BTEX catabolism interactions in a toluene-acclimatized biofilter

BTEX substrate interactions for a toluene-acclimatized biofilter consortium were investigated. Benzene, ethylbenzene, o-xylene, m-xylene and p-xylene removal efficiencies were determined at a loading rate of 18.07 g m⁻³h⁻¹ and retention times of 0.5–3.0 min. This was also repeated for toluene in a 1:1 (m/m) ratio mixture (toluene: benzene, ethylbenzene, or xylene ) with each of the other compounds individually to obtain a final total loading of 18.07 g m⁻³h⁻¹. The results obtained were modelled using Michaelis–Menten kinetics and an explicit finite difference scheme to generate v _max and K _m parameters. The v _max/K _m ratio (a measure of the catalytic efficiency, or biodegradation capacity, of the reactor) was used to quantify substrate interactions occurring within the biofilter reactor without the need for free-cell suspended and monoculture experimentation. Toluene was found to enhance the catalytic efficiency of the reactor for p-xylene, while catabolism of all the other compounds was inhibited competitively by the presence of toluene. The toluene-acclimatized biofilter was also able to degrade all of the other BTEX compounds, even in the absence of toluene. The catalytic efficiency of the reactor for compounds other than toluene was in the order: ethylbenzene>benzene>o-xylene>m-xylene>p-xylene. The catalytic efficiency for toluene was reduced by the presence of all other tested BTEX compounds, with the greatest inhibitory effect being caused by the presence of benzene, while o-xylene and p-xylene caused the least inhibitory effect. This work illustrated that substrate interactions can be determined directly from biofilter reactor results without the need for free-cell and monoculture experimentation. Received: 13 April 2000 / Received revision: 20 July 2000 / Accepted: 27 July 2000     

Benzene/toluene/p-xylene degradation. Part II. Effect of substrate interactions and feeding strategies in toluene/benzene and toluene/p-xylene fermentations in a partitioning bioreactor

A two-phase aqueous/organic partitioning bioreactor scheme was used to degrade mixtures of toluene and benzene, and toluene and p-xylene, using simultaneous and sequential feeding strategies. The aqueous phase of the partitioning bioreactor contained Pseudomonas sp. ATCC 55595, an organism able to degrade benzene, toluene and p-xylene simultaneously. An industrial grade of oleyl alcohol served as the organic phase. In each experiment, the organic phase of the bioreactor was loaded with 10.15 g toluene, and either 2.0 g benzene or 2.1 g p-xylene. The resulting aqueous phase concentrations were 50 mg/l, 25 mg/l and 8 mg/l toluene, benzene and p-xylene respectively. The simultaneous fermentation of benzene and toluene consumed these compounds at volumetric rates of 0.024 g l⁻¹ h⁻¹ and 0.067 g l⁻¹ h⁻¹, respectively. The simultaneous fermentation of toluene and p-xylene consumed these xenobiotics at volumetric rates of 0.066 g l⁻¹ h⁻¹ and 0.018 g l⁻¹ h⁻¹, respectively. A sequential feeding strategy was employed in which toluene was added initially, but the benzene or p-xylene aliquot was added only after the cells had consumed half of the initial toluene concentration. This strategy was shown to improve overall degradation rates, and to reduce the stress on the microorganisms. In the sequential fermentation of benzene and toluene, the volumetric degradation rates were 0.056 g l⁻¹ h⁻¹ and 0.079 g l⁻¹ h⁻¹, respectively. In the toluene/p-xylene sequential fermentation, the initial toluene load was consumed before the p-xylene aliquot was consumed. After 12 h in which no p-xylene degradation was observed, a 4.0-g toluene aliquot was added, and p-xylene degradation resumed. Excluding that 12-h period, the microbes consumed toluene and p-xylene at volumetric rates of 0.074 g l⁻¹ h⁻¹ and 0.025 g l⁻¹ h⁻¹, respectively. Oxygen limitation occurred in all fermentations during the rapid growth phase. Received: 16 November 1998 / Received revision: 29 March 1999 / Accepted: 9 April 1999     

Continuous cultures of <Emphasis Type="Italic">Pseudomonas putida</Emphasis> mt-2 overcome catabolic function loss under real case operating conditions

