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Dental root canal treatment is required when dental caries progress to infection of the dental pulp. A major goal of this treatment is to provide complete decontamination of the dental root canal system. However, the morphology of dental root canal systems is complex, and many human dental roots have inaccessible areas. In addition, dental reinfection is fairly common. In conventional treatment, a cotton pellet and paper point made from plant cellulose is used to dry and sterilize the dental root canal. Such sterilization requires a treatment material with high absorbency to remove any residue, the ability to improve the efficacy of intracanal medication and high biocompatibility. Bacterial cellulose (BC) is produced by certain strains of bacteria. In this study, we developed BC in a pointed form and evaluated its applicability as a novel material for dental canal treatment with regard to solution absorption, expansion, tensile strength, drug release and biocompatibility. We found that BC has excellent material and biological characteristics compared with conventional materials, such as paper points (plant cellulose). BC showed noticeably higher absorption and expansion than paper points, and maintained a high tensile strength even when wet. The cumulative release of a model drug was significantly greater from BC than from paper points, and BC showed greater compatibility than paper points. Taken together, BC has great potential for use in dental root canal treatment.

In the beer brewing industry, microbial spoilage presents a consistent threat that must be monitored and controlled to ensure the palatability of a finished product. Many of the predominant beer spoilage microbes have been identified and characterized, but the mechanisms of contamination and persistence remain an open area of study. Post-production, many beers are distributed as kegs that are attached to draft delivery systems in retail settings where ample opportunities for microbial spoilage are present. As such, restaurants and bars can experience substantial costs and downtime for cleaning when beer draft lines become heavily contaminated. Spoilage monitoring on the retail side of the beer industry is often overlooked, yet this arena may represent one of the largest threats to the profitability of a beer if its flavor profile becomes substantially distorted. In this study, we sampled and cultured microbial communities found in beers dispensed from a retail draft system to identify the contaminating bacteria and yeasts. We also evaluated their capability to establish new biofilms in a controlled setting. Among four tested beer types, we identified over a hundred different contaminant bacteria and nearly twenty wild yeasts. The culturing experiments demonstrated that most of these microbes were viable and capable of joining new biofilm communities. From these data, we provide an important starting point for the efficient monitoring of beer spoilage in draft systems and provide suggestions for cleaning protocol improvements that can benefit the retail community.ImportanceBeer production, packaging, and service are each vulnerable to contamination by microbes that metabolize beer chemicals and impart undesirable flavors, which can result in the disposal of entire batches. Therefore, great effort is taken by brewmasters to reduce and monitor contamination during production and packaging. A commonly overlooked quality control stage of a beer supply chain is at the retail service end, where beer kegs supply draft lines in bars and restaurants under non-sterile conditions. We found that retail draft line contamination is rampant and that routine line cleaning methods are insufficient to efficiently suppress beer spoilage. Thus, many customers unknowingly experience spoiled versions of the beers they consume. This study identified the bacteria and yeast that were resident in draft beer samples and also assessed their abilities to colonize tubing material as members of stable biofilm communities.

