of two phenotypic mutants (Mut-P-01 and Mut-P-02) isolated during growth on phosphate starvation is interesting. This kind of experiment yields important information on the prevention of nosocomial infections.     

Microbial degradation of hydrocarbons in soil during aerobic/anaerobic changes and under purely aerobic conditions

Microbial hydrocarbon degradation in soil was studied during periodical aerobic/anaerobic switching and under purely aerobic conditions by using a pilot-scale plant with diesel-fuel-contaminated sand. The system worked according to the percolation principle with controlled circulation of process water and aeration. Periodical switching between 4 h of aerobic and 2 h of anaerobic conditions was achieved by repeated saturation of the soil with water. Whatever the cultivation mode, less than 50% of the diesel was degraded after 650 h because the hydrocarbons were adsorbed. Contrary to expectations, aerobic/anaerobic changes neither accelerated the rate of degradation nor reduced the residual hydrocarbon content of the soil. Obviously the pollutant degradation rate was determined mainly by transport phenomena and less by the efficiency of microbial metabolism. The total mass of oxygen consumed and carbon dioxide produced was greater under aerobic/anaerobic changing than under aerobic conditions, although the mass of hydrocarbons degraded was nearly the same. As shown by an overall balance of microbial growth and by a carbon balance, the growth yield coefficient was smaller during aerobic/anaerobic changes than under aerobic conditions. Received: 25 November 1997 /  Received revision: 15 January 1998 / Accepted: 18 January 1998     

Characterization and variations of organic parameters in teleost fish endolymph during day-night cycle, starvation and stress conditions

The aim of the present work was to examine the modifications of the organic composition of fish endolymph under environmental conditions (day-night cycle, starvation and Cl2-stress) known to modify otolith growth. Endolymph electrophoretic patterns were compared. An antibody raised against the trout otolith organic matrix allowed examining the variations of organic matrix precursors in the endolymph under the above conditions. Western blot analysis showed bands around 60-80 kDa. A 50% decrease of immunolabelling was observed during the night whereas increases were seen after starvation (factor 3) or stress (factor 2) suggesting that these variations could be related to the organic matrix deposit. A factor retarding in vitro CaCO3 crystallization (FRC) was shown to co-precipitate with endolymph proteins and its apparent molecular mass (determined by measuring the activity after electro elution of gel electrophoresis) was estimated around 20 kDa. The FRC activity was stable during day-night cycle whereas it decreased by 70% and nearly 100% under starvation and stress respectively. These results suggest that the FRC, although retarding in vitro crystallization, plays a major role in the process of otolith calcification and that the decreases measured after starvation and stress are responsible for the decreases of the otolith growth. The variations of these two parameters (precursors and FRC) could contribute for the changes in the microstructure of the otolith.     

Blood amino acid composition of poikilothermic vertebrate under prolonged starvation

Free amino acids in the blood plasma of lamprey (Lampetra fluviatilis) and frog (Rana temporaria) have been determined quantitatively for the periods of deprivation of the exogenous feed. The lamprey's total amino acid pool increased by 74% from November to April and reached the lower limit known for the mammals. The amount of free amino acids in frogs decreased by 40% in the spring as compared with the autumn values. The difference is accounted for by certain features of the living cycles of these animals. A more energetic proteolysis in the lamprey tissues as compared with that in the frog tissues has been confirmed by quantitative determining of leucine, isoleucine and valine in the blood of these animals. Apart from the above, alanine, glycine, lysine, threonine and, in certain periods, tyrosine have been found to be quantitatively significant in the plasma of both animal species. The composition and proportion of the amino acids in blood plasma of these animals are due to specific features of their metabolism and connected with the energy state of the liver cells under starvation.     

不同腐熟程度有机物料对土壤微生物群落功能多样性的影响

室内培养条件下,施用有机物料初期土壤微生物群落代谢功能Shannon多样性指数降低,中期又提高。有机物料种类和腐熟水平可明显影响土壤微生物群落对Biolog微平板中碳源的利用能力,土壤微生物群落利用各类碳源的能力随培养试验的延长而降低,在25d内新鲜有机物处理对碳源的利用率的下降速度低于同类腐熟有机物料处理。糖类是各处理土壤微生物群落的主要利用碳源。土壤微生物群落主成分分析表明,在施用有机物料后25d内腐熟水平是影响土壤微生物群落的主要因素,新鲜有机物处理的土壤微生物群落相似,腐熟有机物处理的土壤微生物群落相似,培养50d后各处理的土壤微生物群落无差异。     

Metabolism of 2,4,6-trinitrotoluene by aPseudomonas consortium under aerobic conditions

An aerobic bacterial consortium was shown to degrade 2,4,6-trinitrotoluene (TNT). At an initial concentration of 100 ppm, 100% of the TNT was transformed to intermediates in 108 h. Radiolabeling studies indicated that 8% of [¹⁴C]TNT was used as biomass and 3.1% of [¹⁴C]TNT was mineralized. The first intermediates observed were 4-amino-2,6-dinitrotoluene and its isomer 2-amino-4,6-dinitrotoluene. Prolonged incubation revealed signs of ring cleavage. Succinate or another substrate—e.g., malic acid, acetate, citrate, molasses, sucrose, or glucose—must be added to the culture medium for the degradation of TNT. The bacterial consortium was composed of variousPseudomonas spp. The results suggest that the degradation of TNT is accomplished by co-metabolism and that succinate serves as the carbon and energy source for the growth of the consortium. The results also suggest that this soil bacterial consortium may be useful for the decontamination of environmental sites contaminated with TNT.     

Production of phenylalanine and organic acids by phosphoenolpyruvate carboxylase-deficient mutants ofEscherichia coli

Summary Isogenic strains ofEscherichia coli were grown aerobically in minimal medium in a 2-liter airlift fermentor to determine whether appc (phosphoenolpyruvate carboxylase) mutation had the effect of directing glucose carbon into phenylalanine synthesis. Two host strains, YMC9 (ppc ⁺) and KB285 (ppc ^–) were used, either with (Phe^c) or without (Phe⁰) a plasmid which determines constitutive phenylalanine production. Carbon consumption and metabolic products were monitored. Phenylalanine production occurred only in strains carrying the Phe^c plasmid.ppc ^– strains produced less cell mass and more acetate, pyruvate, and phenylalanine (in the Phe^c strains) than did isogenicppc ⁺ strains. Lactate and ethanol production were not detected in any of the strains. Phe^c strains produced less acetate and pyruvate than their Phe⁰ homologs. Importantly,ppc ^–/Phe^c produced at least six times as much phenylalanine (0.32 g phenylalanine/g dry weight cells) asppc ⁺/Phe^c. Even in this case, however, phenylalanine was produced at ten-fold lower levels than acetate. Thus, although theppc ^– mutation stimulates phenylalanine production, it also stimulates the production of unwanted by-products such as acetate and pyruvate.