The long-term performance and stability of Pseudomonas putida mt-2 cultures, a toluene-sensitive strain harboring the genes responsible for toluene biodegradation in the archetypal plasmid pWW0, was investigated in a chemostat bioreactor functioning under real case operating conditions. The process was operated at a dilution rate of 0.1 h⁻¹ under toluene loading rates of 259 ± 23 and 801 ± 78 g m⁻³ h⁻¹ (inlet toluene concentrations of 3.5 and 10.9 g m⁻³, respectively). Despite the deleterious effects of toluene and its degradation intermediates, the phenotype of this sensitive P. putida culture rapidly recovered from a 95% Tol⁻ population at day 4 to approx. 100% Tol⁺ cells from day 13 onward, sustaining elimination capacities of 232 ± 10 g m⁻³ h⁻¹ at 3.5 g Tol m⁻³ and 377 ± 13 g m⁻³ h⁻¹ at 10.9 g Tol m⁻³, which were comparable to those achieved by highly tolerant strains such as P. putida DOT T1E and P. putida F1 under identical experimental conditions. Only one type of Tol⁻ variant, harboring a TOL-like plasmid with a 38.5 kb deletion (containing the upper and meta operons for toluene biodegradation), was identified.     

Toluene degradation in the recycle liquid of biotrickling filters for air pollution control

Pollutant degradation in biotrickling filters for waste air treatment is generally thought to occur only in the biofilm. In two experiments with toluene degrading biotrickling filters, we show that suspended microorganisms in the recycle liquid may substantially contribute to the overall pollutant removal. Two days after reactor start up, the overall toluene elimination capacity reached a maximum of 125 g m⁻³ h⁻¹, which was twice that found during prolonged operation. High biodegradation activity in the recycle liquid fully accounted for this short-term peak of pollutant elimination. During steady-state operation, the toluene degradation in the recycle liquid was 21% of the overall elimination capacity, although the amount of suspended biomass was only 1% of the amount of immobilized biomass. The results suggest that biotrickling filter performance may be improved by selecting operating conditions allowing for the development of an actively growing suspended culture. Received: 16 June 1999 / Received revision: 17 November 1999 / Accepted: 15 December 1999     

Toluene degradation in a polyurethane biofilter at high loading

In the present study, toluene elimination in the polyurethane (PU) biofilter during long-term (145 day) operation was characterized, and assessed the effects of changing the inlet loading and space velocity (SV). A very high elimination capacity of 3.7 kg·m⁻³·h⁻¹ was obtained at an inlet loading of 4.0 kg·m^–3·h⁻¹ (inlet toluene concentration of 900 ppmv at a SV of 1,040 h⁻¹). Backwashing with irrigation and compressed air allowed maintenance of a pressure drop of < 80 mm H₂O·m⁻¹-filter at an SV of 830 h⁻¹ and an elimination efficiency of > 90% during the 145 day of operation. In conclusion, the PU biofilter can overcome the problems of clogging caused by excess biomass growth and of low treatment capacities of conventional biofilters.     

Removal of nitrobenzene vapors by a trickling air biofilter

A stable microbial consortium that grew on nitrobenzene (NB) as its sole source of carbon, nitrogen and energy and liberated excess nitrogen as ammonia, was immobilized on a perlite-packed trickling air biofilter. On a sustained basis, the biofilter removed 50 g NB m⁻³ packing h⁻¹ and its operation at pH 8.7 resulted in ammonia stripping, making pH and salinity controls unnecessary. Low maintenance and stable performance during 4 months of continuous operation invite the scale-up of this biofilter for control of NB emissions. Received 12 September 1996/ Accepted in revised form 17 December 1996     

Microbial response and elimination capacity in biofilters subjected to high toluene loadings

Elimination capacity (EC) is frequently used as a performance and design criterion for vapor-phase biofilters without further verification of the microbial quantity and activity. This study was conducted to investigate how biofilters respond to high pollutant loadings and ultimately how this affects the EC of the biofilter. Two identical laboratory-scale biofilters were maintained at an initial toluene loading rate of 46 g m⁻³ h⁻¹ for a period of 24 days. After the initial biofilm development stage, the loading rates were increased to 91 g m⁻³ h⁻¹ and 137 g m⁻³ h⁻¹, respectively. Following a short period of pseudo-steady state, toluene removal efficiencies rapidly declined in both biofilters, with a concurrent decline in both critical and maximum ECs. The decline was mainly due to deterioration in the biodegradation activity of the biofilm and a decline in the toluene-degrading bacterial population within the biofilm phase. The findings imply that high toluene loadings accelerated the deterioration in overall performance due to a rapid accumulation of inactive biomass. As a result, care must be used when relying on EC values for biofilter design and operational purposes, since the values do not appropriately reflect the temporal changes in biodegradation activity and active biomass quantities that can occur in biofilters subjected to high inlet loadings.     