Resumen Introducción. Las levaduras, las bacterias ácido lácticas y ácido acéticas, involucradas en la fermentación de cacao, son las responsables del consumo de los compuestos en la pulpa para desarrollar los precursores de sabor dentro del grano de cacao. Sin embargo, la fermentación es difícil de controlar por la complejidad de este proceso y las múltiples variables que la afectan, entre ellas la genética del cultivo. Objetivo. Analizar las transformaciones bioquímicas de diferentes grupos genéticos del cacao (Theobroma cacao L.) durante una micro fermentación controlada. Materiales y métodos. Se estableció una metodología de micro fermentación, con un cultivo iniciador de H. opuntiae y Acetobacter spp. en 4 grupos genéticos, obtenidos de clones comerciales del CATIE e híbridos de fincas de Upala. A lo largo de la fermentación se analizaron glucosa, fructosa, etanol, ácido láctico y acético en pulpa y metilxantinas (teobromina y cafeína) y (-)-epicatequina en grano. Resultados. No se encontraron diferencias significativas en el contenido y comportamiento de azúcares, ácidos orgánicos o etanol en pulpa a lo largo de la fermentación, pero sí en el tiempo de fermentación. Se observó una disminución de metilxantinas y epicatequinas, debido a reacciones enzimáticas y no enzimáticas. El contenido promedio de epicatequina resultó ser el compuesto distintivo entre los grupos genéticos del CATIE y cafeína en los cacaos comerciales. Conclusión. La metodología empleada posee un efecto regulador en el comportamiento de algunos metabolitos estudiados en pulpa de diferentes genéticas a lo largo de la fermentación, así como epicatequina y metilxantinas en grano. Abstract Introduction. Yeasts, lactic acid and acetic acid bacteria, involved in cocoa fermentation are responsible for consuming the compounds in the pulp to develop flavor precursors within the cocoa bean. However, cocoa fermentation is difficult to control due to the complexity of this process and the multiple variables that affect it, including the genetics of the crop. Objective. To analyze the biochemical transformations of different genetic groups of cocoa (Theobroma cacao L.) during a controlled micro-fermentation. Materials and methods. A micro-fermentation methodology was developed with a starter culture of H. opuntiae and Acetobacter spp. in 4 genetic groups, grown in the CATIE Clonal Garden and hybrids from different farms in the Upala area. Throughout the fermentation process glucose, fructose, ethanol, lactic acid and acetic acid were analyzed in pulp, and methylxanthines (theobromine and caffeine) and (-)-epicatechin in the bean. Results. No significant differences were found in the content and dynamics of sugars, organic acids and ethanol in pulp throughout the fermentation, but differences were found at fermentation time. A decrease in methylxanthines and epicatechin triggered by enzymatic and non-enzymatic reactions was observed. The average epicatechin content proved to be the distinctive compound between CATIE's genetic groups and caffeine in commercial cocoa. Conclusion. The methodology used had a regulatory effect on the behavior of some of the studied metabolites in pulp of different genetic throughout fermentation, as well as epicatechin and methylxanthines in the bean.

Many black water or feces and urine field observations in Tanjung Pinang City were dumped directly into the ditches, rivers, and sea. This proves that the management of domestic liquid waste in the City of Tanjungpinang, especially black water, has not yet received serious attention, resulting in a decline in environmental quality. One of the stages in processing black water is decomposition. The decomposition process can be assisted by bacteria, one of which is cellulose-degrading bacteria. This study aims to find bacterial isolates that have the potential to degrade cellulose in feces from coastal soils in Tanjung Pinang City. This research is a laboratory experimental research. The results of this study were obtained 16 bacterial isolates that can degrade cellulose. From these 16 isolates, 3 genera of bacteria were obtained, namely Pseudomonas, Acetobacter and Acinetobacter. Keywords: Cellulose Degradation Bacteria, Coastal Soil

In this research batch reactors were operated with coffee processing waste and autochthonous microbial consortium, and a taxonomic and functional analysis was performed for phase I of stabilization of maximum H2 production and for phase II of maximum H2 consumption. During phase I, the reactor's operating conditions were pH 4.84 to 8.18, headspace 33.18% to 66.82%, and pulp and husk from 6.95 to 17.05 g/L. These assays continued for phase II, with initial pH conditions of 5.8-8.1, headspace of 33.18-66.82%, and pulp and husk remaining from phase I. The highest homoacetogenesis was observed in assay 5 with pH 7.7, 40% headspace, and 15 g/L of pulp and husk (initial concentrations of phase I). A relative abundance of Clostridium 41%, Lactobacillus 20% and Acetobacter 14% was observed in phase I. In phase II, there was a change in relative abundance of 21%, 63%, and 1%, respectively, and functional genes involved with autotrophic (formyltetrahydrofolate synthase) and heterotrophic (enolase) homoacetogenesis, butanol (3-hydroxybutyryl-CoA dehydrogenase), and propionic acid (propionate CoA-transferase) were identified. This study provides a new and amplified insight into the physicochemical and microbiological factors, which can be used to propose adequate operational conditions to maximize the bioenergy production and reduce homoacetogenesis in biological reactors.

4 Tablas.-- 4 Figuras, A new aeration treatment of black ripe olive streams prior to their vacuum evaporation was studied. It was found that the aeration for two days of these solutions reduced the chemical oxygen demand (COD) of the distillate phase (>80%) generated during their vacuum evaporation; although the temperature (14–38 °C) and the air flow rate (10–50 L air/h/L) had a great influence on this reduction. Moreover, the COD in the distillate was mainly associated with the presence of acetic acid (~400 mg/L) and ethanol (~500 mg/L), the later substance being eliminated by strong aeration. Additionally, the growth of native aerobic microorganisms, mainly Acetobacter, increased the pH of the wastewater to 7–8 units, thus avoiding evaporation of the acetic acid salt formed. These results make the use of the distillate for plant irrigation viable and its discharge into municipal depuration plants possible, given the low COD (, This work was supported by the Spanish Government (Project AGL2016-76820-R, AEI/FEDER, UE).

