Incorporation of 32P and Growth of Pseudomonad UP-2 on n-Tetracosane

Cultures of the marine pseudomonad UP-2 growing on n-tetracosane contained both free cells and cells bound to the solid hydrocarbon. After separation by filtration through a Whatman no. 1 filter, the numbers of free and bound cells were estimated from the amount of ³²P incorporated into each fraction and the determined value of ³²P incorporation per viable cell in the filtrate (free cells). During the early exponential growth phase, over 80% of the cells were bound to large pieces of n-tetracosane; as the culture approached the stationary phase, the number of bound cells remained constant, whereas free cells continued to accumulate. Pulse-labeling experiments indicated that cells grew both on the surface of the solid and in the aqueous medium. During the growth cycle, a portion of the n-tetracosane which was initially nonfilterable was recovered in the filtrate in a form which was largely cell associated. This cell-associated n-tetracosane was preferentially utilized and could completely account for the observed growth of free cells.     

Sequential Growth of Bacteria on Crude Oil

By modification of the enrichment culture procedure three bacterial strains capable of degrading crude oil in sea water were isolated in pure culture, UP-2, UP-3, and UP-4. Strain UP-2 appears to be highly specialized for growth on crude oil in sea water since it showed strong preference for oil or oil degradation products as substrates for growth, converted 66% of the oil into a form no longer extractable by organic solvents, quantitatively degraded the paraffinic fraction (gas chromatographic analysis), emulsified the oil during exponential growth, and produced 1.6 × 10⁸ cells per mg of oil. After exhaustive growth of UP-2 on crude oil the residual oil supported the growth of UP-3 and UP-4, but not a previously isolated oil-degrading bacterium, RAG-1. Strains UP-2, UP-3, and UP-4 grew on RAG-1-degraded oil (specifically depleted of n-alkanes). The growth of UP-3 and UP-4 on UP-2 and RAG-1-degraded oil resulted in the production of new paraffinic compounds as revealed by gas chromatography. When the four strains were grown either together in a mixed culture or sequentially, there was over 75% oil conversion. By plating on selective media, growth of the individual strains was measured kinetically in the reconstituted mixed culture, revealing competition for common growth substances (UP-2 and RAG-1), enhanced die-off (UP-2), and stabilization (UP-4) during the stationary phase.     

Metabolic regulation in Pseudomonas oxalaticus OX1. Diauxic growth on mixtures of oxalate and formate or acetate

Diauxic growth was observed in batch cultures of Pseudomonas oxalaticus when cells were pregrown on acetate and then transferred to mixtures of acetate and oxalate. In the first phase of growth only acetate was utilized. After the exhaustion of acetate from the medium enzymes involved in the metabolism of oxalate were synthesized during a lag phase of 2 h, followed by a second growth phase on oxalate. When the organism was pregrown on oxalate, oxalate utilization from the mixture with acetate completely ceased after a few hours during which acetate became the preferred substrate. Similar observations were made with formate/oxalate mixtures in which formate was the preferred substrate. Until formate was exhausted, it completely suppressed oxalate metabolism, again resulting in diauxic growth. However, when the organism was pregrown on oxalate and then transferred to mixtures of oxalate and formate, both substrates were utilized simultaneously although the initial rate of oxalate utilization from the mixture was strongly reduced as compared to growth on oxalate alone.Since both preferred substrates cross the cytoplasmic membrane by diffusion, whereas oxalate is accumulated by an inducible, active transport system, the effect of acetate and formate on oxalate transport was studied at different external pH values. At pH 5.5 both substrates completely inhibited oxalate transport. However, at pH 7.5, the pH at which the diauxic growth experiments were performed, formate and acetate did not affect oxalate transport. Growth patterns and enzymes profiles suggest that, at higher pH values, formate and acetate possibly affect oxalate utilization via an effect on the internal pool of oxalyl-CoA, the first product of oxalate metabolism.Abbreviations PMS phenazine methosulphate - RuBPCase ribulosebisphosphate carboxylase - DCPIP 2,6-dichlorophenolindophenol - FDH formate dehydrogenase - p.m.f. protonmotive force     

Nutritional characteristics of a mixotrophic nanoflagellate,Ochromonas sp.