Environmental parameters affecting xylitol production from sugar cane bagasse hemicellulosic hydrolyzate by Candida guilliermondii

The bioconversion of xylose to xylitol by Candida guilliermondii FTI 20037 cultivated in sugar cane bagasse hemicellulosic hydrolyzate was influenced by cell inoculum level, age of inoculum and hydrolyzate concentration. The maximum xylitol productivity (0.75 g L⁻¹ h⁻¹) occurred in tests carried out with hydrolyzate containing 54.5 g L⁻¹ of xylose, using 3.0 g L⁻¹ of a 24-h-old inoculum. Xylitol productivity and cell concentration decreased with hydrolyzate containing 74.2 g L⁻¹ of xylose. Received 02 February 1996/ Accepted in revised form 15 November 1996     

Rhamnose lipids – biosynthesis, microbial production and application potential

Biosurfactants containing rhamnose and β-hydroxydecanoic acid and called rhamnolipids are reviewed with respect to microbial producers, their physiological role, biosynthesis and genetics, and especially their microbial overproduction, physicochemical properties and potential applications. With Pseudomonas species, more than 100 g l⁻¹ rhamnolipids were produced from 160 g l⁻¹ soybean oil at a volumetric productivity of 0.4 g l⁻¹h⁻¹. The individual rhamnolipids are able to lower the surface tension of water from 72 mN m⁻¹ to 25–30 mN m⁻¹ at concentrations of 10–200 mg l⁻¹. After initial testing, rhamnolipids seem to have potential applications in combating marine oil pollution, removing oil from sand and in combating zoosporic phytopathogens. Rhamnolipids are also a source of l-rhamnose, which is already used for the industrial production of high-quality flavor components. Received: 1 July 1998 / Received revision: 11 September 1998 / Accepted: 13 September 1998     

Biofiltration of paint solvent mixtures in two reactor types: overloading by hydrophobic components

Steady-state performance characteristics of a trickle bed reactor (TBR) and a biofilter (BF) in loading experiments with increasing toluene/xylenes inlet concentrations while maintaining a constant loading rate of hydrophilic components (methyl ethyl and methyl isobutyl ketones, acetone, and n-butyl acetate) of 4 g m⁻³ h⁻¹ were evaluated and compared, along with the systems’ dynamic responses. At the same combined substrate loading of 55 g m⁻³ h⁻¹ for both reactors, the TBR achieved more than 1.5 times higher overall removal efficiency (RE_W) than the BF. Increasing the loading rate of aromatics resulted in a gradual decrease of their REs. The degradation rates of acetone and n-butyl acetate were also inhibited at higher loads of aromatics, thus revealing a competition in cell catabolism. A step-drop in loading of aromatics resulted in an immediate increase of RE_W with variations in the TBR, while the new steady-state value in the BF took 6–7 h to achieve. The TBR consistently showed a greater performance than BF in removing toluene and xylenes. Increasing the loading rate of aromatics resulted in a gradual decrease of their REs. The degradation rates of acetone and n-butyl acetate were also lower at higher OL_AROM, revealing a competition in the cell catabolism. The results obtained are consistent with the proposed hypothesis of greater toxic effects under low water content, i.e., in the biofilter, caused by aromatic hydrocarbons in the presence of polar ketones and esters, which may improve the hydrocarbon partitioning into the aqueous phase.     

A bioactive foamed emulsion reactor for the treatment of benzene-contaminated air stream

An adapted bioactive foamed emulsion bioreactor for the treatment of benzene vapor has been developed. In this reactor, bed clogging was resolved by bioactive foam as a substitute of packing bed for interfacial contact of liquid to gaseous phase. The pollutant solubility has been increased using biocompatible organic phase in liquid phase and this reactor can be applied for higher inlet benzene concentration. Experimental results showed a benzene elimination capacity (EC) of 220 g m⁻³ h⁻¹ with removal efficiency (RE) of 85% for benzene inlet concentration of 1–1.2 g m⁻³ at 15 s gas residence time in bioreactor. Assessment of benzene concentration in liquid phase showed that a significant amount of transferred benzene mass has been biodegraded. By optimizing the operational parameters of bioreactor, continuous operation of bioreactor with high EC and RE was demonstrated. With respect to the results, this reactor has the potential to be applied instead of biofilter and biotrickling filters.     