Microbial contamination of fuels, associated with a wide variety of bacteria and fungi, leads to decreased product quality and can compromise equipment performance by biofouling and microbiologically influenced corrosion of pipelines and storage tanks. Detection and quantification of biomass are critical in monitoring fuel systems for an early detection of microbial outbreaks. The aims of this study are (i) to quantify bacterial and fungal contamination in samples from diesel storage tanks of petrol stations, using both culture dependent- and culture independent (qPCR) approaches, and (ii) to analyse the diversity of cultivable diesel-contaminating microorganisms with the purpose to create a strain collection for further use in biodeterioration experiments. Both methodological approaches revealed a high microbial contamination in all studied samples, with the bacterial load being much higher than the fungal load. The diversity of cultivable microorganisms was rather low. Based on criteria of abundance and fuel degradation potential, the most relevant microorganisms were identified as bacteria of genera Bacillus , Citrobacter , Burkholderia and Acetobacter , the filamentous fungi Paecilomyces variotii and Pseudallescheria boydii , and a Dipodascaceae yeast. Furthermore the validity and utility of qPCR-based methods are discussed.

Bacterial cellulose (BC) is a naturally produced as an exopolysaccharide from some bacteria. It has excellent properties over the plant cellulose and has numerous applications in many fields including food, pharmaceutical, textile, paper manufacturing and other industries. However, a major limitation of bacterial cellulose production is the high cost of carbon substrate. The study aims to reduce the cost of bacterial cellulose using a cheap carbon source. This study presents feasibility in the production of bacterial cellulose using the starchy effluent waste from the Thai fermented rice noodle manufacturing process as a low-cost substrate by Acetobacter xylinum TISTR 975. The optimizations of culture conditions for bacterial cellulose production were also investigated under static culture. The results indicated that starchy effluent waste from the Thai fermented rice noodle manufacturing process performs well for the production of bacterial cellulose by supplementing with 50 g/L sucrose and 2% olive oil under the static condition. The structure and physical properties of bacterial cellulose were characterized using SEM, FTIR and XRD. In summary, the starchy effluent waste from the Thai fermented rice noodle manufacturing process can be used to produce bacterial cellulose which is a high value-added, sustainable and green product.

Shake-flask cultures of microorganisms involve flame sterilization during sampling, which produces combustion gas with high CO2 concentrations. The gaseous destination has not been deeply analyzed. Our aim was to investigate the effect of flame sterilization on the headspace of the flask and on the shake-flask culture. In this study, the headspace CO2 concentration was found to increase during flame sterilization ~0.5–2.0% over 5–20 s empirically using the Circulation Direct Monitoring and Sampling System. This CO2 accumulation was confirmed theoretically using Computational Fluid Dynamics; it was 9% topically. To evaluate the influence of CO2 accumulation without interference from other sampling factors, the flask gas phase formed by flame sterilization was reproduced by aseptically supplying 99.8% CO2 into the headspace, without sampling. We developed a unit that can be sampled in situ without interruption of shaking, movement to a clean bench, opening of the culture-plug, and flame sterilization. We observed that the growth behaviour of Escherichia coli, Pelomonas saccharophila, Acetobacter pasteurianus, and Saccharomyces cerevisiae was different depending on the CO2 aeration conditions. These results are expected to contribute to improving microbial cell culture systems.

Physiological responses to changes in environmental conditions such as temperature may partly arise from the resident microbial community that integrates a wide range of bio-physiological aspects of the host. In the present study, we assessed the effect of developmental temperature on the thermal tolerance and microbial community of Drosophila melanogaster. We also developed a bacterial transplantation protocol in order to examine the possibility of reshaping the host bacterial composition and assessed its influence on the thermotolerance phenotype. We found that the temperature during development affected thermal tolerance and the microbial composition of male D. melanogaster. Flies that developed at low temperature (13°C) were the most cold resistant and showed the highest abundance of Wolbachia, while flies that developed at high temperature (31°C) were the most heat tolerant and had the highest abundance of Acetobacter. In addition, feeding newly eclosed flies with bacterial suspensions from intestines of flies developed at low temperatures changed the heat tolerance of recipient flies. However, we were not able to link this directly to a change in the host bacterial composition.