Autotrophic and heterotrophic growth characteristics of a nano-flagellate were investigated. The flagellate,Ochromonas sp., was isolated from the northern Baltic Sea. Autotrophic growth was poor. Axenically pregrown flagellates did not increase significantly in cell number during incubation in different inorganic media. The number of flagellates remained constant 3–5 weeks in cultures kept in the light (100mol m^–2 sec^–1), whereas in the dark, a high mortality rate was found. Uptake of inorganic¹⁴C into an acid-stable fraction indicated thatOchromonas had a functional photosynthetic apparatus. Heterotrophic growth in both liquid medium and medium containing bacteria was rapid. The maximum growth rate corresponded to a generation time of 5.3 hours. Light had no effect on heterotrophic growth. Cells pregrown onEscherichia coli minicells survived without additional bacteria as food when kept in the light, but rapid death occurred in darkness. In conclusion, heterotrophy is the major mechanism to support growth in this species ofOchromonas, but under poor environmental conditions photoautotrophy might be a strategy for survival rather than growth.     

A T7 amber mutant defective in DNA-Binding protein

Summary A T7 amber mutant, UP-2, in the gene for T7 DNA-binding protein was isolated from mutants that could not grow on sup ⁺ ssb-1 bacteria but could grow on glnU ssb-1 and sup ⁺ ssb ⁺bacteria. The mutant phage synthesized a smaller amber polypeptide (28,000 daltons) than T7 wild-type DNA-dinding protein (32,000 daltons). DNA synthesis of the UP-2 mutant in sup ⁺ ssb-1 cells was severely inhibited and the first round of replication was found to be repressed. The abilities for genetic recombination and DNA repair were also low even in permissive hosts compared with those of wild-type phage. Moreover, recombination intermediate T7 DNA molecules were not formed in UP-2 infected nonpermissive cells. The gene that codes for DNA-binding protein is referred to as gene 2.5 since the mutation was mapped between gene 2 and gene 3.     

Nitrification, denitrification, and dechlorination in bleached kraft pulp mill wastewater

This study deals with combining the biologi cal removal of organic halogens with the removal of nitrogen from bleached kraft pulp mill wastewater in fluidized-bed reactors under nitrifying and denitrifying conditions. Untreated and biotreated bleached kraft pulp mill wastewaters had no detrimental effect on nitrification or denitrification. The nitrifying biofilm reactor, pregrown on synthetic inorganic feed with ammonia, removed without a lag phase adsorbable organic halogens [7.2 mg Cl (g biomass volatile solids)⁻¹day⁻¹] from bleached kraft pulp mill wastewater and selected chlorophenols from synthetic wastewater. Electron microscopical examination of the biofilm showed that bacteria, morphologically similar to the nitrifying species Nitrosomonas or Nitrobacter, and Nitrosospira were dominant. The denitrifying fluidized-bed reactor, pregrown on nitrate and methanol, denitrified without a lag phase bleached kraft pulp mill wastewater. Under denitrifying conditions, 35% of the total organic carbon content of untreated bleached kraft pulp mill waste water was removed. The reducing power delivered by untreated bleached kraft pulp mill wastewater for denitrification was 2 mmol electrons/mmol carbon mineralized. Dechlorination under denitrifying conditions was negligible. Received: 21 November 1996 / Received revision: 27 January 1997 / Accepted: 1 February 1997     

一株耐碱高效长链烷烃降解菌C24MT1的筛选及其降解特性

【背景】石油被称为“液体黄金”,人类的工业生产活动在利用其创造巨大社会价值的同时,也对自然环境造成了严重的污染。微生物修复技术是现阶段治理石油类污染有效的手段之一,具有经济、高效、无二次污染等优点。【目的】从受石油污染的土壤中分离高效降解长链烷烃正二十四烷的菌株,探究其降解特性及在微生物修复中的应用前景。【方法】通过形态学及16S rRNA基因测序进行菌株鉴定,采用气相色谱法检测菌株对正二十四烷的降解效果,并结合气相色谱-质谱(gas chromatography-mass spectrometer, GC-MS)分析降解中间产物以推测其潜在代谢途径。【结果】筛选到一株可高效降解正二十四烷的菌株C24MT1,经鉴定为不动杆菌属(Acinetobacter)。该菌株最适降解条件为30 °C、pH 9.0、盐度2 g/L,该条件下生长7 d对9 g/L正二十四烷的降解率高达86.63%;与此同时,菌株在强碱性环境(pH 11.0)中生长良好(OD₆₀₀为0.39)并保持较高烷烃降解率(75.38%),对极端环境具备较强的耐受能力;对降解中间产物进行分析,推断菌株代谢长链烷烃正二十四烷的途径可能包括末端氧化及次末端氧化。【结论】不动杆菌C24MT1具有良好的环境适应能力及烷烃降解能力,在后续微生物菌剂开发和石油类污染土壤的环境修复领域具有巨大的应用前景。本研究可为盐碱地区高浓度石油类污染土壤的修复提供优良菌种,并进一步丰富石油烃类生物降解的菌种资源库。     

