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Pseudomonas cepacia utilized penicillin G as the sole source of carbon and energy. We report here an unexplained correlation between lysine auxotrophy and beta-lactamase deficiency, resulting in loss of capacity to utilize penicillin.
Threonine can be used aerobically as the sole source of carbon and energy by mutants of Escherichia coli K-12. The pathway used involves the conversion of threonine via threonine dehydrogenase to aminoketobutyric acid, which is further metabolized by aminoketobutyric acid ligase, forming acetyl coenzyme A and glycine. A strain devoid of serine transhydroxymethylase uses this pathway and excretes glycine as a waste product. Aminoketobutyric acid ligase activity was demonstrated after passage of crude extracts through Sephadex G100.
The effect of biotin on the specific growth rate of Candida tropicalis was studied on media with various sources of carbon nutrition at temperatures of 29 and 39degreesC. The yeast was found to require 3--5 times less biotin on the medium with paraffin than on the medium with glucose and hexodecane, and 3--4 times more biotin on the same substrate with an increase in temperature. The growth rate increased by 36--40% on liquid paraffins containing 0.5 mcg/litre of biotin at 29degreesC and 1.5--2.0 mcg/litre of biotin at 39degreesC. The saturation constant of biotin increased from 0.033 to 0.063 mcg/litre on the medium with paraffin, and more by an order on the medium with glucose and hexadecane, at a rise of growth temperature from 29 to 39degreesC.
Some characteristic are presented for DNA of active sludge adapted to domestic waste, diethylene glycol, mannitol and glycerol. A significant shift towards CG-type is found for the nucleotide composition of the DNA of active sludge adapted to glycerol and mannitol. The fusion curves for DNA of the active sludge evidence for the presence of some totality of molecules in it. In case of the diethylene glycol adapted active sludge DNA fusion two transitions are observed corresponding to 37.8 and 68.2 mol.% of GC-pairs, that may be explained by bimodality in distribution of the number of species of the free-living bacteria according to the DNA nucleotide composition. The fusion curves for DNA of the active ooze adapted to domestic waste, mannitol and glycerol has no clearly pronounced transitions, that may be explained by the presence of other groups of microorganisms in these sludges, besides bacteria. Heterogeneity of DNA relative to the nucleotide composition with transition to a single source of carbon decreases in the series: initial active sludge greater than active sludge adapted to mannitol greater than glycerol greater than diethylene glycol.
Nonsulfur photosynthetic purple bacteria isolated from the Dzhermuk mineral springs (Armenia) were grown on sugar-containing media and found to be capable of synthesizing L(+)-lactic acid. Various organic compounds were tested as possible sole sources of carbon and an electron donors required to support bacterial growth and biosynthesis of lactic acid under various growth conditions.
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A mixed microbial culture assimilating 4-chloroaniline and 3,4-dichloroaniline as sole sources of carbon and nitrogen was isolated from soil treated with propanide for a long period of time. The process is accompanied with a 100% liberation of chloride ions. Aniline cannot serve as a growth substrate for the mixed culture. The authors have studied whether the mixed culture utilizing chloroanilines can decompose these compounds in model experiments with natural waters and soil suspensions. The culture actively decomposes 3,4-dichloroaniline (20 mg/litre) in natural water samples into which it has been added at a concentration of 10(3)--10(4) cells/ml. The decomposition is accelerated in experiments with suspensions of meadow-chernozem soils when cells of the mixed culture are added to the suspensions. The rate of 3,4-dichloroaniline degradation in a grey forest soil suspension remains nearly unchanged when the mixed culture is added to it. Degradation of the chloroaniline is accompanied with equivalent liberation of chloride ions when the mixed culture is added to natural water samples.
Various methods, including selection by resistance to the antibiotic mecillinam have been used to obtain Sh. flexneri mutants capable of utilizing mannitol and mannose as the sources of carbon. The mutation which determines this phenotype is localized in str-spc area and cotransduced with str A gene (28-33%). The appearance of this mutation in the initial Sh. flexneri strain 516 led to a decrease in the virulence of this strain for guinea-pigs. The restoration of its properties and the partial restoration of its virulence could be detected in Mal+ hybrids obtained by crossing with E. coli strain K12- the product of this gene may participate in the regulation of the synthesis of some pathogenicity factor.