Continuous production of l(+)-lactic acid by Lactobacillus casei in two-stage systems

A two-stage two-stream chemostat system and a two-stage two-stream immobilized upflow packed-bed reactor system were used for the study of lactic acid production by Lactobacillus casei subsp casei. A mixing ratio of D ₁₂/D ₂= 0.5 (D = dilution rate) resulted in optimum production, making it possible to generate continuously a broth with high lactic acid concentration (48 g l⁻¹) and with a lowered overall content of initial yeast extract (5  g l⁻¹), half the concentration supplied in the one-step process. In the two-stage chemostat system, with the first stage at pH 5.5 and 37 °C and a second stage at pH 6.0, a temperature change from 40 °C to 45 °C in the second stage resulted in a 100% substrate consumption at an overall dilution rate of 0.05 h⁻¹. To increase the cell mass in the system, an adhesive strain of L. casei was used to inoculate two packed-bed reactors, which operated with two mixed feedstock streams at the optimal conditions found above. Lactic acid fermentation started after a lag period of cell growth over foam glass particles. No significant amount of free cells, compared with those adhering to the glass foam, was observed during continuous lactic acid production. The extreme values, 57.5 g l⁻¹ for lactic acid concentration and 9.72 g l⁻¹ h⁻¹ for the volumetric productivity, in upflow packed-bed reactors were higher than those obtained for free cells (48 g l⁻¹  and 2.42 g l⁻¹ h⁻¹) respectively and the highest overall l(+)-lactic acid purity (96.8%) was obtained in the two-chemostat system as compared with the immobilized-cell reactors (93%). Received: 4 December 1997 / Received revision: 23 February 1998 / Accepted: 14 March 1998     

Benzene/toluene/p-xylene degradation. Part I. Solvent selection and toluene degradation in a two-phase partitioning bioreactor

A two-phase organic/aqueous reactor configuration was developed for use in the biodegradation of benzene, toluene and p-xylene, and tested with toluene. An immiscible organic phase was systematically selected on the basis of predicted and experimentally determined properties, such as high boiling points, low solubilities in the aqueous phase, good phase stability, biocompatibility, and good predicted partition coefficients for benzene, toluene and p-xylene. An industrial grade of oleyl alcohol was ultimately selected for use in the two-phase partitioning bioreactor. In order to examine the behavior of the system, a single-component fermentation of toluene was conducted with Pseudomonas sp. ATCC 55595. A 0.5-l sample of Adol 85 NF was loaded with 10.4 g toluene, which partitioned into the cell containing 1 l aqueous medium at a concentration of approximately 50 mg/l. In consuming the toluene to completion, the organisms were able to achieve a volumetric degradation rate of 0.115 g l⁻¹ h⁻¹. This system is self-regulating with respect to toluene delivery to the aqueous phase, and requires only feedback control of temperature and pH. Received: 16 November 1998 / Received revision: 28 March 1999 / Accepted: 9 April 1999     

Production of Spirulina sp. in sea water supplemented with anaerobic effluents in outdoor raceways under temperate climatic conditions

The use of untreated sea water supplemented with anaerobic effluents from digested pig waste and sodium bicarbonate was evaluated as a low-cost medium for semi-continuous cultivation of a mixed culture of two Spirulina strains in outdoor raceways under temperate climatic conditions (pond temperature in the range 21–26 °C and light intensity in the range 225–957␣μE m⁻² s⁻¹). The mixed culture had a predominant population (86.6 ± 3.9%) of an atypical Spirulina strain consisting of straight filaments, which appeared spontaneously after the strain with helicoidal trichomes had been subcultured. Morphological studies for the identification of the type and size of trichomes of the two strains (HF and SF) were carried out. The proportions of the two strains were observed to be stable during the monitoring period (30 days). Three different sets of semicontinuous cultures were carried out. Sets 1 and 2 were operated under regime 1 (a single addition of anaerobic effluents at time zero and no pH control) during the same season (June and July) of different years. Set 3 was operated under regime 2 (semi-continuous addition of anaerobic effluents and pH control) during the autumn. A minimum productivity of 3.6 g m⁻² day⁻¹ was obtained at one of the lowest temperatures (22.1 °C) and light intensities (245 μE m⁻² s⁻¹) and a maximum productivity of 10.9 g m⁻² day⁻¹ was observed at the highest temperature (25 °C) and highest average light intensity (618 μE m⁻² s⁻¹) registered for sets 1 and 2. The protein content in the Spirulina biomass harvested from these two sets varied from 17% to 65.6%. In set 3, a maximum productivity of 9.0 g m⁻² day⁻¹ was recorded at an average temperature of 24.4 °C and at an average light intensity of 668 μE m⁻² s⁻¹. The protein content in this set under regime 2 varied within a narrower range than in set 1 and set 2 (from 34.8% to 49.1%), apparently because of a continuous availability of ammonia nitrogen at a level of 30–50 mg l⁻¹. However, in terms of the removal of ammonia nitrogen and chemical oxygen demand, regime 1 was more efficient than regime␣2. Received: 3 September 1996 / Received revision: 19 February 1997 / Accepted: 7 March 1997     