Regulation of methylotrophic and heterotrophic metabolism in Arthrobacter P1. Growth on mixtures of methylamine and acetate in batch and continuous cultures

Incubations of Arthrobacter P1 in batch culture in media with mixtures of acetate and methylamine resulted in sequential utilization of the two carbon substrates, but not in diauxic growth. Irrespective of the way cells were pregrown, acetate was the preferred substrate and subsequent studies showed that this is due to the fact that acetate is a strong inhibitor of the methylamine transport system and amine oxidase in Arthrobacter P1. An analysis of enzyme activities in cell-free extracts showed that synthesis of amine oxidase occurred already in the first growth phase with acetate, whereas rapid synthesis of hexulose phosphate synthase was only observed once methylamine utilization started. It is therefore concluded that in Arthrobacter P1 the synthesis of the enzymes specific for methylamine oxidation is not regulated co-ordinately with those involved in formaldehyde fixation, but induced sequentially by methylamine and formaldehyde, respectively.During growth of Arthrobacter P1 on the same mixture in carbon- and energy source-limited continuous cultures both substrates were used simultaneously and completely at dilution rates below the _max on either of these substrates. Addition of methylamine, in concentrations as low as 0.5 mM, to the medium reservoir of an acetate-limited continuous culture (D=0.10 h^-1) already resulted in synthesis of both amine oxidase and hexulose phosphate synthase. In the reverse experiment, addition of acetate to the medium reservoir of a methylamine-limited continuous culture (D=0.10 h^-1), acetate was initially only used as an energy source. Synthesis of the glyoxylate cycle enzymes, however, did occur at acetate concentration in the feed above 7.5–10 mM. This indicates that at acetate concentrations below 10 mM the metabolism of the C₁ substrate methylamine is able to cause a complete repression of the synthesis of the enzymes involved in carbon assimilation from the C₂ substrate acetate.Abbreviations HPS Hexulose phosphate synthase - MS mineral salts - RuMP ribulose monophosphate     

Effects of pulse ultrasonic irradiation on the lag phase of Saccharomyces cerevisiae growth

Aims: This paper presents an analysis of lag phase phenomena in Saccharomyces cerevisiae growth as a function of ultrasonic irradiation. Methods and Results: Pulse irradiation treatments were performed by a 20 kHz ultrasonic transducer with different durations and energies. Data obtained from experiments were then employed to estimate growth parameters by specific transfer function. The significance of the different lag times in response to ultrasonic irradiation was analysed. The results showed that the yeast growth in lag phase responded to the irradiated ultrasonic of 20 min more than the 10 min. The ultrasonic energies between 330 and 360 W s m^?3 could decrease lag time up to 1 h compared to the sample without ultrasonic irradiation. Conversely, the treatments with energies higher than 850 W s m^?3 were able to extend the lag time and decrease the yeast growth. Conclusions: The lag durations of S. cerevisiae were changed significantly by different ultrasonic irradiations, energies and durations. In particular, sufficient irradiation energies reduced the lag time, resulting in accelerated yeast growth. In contrast, high energy could inactivate growth by increasing the lag time. Significance and Impact of the Study: This work provides an alternative technique to either accelerate or inactivate the S. cerevisiae lag phase. The approach can be developed in experiment designed to control the yeast growth by ultrasonic irradiation as assistance in the environments.     

The endogenous metabolism ofFibrobacter succinogenes and its relationship to cellobiose transport,viability and cellulose digestion

