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Improvements in lipase production and recovery from Acinetobacter radioresistens in presence of polypropylene powders filled with carbon sources.

Polypropylene powders as the adsorbent for organic solution containing n-hexadecane and olive oil were employed as the carbon source for producing an alkaline lipase from Acinetobacter radioresistens. The best volumetric ratio of n-hexadecane to olive oil around 5 for lipase production was determined from shake-flask and fermentation cultivations. The existence of a maximum time course lipase activity of the aqueous phase was attributed to the compensation effects of olive oil on cell growth and lipase production, repression of lipase synthesis by oleic acid, and lipase adsorption on the supports. A linear relationship between the average cell growth rate in the exponential phase and the ratio of surface areas of the supports was found. The benefits of using the present fermentation process include less foaming and emulsion of the broth, less organic phase used, higher lipase production, and easy recovery of the lipase in the centrifugation step.

Acinetobacter↗

A comparison of sole carbon source utilization patterns and phospholipid fatty acid profiles to detect changes in the root microflora of hydroponically grown crops.

Sole carbon source utilization (SCSU) patterns and phospholipid fatty acid (PLFA) profiles were compared with respect to their potential to characterize root-inhabiting microbial communities of hydroponically grown crops. Sweet pepper (Capsicum annum cv. Evident), lettuce (Lactuca sativa cv. Grand Rapids), and four different cultivars of tomato (Lycopersicon esculentum cvs. Gitana, Armada, Aromata, and Elin) were grown in 1-L black plastic beakers placed in a cultivation chamber with artificial light. In addition to the harvest of the plants after 6 weeks, plants of one tomato cultivar, cv. Gitana, were also harvested after 4 and 8 weeks. The cultivation in this study was performed twice. Principal component analysis was used to analyze the data. Both characterization methods had the ability to discriminate between the root microflora of different plant species, cultivars, and one tomato cultivar at different ages. Differences in both SCSU patterns and PLFA profiles were larger between plant species than between cultivars, but for both methods the largest differences were between the two cultivations. Still, the differences between treatments were always due to differences in the same PLFAs in both cultivations. This was not the case for the SCSU patterns when different plant ages were studied. Furthermore, PLFA profiles showed less variation between replicates than did SCSU patterns. This larger variation observed among the SCSU data indicates that PLFA may be more useful to detect changes in the root microflora of hydroponically grown crops than the SCSU technique.

Capsicum↗

Metabolic fluxes in Corynebacterium glutamicum during lysine production with sucrose as carbon source.

Metabolic fluxes in the central metabolism were determined for lysine-producing Corynebacterium glutamicum ATCC 21526 with sucrose as a carbon source, providing an insight into molasses-based industrial production processes with this organism. For this purpose, 13C metabolic flux analysis with parallel studies on [1-(13C)Fru]sucrose, [1-(13C)Glc]sucrose, and [13C6Fru]sucrose was carried out. C. glutamicum directed 27.4% of sucrose toward extracellular lysine. The strain exhibited a relatively high flux of 55.7% (normalized to an uptake flux of hexose units of 100%) through the pentose phosphate pathway (PPP). The glucose monomer of sucrose was completely channeled into the PPP. After transient efflux, the fructose residue was mainly taken up by the fructose-specific phosphotransferase system (PTS) and entered glycolysis at the level of fructose-1,6-bisphosphate. Glucose-6-phosphate isomerase operated in the gluconeogenetic direction from fructose-6-phosphate to glucose-6-phosphate and supplied additional carbon (7.2%) from the fructose part of the substrate toward the PPP. This involved supply of fructose-6-phosphate from the fructose part of sucrose either by PTS(Man) or by fructose-1,6-bisphosphatase. C. glutamicum further exhibited a high tricarboxylic acid (TCA) cycle flux of 78.2%. Isocitrate dehydrogenase therefore significantly contributed to the total NADPH supply of 190%. The demands for lysine (110%) and anabolism (32%) were lower than the supply, resulting in an apparent NADPH excess. The high TCA cycle flux and the significant secretion of dihydroxyacetone and glycerol display interesting targets to be approached by genetic engineers for optimization of the strain investigated.