Background/Objectives: In India the increase in population and industrialisation, there is a much need for the soil to be used for various activities. There is a need to improve the properties of soil, if the soil at any particular locality is unsuited, wholly or partially selected for the construction. With increasing awareness of environmental issues, there has been a remarkable shift toward “green” and sustainable technologies. In view of these attempt is made to study the new sustainable method of biogeochemical techniques to improve the soil properties. Methods: In this work test for the Unconfined Compressive Strength (UCS) for soils were conducted as per IS to know variation in strength characteristics with the addition of two bacteria namely Acetobacter and Bacillus of different incubation periods and curing periods. For the same soil samples, the free swell index values were also determined to know the effect of addition of bacteria by stabilisation. Findings: The UCC test results for the soil samples treated with bacteria showed an increase in strength of soil with the increase in age of bacteria. The increase in Unconfined Compressive Strength (UCS) value at 60 day curing period is about 175% for soil treated with Acetobacter and 520% for Bacillus. Swelling characteristics also reduced by the treatment with Acetobacter and Bacillus bacteria. Improvement/Applications: Biostabilisation with bacteria proved the increase in strength and decrease in swell it can be used as an innovative stabiliser for stabilising problematic soils, which is cost-effective, non intrusive, low energy demanding and ecofriendly.

Some Acetobacter strains produce a cellulose called bacterial cellulose in culture medium. This cellulose has unique structural and mechanical properties compared with higher plant cellulose, and is expected to be a commodity material in various fields of industry. For economical mass production, it is essential to construct an aeration and agitation culture process. To explore the industrial applications of the bacterial cellulose, the structural features and physicochemical properties need to be understood and potentially improved. This paper reviews recent progress in research on this bacterial cellulose.

Methods of adapting micro-organisms to an inhibiting factor in an active industrial bioprocess were examined with an acetic acid fermentation as model. With the aim of automatic control, a fuzzy-logic system was developed on the basis of the collected knowledge of skilled vinegar brewers. In a first step, this fuzzy system was to assess the actual adaptation degree of the bacteria on the basis of data from robust and reasonably priced sensors. From this information an appropriate setpoint value for the inhibiting factor ‘final acid concentration’ was derived for each batch cycle. As a result a further acid tolerance was found after several batch cycles. This adaptation effect should be used to increase the product concentration to more than 20 g per 100 ml acetic acid with a high productivity. The stepwise adapted culture was productive over the aimed acetic acid concentration, a 10% improvement of the product formation rate could be found compared with the status before conditioning. High product concentration and increased productivity finally result in shorter cycle times, less transport and storage volumes, an improved utilization of energy and material resources, and, last but not least, they are an essential steps towards the fulfillment of economical and ecological demands.

Aims To maximize acetification rate (ETA) by adsorption of acetic acid bacteria (AAB) on loofa sponge matrices (LSM). Methods and Results AAB were adsorbed on LSM, and the optimal shaking rate was determined for maximized AAB growth and oxygen availability. Results confirm that the 1 Hz reciprocating shaking rate with 40% working volume (liquid volume 24 l, tank volume 60 l) achieved a high oxygen transfer coefficient (kLa). The highest ETA was obtained at 50% (w:v) LSM-AAB:culture medium at 30 ± 2°C (P ≤ 0·05). To test process consistency, nine sequential acetification cycles were run using LSM-AAB and comparing it with no LSM. The highest ETA (1·701–2·401 g l−1 d−1) was with LSM-AAB and was associated with the highest biomass of AAB, confirmed by SEM images. Conclusions Results confirm that LSM-AAB works well as an inert substrate for AAB. High oxygenation was maintained by a reciprocating shaker. Both shaking and LSM were important in increasing ETA. Significance and Impact of the Study High cell biomass in LSM-AAB provides good conditions for higher ETAs of quick acetification under adequate oxygen transfer by reciprocating shaker. It is a sustainable process for small-scale vinegar production system requiring minimal set-up cost.