Fibrobacter succinogenes S85 digested ballmilled cellulose at a rapid rate (0.10 h^–1), but there was a long lag time if the culture was not transferred daily. WhenF. succinogenes was starved for 100h, a large fraction of the cells (>30%) still bound to cellulose, but the lag time was 150h. The lag time was similar for either cellulose- or cellobiose-grown inocula, and lag times were highly correlated (r ² = 0.91) with a decrease in viable cell number. The number of viable cells declined from 10⁸ to 10⁶ in the first 30h of starvation, and by 72h the viable cell number was less than 10³/ml. Cells growing exponentially on cellobiose had a large pool of polysaccharide, and continuous culture experiments indicated that polysaccharide accumulation was not significantly influenced by the growth rate of the culture (approximately 0.7 mg polysaccharide mg^–1 protein). When the cellobiose was depleted, cellular polysaccharide decreased at first order rate of 0.09 h^–1. The rate of endogenous metabolism was initially 0.08mg polysaccharide mg^–1 protein h^–1, and there was little decline in viability until the rate of endogenous metabolism was less than 0.01 mg polysaccharide mg^–1 protein h^–1. When the rate was less than 0.01 mg polysaccharide mg^–1 protein h^–1, the cells could not maintain a sodium gradient, transport cellobiose or grow. The endogenous metabolic rate needed for cell survival was 20 fold less than the maintenance energy of cells growing in continuous culture (0.01 versus 0.232mg carbohydrate mg^–1 protein h^–1).     

Production of flavour esters by Rhizopus oryzae

Microbial production of different alipathic esters with flavour characteristic has been studied. Lyophilized whole cells of Rhizopus oryzae CBS 112-07 were found to be particularly suitable to catalyse the synthesis of different flavour esters (hexyl acetate, propionate, butyrate, caprylate; geranyl acetate, propionate, butyrate and 2- and 3-methylbutyl acetate, butyrate) in n-heptane. This strain was therefore utilized for the semipreparative production of geranyl butyrate by semicontinous and continous addition of the substrates with satisfactory yields (144 g l^–1 in 264 h and 142 g l^–1 in 48 h respectively).     

Thiosulfate oxidation by obligately heterotrophic bacteria

Thiosulfate was oxidized stoichiometrically to tetrathionate during growth on glucose byKlebsiella aerogenes, Bacillus globigii, B. megaterium, Pseudomonas putida, two strains each ofP. fluorescens andP. aeruginosa, and anAeromonas sp. A gram-negative, rod-shaped soil isolate, Pseudomonad H_w, converted thiosulfate to tetrathionate during growth on acetate. None of the organisms could use thiosulfate as sole energy source. The quantitative recovery of all the thiosulfate supplied to heterotrophic cultures either as tetrathionate alone or as tetrathionate and unused thiosulfate demonstrated that no oxidation to sulfate occurred with any of the strains tested. Two strains ofEscherichia coli did not oxidize thiosulfate. Thiosulfate oxidation in batch culture occurred at different stages of the growth cycle for different organisms:P. putida oxidized thiosulfate during lag and early exponential phase,K. aerogenes oxidized thiosulfate at all stages of growth, andB. megaterium andAeromonas oxidized thiosulfate during late exponential phase. The relative rates of oxidation byP. putida andK. aerogenes were apparently determined by different concentrations of thiosulfate oxidizing enzyme. Thiosulfate oxidation byP. aeruginosa grown in chemostat culture was inducible, since organisms pregrown on thiosulfate-containing media oxidized thiosulfate, but those pregrown on glucose only could not oxidize thiosulfate. Steady state growth yield ofP. aeruginosa in glucose-limited chemostat culture increased about 23% in the presence of 5–22 mM thiosulfate, with complete or partial concomitant oxidation to tetrathionate. The reasons for this stimulation are unclear. The results suggest that heterotrophic oxidation of thiosulfate to tetrathionate is widespread across several genera and may even stimulate bacterial growth in some organisms.     

Ethanol production during D-xylose,L-arabinose,and D-ribose fermentation by Bacteroides polypragmatus

Summary The specific growth rate () during cultivation of Bacteroides polypragmatus in 2.51 batch cultures in 4–5% (w/v) l-arabinose medium was 0.23 h^-1 while that in either d-xylose or d-ribose medium was lower (=0.19 h^-1). Whereas growth on arabinose or xylose occurred after about 6–8 h lag period, growth on ribose commenced after a 30 h lag phase. The maximum substrate utilization rate for arabinose, ribose and xylose in media with an initial substrate concentration of 4–5% (w/v) was 0.77, 0.76, and 0.60 g/l/h respectively. In medium containing a mixture of glucose, arabinose, and xylose, the utilization of all three substrates occurred concurrently. The maximum amount of ethanol produced after 72 h growth in 4–5% (w/v) of arabinose, xylose, and ribose was 9.4, 6.5, and 5.3 g/l, respectively. The matabolic end products (mol/mol substrate) of growth in 4.4% (w/v) xylose medium were 0.73 ethanol, 0.49 acetate, 1.39 CO₂, 1.05 H₂, and 0.09 butyrate.National Research Council of Canada No. 23406     