Carbon Isotopes↗

Endo-polygalacturonase in Saccharomyces wine yeasts: effect of carbon source on enzyme production.

Eight wine yeast strains of Saccharomyces sp. were tested for polygalacturonase (PGase) activity, after cultivation on various carbon sources. No strain showed any activity when grown on glucose, while five strains produced PGase in the presence of galactose and polygalacturonate. These data suggest that the PGase of wine strains is repressed by glucose and induced by galactose and polygalacturonate. The existence of the PGase gene in the wine strains and its similarity with that of the laboratory strains was proved by Southern hybridization and PCR amplification. The promoter region of the PGase gene in the wine strains was slightly different from that of the laboratory strains. This possibly explains the different pattern of gene expression in wine and laboratory strains. The PGase of wine strains produced di- or tri-galacturonic acid from polygalacturonic acid, different from the fungal PGase.

Carbon↗

Biological sulfate reduction using molasses as a carbon source.

The feasibility of using a laboratory-scale upflow anaerobic sludge blanket process for sulfate reduction with molasses as a carbon source was demonstrated. Competition between methane-producing bacteria (MPB) and sulfate-reducing bacteria (SRB) was influenced by the chemical oxygen demand-to-sulfur (COD:S) ratio in the feed. Sulfate removal greater than 80% could be achieved at COD:S greater than 10 when MPB predominated. Activity of MPB and SRB was inhibited at a dissolved sulfide concentration of approximately 200 mg/L. Competition between MPB and SRB was intense as the COD:S was reduced from 5 to 2. Further reduction in the COD:S to 0.7 led to the formation of sulfidogenic granules. The COD removal decreased to approximately 30% at a COD:S less than 2 because of accumulation of sulfurous precipitates and the nonbiodegradable portion of molasses in the sludge. Reduced gas production rates further imposed limitations on diffusion of the organic substrate into granules. Sulfidogenic process operation yielded sulfate removal as great as 70% at a COD:S of approximately 3.5.

Bacteria, Anaerobic↗

[Effect of the carbon source and aeration conditions on homoserine lysine biosynthesis in the threonine-dependent mutant Brevibacterium flavum 2T].

The effect of two carbon sources (sucrose and acetate), aeration conditions and threonine concentration on the homoserine and lysine biosynthesis by the threonine-dependent mutant Brevibacterium flavum 2T was examined. It was demonstrated that acetate provided the predominant synthesis of homoserine to a far greater extent than sucrose (with the weight/weight ratio of homoserine : lysine being 2.5-5.0 and 0.8-1,2, respectively). The maximal level of homoserine and lysine was 18-21 and 3-7 g/l on the acetate containing medium and 18-22 and 12-16 g/l on the sucrose containing medium, respectively. On sucrose the total amount of amino acids and the total yield of products as related to the consumed substrate were greater than on acetate. Using the sucrose medium, the effect of aeration conditions and threonine concentration on the biosynthesis of both compounds was investigated. With an aeration increase from 1.3 to 4.6 g O2/l.hr the optimal concentration of threonine in the medium grow. The biosynthesis of homoserine was less sensitive to the inhibitory effect of excessive threonine than that of lysine. With an increase of the threonine concentration in the medium from 0.25 to 3.0 g/l the ratio homoserine : lysine grew from 1.03 to 5.20 (with the sulphite number being 4.6 g O2/l.hr). This effect was independent of the aeration conditions.

Aerobiosis↗

Biological nutrient removal by a sequencing batch reactor (SBR) using an internal organic carbon source in digested piggery wastewater.

Experiments in a lab-scale SBR were conducted to demonstrate the feasibility of using an internal carbon source (non-digested pig manure) for biological nitrogen and phosphorus removal in digested piggery wastewater. The internal C-source used for denitrification had similar effects to acetate. 99.8% of nitrogen and 97.8% of phosphate were removed in the SBR, from an initial content in the feed of 900 mg/l ammonia and 90 mg/l phosphate.