The oleaginous yeast, Candida curvata D, was grown in both batch and continuous culture on 5 different carbon sources to compare the efficiency of fat production from the various substrates. Maximum lipid accumulation occurred in batch culture with xylose as the carbon source on nitrogen-limited medium reaching a level of 49% (w/w) of the biomass, but this was reduced to 37% at the optimum dilution rate (D = 0.05/hr) in a chemostat. Both the highest biomass and lipid yields were attained in continuous culture with lactose as the sole carbon source at a dilution rate of D = 0.04/hr, giving an efficiency of substrate conversion of 60 g of biomass and 18.6 g lipid per 100 g lactose utilized. The relative proportions of the major fatty acids (16:0, 18:0, 18:1, 18:2) in the lipid were found to vary considerably in batch culture and in continuous culture under carbon-limited conditions. However, on nitrogen-limited media in the chemostat, the fatty acid composition remained relatively constant over the whole range of dilution rates employed. Lipid from xylose-grown cells contained the greatest percentage of stearic acid (18:0) 15% and the lowest linoleic acid (18:2) 4%, whereas lipid from ethanol-grown cells contained elevated levels of oleic acid (18:1) 51% and decreased palmitic acid (16:0) 25%.
Escherichia coli K-12 has two acetohydroxy acid synthase (AHAS) isozymes (AHAS I and AHAS III). Both of these isozymes catalyze the synthesis of alpha-aceto-alpha-hydroxybutyrate and alpha-acetolactate, which are key intermediates of the isoleucine-valine biosynthetic pathway. Strains lacking either isozyme but not both activities have been previously shown to grow well in minimal media in the absence of isoleucine and valine on any of several commonly used carbon sources (e.g., glucose or succinate). We report the characterization of mutants that were unable to grow on either acetate or oleate as a sole carbon source due to a defect in isoleucine-valine biosynthesis. The defect in isoleucine-valine biosynthesis was expressed only on these carbon sources and was due to the loss of AHAS I activity, resulting from lesions in the ilvBN operon. Previously identified ilvBN mutant strains also failed to grow on acetate or oleate minimal media. Our results indicated that AHAS I is an essential enzyme for isoleucine and valine biosynthesis when E. coli K-12 is grown on acetate or oleate as the sole carbon source. AHAS III was expressed during growth on acetate or oleate but was somehow unable to produce sufficient amounts of alpha-aceto-alpha-hydroxybutyrate and alpha-acetolactate to allow growth.
Effects of fermentation conditions on the comonomer composition and its distribution of poly(3-hydroxybutyrate-co-3-hydroxypropionate) [P(3HB-co-3HP)] have been investigated for bacterial synthesis of P(3HB-co-3HP)s by Alcaligenes latus from sucrose and 3-hydroxypropionate (3HPA) mixed carbon sources. Comparison of the microstructures of these samples drew a conclusion that when the concentration of oxygen dissolved (DO) in the fermentation medium was controlled between 5 and 20% (based on the concentration at saturation), the 3HP content and the comonomer compositional distribution (CCD) of the copolymer would not be influenced by the DO values. The concentration of the carbon sources was monitored during the fermentation. The results indicated that the comonomer composition and its distribution of P(3HB-co-3HP)s were interrelated to the amounts of carbon sources transported into the bacterial cells. When the bacteria consumed more sucrose, the more 3HPA they would utilize, and the broader the CCD of the copolymer would be. Furthermore, the efficiencies of the transformation of the two carbon sources to the copolymer constituents were found to be similar.
The production of extracellular and cell-associated glucosyltransferase activity by Streptococcus mutans strain GS-5 was examined during growth on various carbon sources in a chemically defined medium. S. mutans cells produced glucosyltransferase activity only during logarithmic growth when glucose, fructose, mannitol, or sorbitol was the sole carbon source. Cells growing on mannitol or sorbitol produced approximately half as much extracellular glucosyltransferase activity as cells growing on glucose, although the proportions of the glucosyltransferase activity capable of synthesizing insoluble glucans were similar. Cells growing on fructose produced slightly more extracellular glucosyltransferase activity than cells grown on glucose, yet the proportion of the glucosyltransferase activity capable of synthesizing insoluble glucans was again similar to glucose cultures. S. mutans cells growing in the presence of both glucose and mannitol displayed diauxic growth and initial preferential utilization of glucose. Glucosyltransferase enzyme production occurred only during the phases of cell growth in the presence of the two carbon sources. The cell-associated glucosyltransferase activities of glucose-, fructose-, mannitol-, and sorbitol-grown cells were relatively low, yet all the cells were capable of adherence to glass in the presence of sucrose. When glucose-containing cultures of S. mutans were supplemented with sucrose, extracellular glucosyltransferase activity first became cell associated and then appeared to become inactivated, presumably due to the accumulation of insoluble glucans.