The relative importance of tryptophan-dependent and tryptophan-independent biosynthesis of indole-3-acetic acid in tobacco during vegetative growth

A quantitative study of indole-3-acetic acid (IAA) turnover, and the contribution of tryptophan-dependent and tryptophan-independent IAA-biosynthesis pathways, was carried out using protoplast preparations and shoot apices obtained from wild-type and transgenic, IAA-overproducing tobacco (Nicotiana tabacum L.) plants, during a phase of growth when the level of endogenous IAA was stable. Based on the rate of disappearance of [¹³C₆]IAA, the half-life of the IAA pool was calculated to be 1.1 h in wild-type protoplasts and 0.8 h in protoplasts from the IAA-overproducing line, corresponding to metabolic rates of 59 and 160 pg IAA (μg Chl)⁻¹ h⁻¹, respectively. The rate of conversion of tryptophan to IAA was 15 pg IAA (μg Chl)⁻¹ h⁻¹ in wild-type protoplasts and 101 pg IAA (μg Chl)⁻¹ h⁻¹ in protoplasts from IAA-overproducing plants. In both instances, IAA was metabolised more rapidly than it was synthesised from tryptophan. As the endogenous IAA pools were in a steady state, these findings indicate that IAA biosynthesis via the tryptophan-independent pathway was 44 pg IAA (μg Chl)⁻¹ h⁻¹ and 59 pg IAA (μg Chl)⁻¹ h⁻¹, respectively, in the wild-type and transformed protoplast preparations. In a parallel study with apical shoot tissue, the presumed site of IAA biosynthesis, the rate of tryptophan-dependent IAA biosynthesis exceeded the rate of metabolism of [¹³C₆]IAA despite the steady state of the endogenous IAA pool. The most likely explanation for this anomaly is that, unlike the protoplast system, injection of substrates into the apical tissues did not result in uniform distribution of label, and that at least some of the [²H₅]tryptophan was metabolised in compartments not normally active in IAA biosynthesis. This demonstrates the importance of using experimental systems where labelling of the precursor pool can be strictly controlled. Received: 18 January 2000 / Accepted 24 February 2000     

Effects of nitrogen source and empty bed residence time on the removal of styrene gaseous emissions by biotrickling filtration

The removal of styrene-polluted air emissions by biotrickling filtration was performed to evaluate the influence of using nitrate and urea as a nitrogen source in the nutrient solution supplied to two bioreactors run in parallel under the same operational conditions for 3 months. The use of urea resulted in less biomass content along the packed bed and better performance of the process, with a maximum elimination capacity (EC) of 57.6 g C m⁻³ h⁻¹ (removal efficiency (RE) of 88.3% and empty bed residence time (EBRT) of 60 s), which was around 54% higher than when using nitrate. EBRTs of 60, 30 and 15 s were evaluated with a urea-based nutrient supply. By decreasing the EBRT from 60 to 30 s the styrene concentration that could be treated with REs above 80% was almost the half, from 1,100 to 600 mg C m⁻³, resulting in ECs of 52.8 g C m⁻³ h⁻¹. Working at 15 s was not possible to obtain REs higher than 40% with a maximum EC of 28.5 g C m⁻³ h⁻¹.     

Production of curdlan using sucrose or sugar cane molasses by two-step fed-batch cultivation of Agrobacterium species

Maltose and sucrose were efficient carbon sources for the production of curdlan by a strain of Agrobacterium sp. A two-step, fed-batch operation was designed in which biomass was first produced, followed by curdlan production which was stimulated by nitrogen limitation. There exists an optimal timing for nitrogen limitation for curdlan production in the two-step, fed-batch operation. Maximum curdlan production (60 g L⁻¹) was obtained from sucrose with a productivity of 0.2 g L⁻¹ h⁻¹ when nitrogen was limited at a cell concentration of 16.0 g L⁻¹. It was also noted that the curdlan yield from sucrose was as high as 0.45 g curdlan g⁻¹ sucrose, and the highest specific production rate was 1.0 g curdlan g⁻¹ cells h⁻¹ right after nitrogen limitation. Of particular importance was the use of molasses as a cheap carbon source to produce curdlan in the two-step, fed-batch cultivation. As high as 42 g L⁻¹ of curdlan with a yield of 0.35 g curdlan g⁻¹ total sugar was obtained after 120 h of fed-batch cultivation. Received 20 August 1996/ Accepted in revised form 26 November 1996     