Animals↗

Regulation of peroxisomal proteins and organelle proliferation by multiple carbon sources in the methylotrophic yeast, Candida boidinii.

A methylotrophic yeast, Candida boidinii, was grown on various combinations of peroxisome-inducing carbon source(s) (PIC(s)), i.e. methanol, oleate and D-alanine, and the regulation of peroxisomal proteins (both matrix and membrane ones) and organelle proliferation were studied. This regulation was followed (1) at the protein or enzyme level by means of the peroxisomal enzyme activity and Western analysis; (2) at the mRNA level by Northern analysis; and (3) at the organelle level by direct observation of peroxisomes under a fluorescent microscope. Peroxisomal proliferation was followed in vivo by using a C. boidinii strain producing a green fluorescent protein having peroxisomal targeting signal 1. When multiple PICs were used for cell growth, C. boidinii induced specific peroxisomal proteins characteristic of all PIC(s) present in the medium, responding to all PIC(s) simultaneously. Thus, these PICs were considered to induce peroxisomal proliferation independently and not to repress peroxisomes induced by other PICs. Next, the sensitivity of the peroxisomal induction to glucose repression was studied. While the peroxisomal induction by methanol or oleate was completely repressed by glucose, the D-alanine-induced activities of D-amino acid oxidase and catalase, Pmp47, and the organelle proliferation were not. These results indicate that peroxisomal proliferation in yeasts is not necessarily sensitive to glucose repression. Lastly, this regulation was shown to occur at the mRNA level.

Acyl-CoA Oxidase↗

Production of proteinase on noncarbohydrate carbon sources by Pseudomonas aeruginosa.

Proteinase production by Pseudomonas aeruginosa was studied in medium containing noncarbohydrate materials, especially various hydrocarbons, as the sole carbon source. On heavy oil, kerosene, n-paraffinic hydrocarbon of C(12), C(14), or C(16), and propylene glycol, the bacteria grew well and high protinase production was observed. However, production on paraffinic hydrocarbon differed remarkably with strains of varied origins. The elastase-positive strain, IFO 3455, showed abundant growth and high proteinase production on medium containing a paraffin of C(12), C(14), or C(16), whereas the elastase-negative strain, IFO 3080, showed little growth on the same medium. Neither elastase-positive nor elastase-negative strains, however, utilized n-paraffins of C(5) to C(10), or various aromatic hydrocarbons such as benzene, naphthalene, phenanthrene, and anthracene. The proteinases produced on the noncarbohydrate medium were identical with those produced in glucose medium.

Culture Media↗

A physiological evaluation of carbon sources for calcification in the octocoral Leptogorgia virgulata (Lamarck).

The union of calcium cations with carbonate anions to form calcium carbonate (CaCO3) is a fundamentally important physiological process of many marine invertebrates, in particular the corals. In an effort to understand the sources and processes of carbon uptake and subsequent deposition as calcium carbonate, a series of studies of the incorporation of 14C-labeled compounds into spicules was undertaken using the soft coral Leptogorgia virgulata. It has been surmised for some time that dissolved inorganic carbon in sea water is used in the biomineralization process. Furthermore, it was suspected that metabolically generated CO2 is also available for calcification. As a means of testing these possible sources of carbon in spicule calcification, key enzymes or transport systems in each pathway were inhibited. First, the enzyme carbonic anhydrase was specifically inhibited using acetazolamide. Second, the active transport of bicarbonate was inhibited using DIDS (4,4'-diisothiocyanato-stilbene-2,2'-disulfonic acid). Third, CO2 generation resulting from glycolysis and the citric acid cycle was arrested using iodoacetic acid, which interferes specifically with the enzyme glyceraldehyde-3-phosphate dehydrogenase. The results indicate that dissolved CO2 is the largest source of carbon used in the formation of calcitic sclerites, followed by HCO3- from dissolved inorganic carbon. In L. virgulata, the dissolved inorganic carbon is responsible for approximately 67% of the carbon in the sclerites. The other 33% comes from CO2 generated by glycolysis. Two important conclusions can be drawn from this work. First, carbon for spiculogenesis comes not only from dissolved inorganic carbon in the environment but also from metabolically produced carbon dioxide. While the latter has been theorized, it has never before been demonstrated in octocorals. Second, regardless of the carbon source, the enzyme carbonic anhydrase plays a pivotal role in the physiology of spicule formation in Leptogorgia virgulata.