The fermentation of cocoa pulp is one of the few remaining large-scale spontaneous microbial processes in today's food industry. The microbiota involved in cocoa pulp fermentations is complex and variable, which leads to inconsistent production efficiency and cocoa quality. Despite intensive research in the field, a detailed and comprehensive analysis of the microbiota is still lacking, especially for the expanding Asian production region. Here, we report a large-scale, comprehensive analysis of four spontaneous Malaysian cocoa pulp fermentations across two time points in the harvest season and two fermentation methods. Our results show that the cocoa microbiota consists of a "core" and a "variable" part. The bacterial populations show a remarkable consistency, with only two dominant species, Lactobacillus fermentum and Acetobacter pasteurianus. The fungal diversity is much larger, with four dominant species occurring in all fermentations ("core" yeasts), and a large number of yeasts that only occur in lower numbers and specific fermentations ("variable" yeasts). Despite this diversity, a clear pattern emerges, with early dominance of apiculate yeasts and late dominance of Saccharomyces cerevisiae. Our results provide new insights into the microbial diversity in Malaysian cocoa pulp fermentations and pave the way for the selection of starter cultures to increase efficiency and consistency. ispartof: PLOS ONE vol:8 issue:12 ispartof: location:United States status: published

The coproduction of a useful material and electricity via a novel application of microbial fuel cell (MFC) technology to oxidative fermentation was investigated. We focused on vinegar production, i.e., acetic acid fermentation, as an initial and model useful material that can be produced by oxidative fermentation in combination with MFC technology. The coproduction of acetic acid and electricity by applying MFC technology was successfully demonstrated by the simultaneous progress of acetic acid fermentation and electricity generation through a series of repeated batch fermentations. Although the production rate of acetic acid was very small, it increased with the number of repeated batch fermentations that were conducted. We obtained nearly identical (73.1%) or larger (89.9%) acetic acid yields than that typically achieved by aerated fermentation (75.8%). The open-cycle voltages measured before and after fermentation increased with the total fermentation time and reached a maximum value of 0.521 V prior to the third batch fermentation. The maximum current and power densities measured in this study (19.1 μA/cm2 and 2.47 μW/cm2, respectively) were obtained after the second batch fermentation.

The modeling and optimization of industrial processes requires an intensive study of the factors involved. In this work, a continuous pilot system for studying the industrial process of acetic fermentation is developed. A Doehlert design is applied to the five variables involved in the pilot process. This experimental design allows reduction of the experimental burden and the maximum amount of information to be obtained, studying the factors at different levels depending on their significance. The experimental system provides a robust measure of the specific growth rate and the rates of substrates consumption and acetic acid production, related to the flow of effluent stream evaluated in the steady state. The results demonstrate the growth-associated kinetics of substrates and product, and the yield factors are calculated with low values of variances for the coefficients, i.e., within the range 1-11%. The specific growth rate suits the quadratic model proposed. The response surfaces generated by the model are applied to explain the behavior of the bacterial growth and, therefore, the effects of the process variables studied over the acetic acid production. Very low levels of ethanol or oxygen make the acetification rate decrease, and a saturation effect with high levels of ethanol or oxygen is also deduced. The effects of the aeration rate, agitation, and overpressure suggest a kind of inhibition of the acetic acid production caused by the oxygen that has not been practically studied before. The temperature strengthens the inhibitory effect of the ethanol and the oxygen. The conclusions of this work consolidate the structure of a hybrid model for the acetic fermentation.

Cell-free extracts of Acetobacter suboxydans were prepared which were capable of condensing α-ketoisovalerate with 14 C-labeled acetyl-coenzyme A to yield 14 C-labeled α-isopropylmalate. The product of the reaction was isolated by paper and column chromatography and was characterized by recrystallization with synthetic α-isopropylmalic acid to constant specific radioactivity. The formation of α-isopropylmalate by extracts of A. suboxydans plus the ability of the organism to grow in a simple glucose-glycerol medium containing glutamic acid as the only amino acid indicate that the pathway for leucine biosynthesis shown to exist in yeast and Salmonella typhimurium also occurs in A. suboxydans . As a comparison, the condensation of oxalacetate and ( 14 C) acetyl-coenzyme A to yield ( 14 C) citric acid was shown, by similar means, to occur in A. suboxydans . This is of interest since the existence of this classical condensing enzyme has hitherto not been demonstrated in this organism. This reaction was further demonstrated in cell-free extracts of A. suboxydans by means of a spectrophotometric assay at 232 mμ which measured the cleavage of the carbon-sulfur bond of acetyl-coenzyme A in the presence of oxalacetate. Comparison of the specific activities of crude cell-free extracts indicated a much more extensive occurrence of this reaction in yeast than in A. suboxydans .