Metabolic regulation in Pseudomonas oxalaticus OX1

Diauxic growth of Pseudomonas oxalaticus was observed on a mixture of formate and oxalate in batch cultures. In the first phase of growth only formate was used. The capacity to oxidize oxalate appeared during the lag phase of 2–4 h after the exhaustion of formate and was followed by a second phase of growth on oxalate. The rate of autotrophic ¹⁴CO₂ fixation measured in washed cell suspensions decreased markedly in this second growth phase on the addition of oxalate. In mixtures of formate with acetate, glyoxylate or glycollate, simultaneous utilization of both substrates was observed. During growth on acetate plus formate formate-oxidizing capacity remained low. With low acetate concentrations, sufficient formate remained after the exhaustion of acetate to support a second growth phase on formate. This phase followed a 1.5–2 h lag, during which formate-oxidizing capacity increased and the Calvin cycle enzymes were synthesized. In mixtures of formate with glyoxylate or glycollate, the formate-oxidizing capacity was high, formate was oxidized rapidly, and no second growth phase was seen. In these latter mixtures high activities of a membrane-bound, phenazine methosulphate/2,6-dichlorophenolindophenollinked formate dehydrogenase and low activities of the soluble NAD-linked formate dehydrogenase were detected. The synthesis of ribulose-1,5-diphosphate carboxylase was totally repressed during growth on formate plus glycollate and partially repressed on formate plus glyoxylate. The regulation of Calvin cyclus enzymes in Pseudomonas oxalaticus is discussed.     

Anaerobic degradation of toluene in denitrifying Pseudomonas sp.: indication for toluene methylhydroxylation and benzoyl-CoA as central aromatic intermediate

The anaerobic degradation of toluene has been studied with whole cells and by measuring enzyme activities. Cultures of Pseudomonas strain K 172 were grown in mineral medium up to a cell density of 0.5 g of dry cells per liter in fed-batch culture with toluene and nitrate as the sole carbon and energy sources. A molar growth yield of 57 g of cell dry matter formed per mol toluene totally consumed was determined. The mean generation time was 24 h. The redox balance between toluene consumed (oxidation and cell material synthesis) and nitrate consumed (reduction to nitrogen gas and assimilation as NH₃) was 77% of expectation if toluene was completely oxidized; this indicated that the major amount of toluene was mineralized to CO₂. It was tested whether the initial reaction in anaerobic toluene degradation was a carboxylation or a dehydrogenation (anaerobic hydroxylation); the hypothetical carboxylated or hydroxylated intermediates were tested with whole cells applying the method of simultanous adaptation: cells pregrown on toluene degraded benzyl alcohol, benzaldehyde, and benzoic acid without lag, 4-hydroxybenzoate and p-cresol with a 90 min lag phase and phenylacetate after a 200 min lag phase. The cells were not at all adapted to degrade 2-methylbenzoate, 4-methylbenzoate, o-cresol, and m-cresol, nor did these compounds support growth within a few days after inoculation with cells grown on toluene. In extracts of cells anaerobically grown on toluene, benzyl alcohol dehydrogenase, benzaldehyde dehydrogenase, and benzoyl-CoA synthetase (AMP forming) activities were present. The data (1) conclusively show anaerobic growth of a pure culture on tolucne; (2) suggest that toluene is anaerobically degraded via benzoyl-CoA; (3) imply that water functions as the source of the hydroxyl group in a toluene methylhydroxylase reaction.     

Influence of sulphur-containing compounds on the growth of Methanosarcina barkeri in a defined medium

Summary Optimal growth of Methanosarcina barkeri occurred in a defined medium containing methanol when 2.5–4 mM sodium sulphide was added giving a concentration of 0.04–0.06 mM dissolved sulphide (HS^–+S^2–. When the sulphide concentration was too low for optimal growth (e.g., 0.1 mM Na₂S added) the addition of the redox resin Serdoxit acted as a sulphide reservoir and caused a significant stimulation of growth. Furthermore it could be demonstrated that iron sulphide, zinc sulphide or L-methionine could also act as sulphur sources while the addition of sodium sulphate to sulphide-depleted media failed to restore growth. The amino acid L-cysteine (0.85 mM) stimulated growth but could not replace Na₂S.Under optimal cysteine-and sulphide concentrations the generation time of this strain was about 7–9 h during growth on methanol, giving a growth yield of about 0.14 g/g methanol consumed. Different M. barkeri strains were also able to grow under these conditions on acetate (30–50 h doubling time) without a significant lag-phase and with complete substrate consumption even though the inoculum was grown on methanol or H₂–CO₂. When methanol and acetate were present as a mixture in the medium both were used simultaneously.     