Animals↗

Bioemulsifier production in batch culture using glucose as carbon source by Candida lipolytica.

The yeast Candida lipolytica IA 1055 produced an inducible extracellular emulsification activity while utilizing glucose at different concentrations as carbon source during batch fermentation at 27 degrees C. In all glucose concentrations studied, maximum production of emulsification activity was detected in the stationary phase of growth, after pH reached minimal values. The bioemulsifier isolated was a complex biopolymer constituting proteins, carbohydrates, and lipids. The results obtained in this work show that the biosynthesis of a bioemulsifier is not simply a prerequisite for the degradation of extracellular hydrocarbon.

Biomass↗

CO2 as main carbon source for isoprenoid biosynthesis via the mevalonate-independent methylerythritol 4-phosphate route in the marine diatoms Phaeodactylum tricornutum and Nitzschia ovalis.

Isoprenoid biosynthesis was investigated in the two diatoms Phaeodactylum tricornutum and Nitzschia ovalis by labeling experiments performed in mixotrophic growth conditions with sodium [1-(13)C]acetate, 13CO2, [1-(13)C]glucose, sodium [3-(13)C]pyruvate and 1-deoxy-D-[5,5-(2)H2]xylulose. A clear dichotomy was found. Acetate was the preferred carbon source for the formation of the sterols in the cytoplasm via the mevalonate pathway. Carbon dioxide was the main source for phytol biosynthesis in the chloroplasts via the mevalonate-independent methylerythritol 4-phosphate pathway. The two diatoms showed the same compartmentation for isoprenoid biosynthesis as that previously found in higher plants, the red alga Porphyridium cruentum and the Chrysophyte Ochromonas danica.

Carbon Dioxide↗

The influence of mineral and carbon sources on biological control of charcoal rot fungus, Macrophomina phaseolina by fluorescent pseudomonads in tomato.

AIMS: To determine the influence of various trace minerals and carbon source on the biocontrol performance of Pseudomonas aeruginosa strain IE-6S+ and P. fluorescens strain CHA0 against Macrophomina phaseolina. METHODS AND RESULTS: In dual culture plate assay, P. aeruginosa IE-6S+ and P. fluorescens CHA0 inhibited radial growth of M. phaseolina producing zones of inhibition. Czapek's dox agar medium amended with both zinc and glucose remarkably improved antifungal activities of the bacterial inoculants. Under glasshouse conditions, soil amendment with zinc and/or glucose alone did not reduce M. phaseolina infection in tomato roots but did reduce significantly when used in combination with IE-6S+ or CHA0. Soil amendments with zinc and/or glucose increased fresh shoot weights but zinc amendment greatly reduced bacterial populations in the rhizosphere. CONCLUSIONS: Mineral and carbon amendments enhance the biocontrol performance of fluorescent pseudomonads against M. phaseolina. SIGNIFICANCE AND IMPACT OF THE STUDY: Identification of mineral and carbon amendments that favour biocontrol of certain bacterial strains may provide clues to soil factors or components of nutrient solutions in hydroponic culture that will improve the level and reliability of control.

Antibiosis↗

Differentiation of Rhizomucor species by carbon source utilization and isoenzyme analysis.