Dihydroxyacetone was quantitatively produced from glycerol during the primary exponential growth phase and depleted during the secondary exponential phase. Although no growth was detected on the basal medium, growth occurred upon addition of dihydroxyacetone.

d-Glucosaminic acid has recently been found to be an oxidized product of d-glucosamine formed by Ps. fluorescens. It has been revealed that many strains of oxidative bacteria can oxidize glucosamine. The formation of glucosamine acid has been recognized among a large number of strains of Pseudomonas, Acetobacter and Gluconobacter, by means of paper chromatography. Furthermore, one of these strains, A. melanogenum Beijerinck, oxidized glucosamine to glucosaminic acid with the theoretical consumption of oxygen as Ps. fluorescens liquefaciens. Glucosaminic acid was proved by isolation and identification by means of using resting cells.The experiment of growth shows that Ps. fluorescens liq. could not secure any energy by means of the oxidation of glucosamine.

D-Glucosaminic acid has recently been found to be an oxidized product of D-glucosamine formed by Ps. fluorescens. It has been revealed that many strains of oxidative bacteria can oxidize glucosamine. The formation of glucosaminic acid has been recognized among a large number of strains of Pseudomonas, Acetobacter and Gluconobacter, by means of paper chromatography. Furthermore, one of these strains, A. melanogenum Beijerinck, oxidized glucosamine to glucosaminic acid with the theoretical consumption of oxygen as Ps. fluorescens liquefaciens. Glucosaminic acid was proved by isolation and identification by means of using resting cells. The experiment of growth shows that Ps. fluorescens liq. could not secure any energy by means of the oxidation of glucosamine.

Summary: Of 11 strains of Acetobacter tested, three strains of A. xylinum (IFO 3288, IFO 13693 and IFO 13772), two strains of A. aceti (IFO 3281 and IFO 3283), one strain of A. pasteurianus (IFO 3223) and one strain of A. rancens (IFO 3297) were found to produce 2-25 mg per 100 ml culture filtrate of a mixture of (1→2)-β-D-glucans and (1→2)-β-D-gluco-oligosaccharides as low-molecular-weight products. The glucans and oligosaccharides were linear without branching structures and their degrees of polymerization (DPs) ranged from 6 to about 42. The compounds with lower DPs did not seem to be produced by hydrolysis of those with higher DPs. A. xylinum IFO 3288 and A. xylinum IFO 13693 also produced extracellular acidic polysaccharides containing L-rhamnose, d-glucose, d-mannose and uronic acid.

Preliminary results obtained during the study of vinegar fermentation show that the conversion yield can be increased to 100% of theoretical and the acetic acid productivity can reach 7g/l.h., in continuous culture without cell immobilisation.

1. Growing cells of a small number of strains of Acetomonas oxydans oxidized d-quinate to 5-dehydroquinate. 2. d-Shikimate was oxidized to 4,5-dihydroxy-3-oxocyclohex-1-ene-1-carboxylate (3-dehydroshikimate, formerly 5-dehydroshikimate). 3. d-Dihydroshikimate was oxidized to the corresponding 5-dehydro compound, but epidihydroshikimate oxidation by growing cells was not observed. 4. Cell-free extracts oxidized d-quinate to 5-dehydroquinate with the consumption of the stoicheiometric amount of oxygen, but oxidation of shikimate and dihydroshikimate did not go to completion. 5. Oxidation of quinate was brought about by a constitutive particulate enzyme probably localized in the cytoplasmic membrane. No evidence was found for the participation of NAD, NADP or free flavine compounds in electron transport, but the system was cytochrome-linked.

Acetobacter suboxydans does not contain an active tricarboxylic acid cycle, yet two pathways have been suggested for glutamate synthesis from acetate catalyzed by cell extracts: a partial tricarboxylic acid cycle following an initial condensation of oxalacetate and acetyl coenzyme A. and the citramalate-mesaconate pathway following an initial condensation of pyruvate and acetyl coenzyme A. To determine which pathway functions in growing cells, acetate- 1 - 14 C was added to a culture growing in minimal medium. After growth had ceased, cells were recovered and fractionated. Radioactive glutamate was isolated from the cellular protein fraction, and the position of the radioactive label was determined. Decarboxylation of the C5 carbon removed 100% of the radioactivity found in the purified glutamate fraction. These experiments establish that growing cells synthesize glutamate via a partial tricarboxylic acid cycle. Aspartate isolated from these hydrolysates was not radioactive, thus providing further evidence for the lack of a complete tricarboxylic acid cycle. When cell extracts were analyzed, activity of all tricarboxylic acid cycle enzymes, except succinate dehydrogenase, was demonstrated.