Adherence of Pseudomonas aeruginosa to inanimate polymers including biomaterials

Cells of Pseudomonas aeruginosa were adhered to polymethyl methacrylate, polyvinyl acetate, polyvinyl chloride, polyhydroxyethyl methacrylate, mixed-acrylic, silicone, and natural latex materials. Planktonic bacteria and bacteria that adhered to the test materials were compared for their uptake of either L-[3,4,5-³H] leucine or [methyl-³H] thymidine during growth in a minimal medium. Leucine incorporation was reduced and thymidine uptake was negligible in adherent bacteria for up to 8 h following primary attachment by which time cells in the planktonic state showed active uptake of both substrates. These reduced uptake periods correlated with lag phases of growth of adherent cells as determined with a sonication-release plate count procedure and analyses of adenosine triphosphate (ATP). The extent of the lag phase of the adherent populations was dependent on initial densities of adhered cells and the nature of the substratum. Received 02 December 1998/ Accepted in revised form 25 April 1999     

Comparison of spiroplasmas by polyacrylamide gel electrophoresis of cell proteins

Pseudomonas oxalaticus utilized sodium acetate or fructose, in addition to sodium formate known to be assimilated via the reductive pentose phosphate pathway. The generation time during growth on fructose (2 h, 10 min) was considerably shorter than observed for other pseudomonads, which sequentially utilize a phosphoenolpyruvate-dependent phosphotransferase system and 1-phosphofructoninase during growth on fructose. In contrast, extracts prepared from fructose-grownP. oxalaticus contaned enzyme activities indicative of the Entner-Doudoroff pathway, while 1-phosphofructokinase was not found. Our studies indicate thatP. oxalaticus may be useful as a model organism for studies of CO₂ fixation.     

Relationship between substrate inhibition and maintenance energy ofChlamydomonas reinhardtii in heterotrophic culture

The relationship between substrate inhibition and maintenance energy ofChlamydomonas reinhardtii grown heterotrophically on acetate was investigated. At low acetate concentrations (<0.4 g l^–1), where no inhibition of cell growth was observed, the cell growth yield and specific growth rate could be represented by the Pirt model, 1/Y=1/Y _g +m/ with a constant value of maintenance energy coefficient m. However, at high acetate concentrations (>0.4 g l^–1), inhibition of cell growth occurred, in which m became variable and dependent on the acetate concentration. A simple mathematical model was proposed to predict the actual maintenance energy coefficient m in the inhibited cultures and experimentally validated.Author for correspondence     

Hexavalent chromium reduction in <Emphasis Type="Italic">Desulfovibrio vulgaris</Emphasis> Hildenborough causes transitory inhibition of sulfate reduction and cell growth

Desulfovibrio vulgaris Hildenborough is a well-studied sulfate reducer that can reduce heavy metals and radionuclides [e.g., Cr(VI) and U(VI)]. Cultures grown in a defined medium had a lag period of approximately 30 h when exposed to 0.05 mM Cr(VI). Substrate analyses revealed that although Cr(VI) was reduced within the first 5 h, growth was not observed for an additional 20 h. The growth lag could be explained by a decline in cell viability; however, during this time small amounts of lactate were still utilized without sulfate reduction or acetate formation. Approximately 40 h after Cr exposure (0.05 mM), sulfate reduction occurred concurrently with the accumulation of acetate. Similar amounts of hydrogen were produced by Cr-exposed cells compared to control cells, and lactate was not converted to glycogen during non-growth conditions. D. vulgaris cells treated with a reducing agent and then exposed to Cr(VI) still experienced a growth lag, but the addition of ascorbate at the time of Cr(VI) addition prevented the lag period. In addition, cells grown on pyruvate displayed more tolerance to Cr(VI) compared to lactate-grown cells. These results indicated that D. vulgaris utilized lactate during Cr(VI) exposure without the reduction of sulfate or production of acetate, and that ascorbate and pyruvate could protect D. vulgaris cells from Cr(VI)/Cr(III) toxicity. J.D. Wall and M.W. Fields are both affiliated to the Virtual Institute of Microbial Stress and Survival (). M.E. Clark and S.B. Thieman contributed equally to this work.