Nineteen Rhizomucor miehei and Rhizomucor pusillus isolates were assayed for their ability to utilize 87 various substrates as a single carbon source. Besides a difference in sucrose utilization, distinctive differences were found in the utilization of glycine, phenylalanine, and beta-alanine. Five isoenzyme systems also proved useful for the determination of markers of distinctive value at a species level. Data were used to obtain information about the genetic polymorphism of these species: a high degree of variability was found among the R. pusillus isolates, whereas the group of R. miehei isolates was more homogeneous genetically.

Animals↗

Anaerobic biodegradability of Tween surfactants used as a carbon source for the microbial reductive dechlorination of hexachlorobenzene.

Three structurally-related, nonionic, polysorbate surfactants (Tween 60, 61, and 65) were used as the sole carbon source to sustain the microbial, sequential reductive dechlorination of hexachlorobenzene (HCB) in a mixed, methanogenic culture derived from a contaminated estuarine sediment. The surfactants were partially degraded and fermented to methane with no measurable accumulation of volatile fatty acids, indicating that methanogenesis was rapid relative to the rates of hydrolysis and acidogenesis. Addition of the methanogenesis inhibitor 2-bromoethanesulfonic acid resulted in acetate accumulation without impact on the sequential dechlorination of HCB. An anaerobic biodegradability assay was performed and the following data were obtained for the Tween 60, 61, and 65, respectively: 53, 62, and 62% COD destruction; 35, 57, and 48% COD to methane conversion; and 38, 38, and 45% COD to acetate conversion. These data suggest that the hydrophobic moiety (stearate) of the surfactants was preferentially degraded, most likely through beta-oxidation, to acetate and ultimately to methane and carbon dioxide. Between 38 and 47% of the initial surfactant COD remained after 46 d incubation, which most likely corresponds to the hydrophilic polyoxyethylene moiety. An anaerobic biodegradation pathway of the Tween surfactants is proposed.

Alkanesulfonic Acids↗

[Effect of the COD and total phosphorus concentration on biological phosphorus removal supplied with acetate as a sole carbon source].

The effect of the concentration of acetate and total phosphorus on biological phosphorus removal process in sequencing batch reactor fed with sodium acetate (as sole carbon source) was studied in this paper. The reason of the inefficiency of phosphorus removal by sludge fed with high concentration acetate solution was analyzed. The results indicated that when COD < 600 mg.L-1 the phosphorus removal increased with the increase of COD/TP; and this removal increased obviously when COD/TP < 50 but not when COD/TP > 50. The phosphorus removal descended at high acetate concentration (COD > 600 mg.L-1) and broke down when COD > 1000 mg.L-1. It is the transmission of high acetate concentration from the anaerobic stage to the aerobic stage that led to the proliferation of the filamentous microorganism and the expansion of sludge, then polyP bacteria was wash out, which resulted in the drop of phosphorus removal.

Acetates↗

Biosynthesis of branched-chain amino acids in yeast: effect of carbon source on leucine biosynthetic enzymes.

The three enzymes in the leucine biosynthetic pathway of yeast do not exhibit coordinate repression and derepression in response to the carbon source available in the culture medium. Growth in an acetate medium results in derepression of the first enzyme in the pathway, alpha-isopropylmalate synthase, and repression of the second two enzymes, alpha-isopropylmalate isomerase and beta-isopropylmalate dehydrogenase, relative to the levels found in glucose-grown cells. The role of endogenous leucine pools as a mediator of these differences was investigated. The leucine pools did not differ significantly between acetate-grown and glucose-grown cells. However, an elevated endogenous leucine pool, caused by exogenous leucine in the growth medium, did decrease the rate of decay of alpha-isopropylmalate synthase activity observed when acetate-grown cells were shifted to glucose. Evidence is provided suggesting that an elevated endogenous leucine pool may increase the in vivo stability of alpha-isopropylmalate synthase under several different conditions. Studies on the kinetics of alpha-isopropylmalate synthase decay in vivo and sensitivity to leucine inhibition indicate that there are two classes of the enzyme in acetate-grown yeast cells.

Acetates↗