Inhibition of Clostridium butyricum by 1,3-propanediol and diols during glycerol fermentation

1,3-Propanediol inhibition during glycerol fermentation to 1,3-propanediol by Clostridium butyricum CNCM 1211 has been studied. The initial concentration of the 1,3-propanediol affected the growth of the bacterium more than the glycerol fermentation. μ _max was inversely proportional to the initial concentration of 1,3-propanediol (0–65 g l⁻¹). For glycerol at 20 g l⁻¹, the growth and fermentation were completely stopped at an initial 1,3-propanediol concentration of 65 g l⁻¹. However, for an initial 1,3-propanediol concentration of 50 g l⁻¹ and glycerol at 70 g l⁻¹, the final concentration (initial and produced) of 1,3-propanediol reached 83.7 g l⁻¹(1.1 M), with complete consumption of the glycerol. Therefore, during the fermentation, the strain tolerated a 1,3-propanediol concentration higher than the initial inhibitory concentration (65 g l⁻¹). The addition of 1,2-propanediol or 2,3-butanediol (50 g l⁻¹) in the presence of glycerol (50–100 g l⁻¹), showed that 2-diols reduced the μ _max in a similar way to 1,3-propanediol. The measurement of the osmotic pressure of glycerol solutions, diols and diol/glycerol mixtures did not indicate any differences between these compounds. The hypothesis of diol inhibition was discussed. Taking into account the strain tolerance of highly concentrated 1,3-propanediol during fermentation, the fermentation processes for optimising production were considered. Received: 15 November 1999 / Revision received: 1 February 2000 / Accepted: 4 February 2000     

Ethanol adaptation in a thermotolerant yeast strain Kluyveromyces marxianus IMB3

The maximum ethanol concentration produced from glucose in defined media at 45°C by the thermotolerant yeast Kluyveromyces marxianus IMB3 was 44 g L⁻¹. Acclimatisation of the strain through continuous culture at ethanol concentrations up to 80 g L⁻¹, shifted the maximum ethanol concentration at which growth was observed from 40 g L⁻¹ to 70 g L⁻¹. Four isolates were selected from the continuous culture, only one of which produced a significant increase in final ethanol concentration (50 ± 0.4 g L⁻¹), however in subsequent fermentations, following storage on nutrient agar plates, the maximum ethanol concentration was comparable with the original isolate. The maximum specific ethanol production rates (approximately 1.5 g (gh)⁻¹) were also comparable with the original strain except for one isolate (0.7 g (gh)⁻¹). The specific ethanol productivity decreased with ethanol concentration; this decrease correlated linearly (rval 0.92) with cell viability. Due to the transience of induced ethanol tolerance in the strain it was concluded that this was not a valid method for improving final ethanol concentrations or production rates. Received 18 July 1997/ Accepted in revised form 19 February 1998     

Production of polyhydroxybutyrate by Ralstonia eutropha from protein hydrolysates

Polyhydroxybutyrate (PHB) was produced by Ralstonia eutropha DSM 11348 (formerly Alicaligenes eutrophus) in media containing 20–30 g l⁻¹ casein peptone or casamino acids as sole sources of nitrogen. In fermentations using media based on casein peptone, permanent growth up to a cell dry mass of 65 g l⁻¹ was observed. PHB accumulated in cells up to 60%–80% of dry weight. The lowest yields were found in media without any trace elements or with casamino acids added only. The residual cell dry masses were limited to 10–15 g l⁻¹ and did not contain PHB. The highest productivity amounted to 1.2 g PHB l⁻¹ h⁻¹. The mean molecular mass of the biopolymer was determined as 750 kDa. The proportion of polyhydroxyvalerate was less than 0.2% in PHB. The bioprocess was scaled up to a 300-l plant. During a fermentation time of 39 h the cells accumulated PHB to 78% w/w. The productivity was 0.98 g PHB l⁻¹ h¹. Received: 8 July 1998 / Accepted: 26 August 1998