This work was aimed at studying the effect of different carbon sources in the composition of mineral media on the growth of fungi belonging to the Aspergillus genus and on the fatty acid composition of their lipids. A chemically-defined medium with glucose was shown to be optimal for the growth of 18 Aspergillus strains and for the synthesis of lipids by them. The fatty acid composition of lipids was studied when the fungi grew in media with different carbon sources. The lipids were shown to contain saturated and unsaturated fatty acids with the prevalence of oleic, linoleic and linolenic acids.
An enhanced biological phosphorus removal (EBPR) system was developed in a sequencing batch reactor (SBR) using propionate as the sole carbon source. The microbial community was followed using fluorescence in situ hybridization (FISH) techniques and Candidatus 'Accumulibacter phosphatis' were quantified from the start up of the reactor until steady state. A series of SBR cycle studies was performed when 55% of the SBR biomass was Accumulibacter, a confirmed polyphosphate accumulating organism (PAO) and when Candidatus 'Competibacter phosphatis', a confirmed glycogen-accumulating organism (GAO), was essentially undetectable. These experiments evaluated two different carbon sources (propionate and acetate), and in every case, two different P-release rates were detected. The highest rate took place while there was volatile fatty acid (VFA) in the mixed liquor, and after the VFA was depleted a second P-release rate was observed. This second rate was very similar to the one detected in experiments performed without added VFA.A kinetic and stoichiometric model developed as a modification of Activated Sludge Model 2 (ASM2) including glycogen economy, was fitted to the experimental profiles. The validation and calibration of this model was carried out with the cycle study experiments performed using both VFAs. The effect of pH from 6.5 to 8.0 on anaerobic P-release and VFA-uptake and aerobic P-uptake was also studied using propionate. The optimal overall working pH was around 7.5. This is the first study of the microbial community involved in EBPR developed with propionate as a sole carbon source along with detailed process performance investigations of the propionate-utilizing PAOs.
Biological denitrification from simulated contaminated groundwater using ethanol as an external carbon source was investigated with an entrapped mixed microbial cell (EMMC) system. Ethanol was used as an external carbon source to serve both as the energy source for the nitrate removal reaction and substrate for microorganisms in the system. The hydraulic retention time and chemical oxygen demand (COD)/nitrogen ratio significantly affect nitrate removal performance and, therefore, must be monitored carefully to achieve high rates of nitrate and COD removal. The effect of packing ratios of carriers in the reactor on nitrate removal performance was also investigated. The removal of nitrate and COD in reactors with various carrier packing ratios (12, 22, and 35%) is not significantly different during steady-state operation. However, the start-up periods in these three reactors are significantly different. The highest packing ratio can result in a fast removal of nitrate during the start-up periods. Results of solids retention time (SRT) analysis indicate that a long SRT in the EMMC system contributes to the stable removal of nitrate and to fast start up of the system. The increase of SRT is related to the increase in days of operation, SRT is maintained at a level of more than 200 days.
In our previous study, the first structural gene (GGTI) encoding g-glutamyl transpeptidase was cloned and characterized from the fission yeast Schizosaccharomyces pombe, and its transcription, using the GGTI-lacZ fusion gene, containing the 1,085 bp upstream region from the translational initiation point, was found to be enhanced by sodium nitroprusside and L-buthionine-(S,R)-sulfoximine (BSO). In the present work, regulation of the GGTI gene was further elucidated. Non-fermentable carbon sources, such as acetate and ethanol, markedly enhanced the synthesis of beta-galactosidase from the GGTI-lacZ fusion gene. However, its induction by non-fermentable carbon sources appeared to be independent of the presence of the Pap1 protein. Nitrogen starvation also gave rise to induction of GGTI gene expression in a Pap1-independent manner. The three additional fusion plasmids, carrying 754, 421 and 156 bp regions, were constructed. The sequence responsible for the induction by non-fermentable carbon sources and nitrogen starvation was identified to exist within a -421 bp region of the GGTI gene. Taken together, the S. pombe GGTI gene is regulated by non-fermentable carbon sources and nitrogen starvation.
Growth on a medium containing succinic acid as the sole carbon source produced 1 g (dry weight) of mycelium per liter of medium by 50 days of incubation, whereas 25 g of mycelium was produced in 10 days when glucose was also present in the medium. Primary shunt metabolism took place during growth on succinic acid in spite of the extremely slow growth. Mycelia grown on succinic acid contained a higher percentage of residual mycelium and phosphate, but a lower percentage of mannitol, carbohydrate, lipid, and water-soluble nitrogen, than mycelia grown on a mixture of glucose and succinic acid. Thus, although primary shunt metabolism is favored by rapid growth on a rich, balanced sugar medium, it can also take place during extremely restricted growth in a medium containing succinic acid as the sole carbon source.