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1.
The effect of pH, growth rate, phosphate and iron limitation, carbon monoxide, and carbon source on product formation by Clostridium pasteurianum was determined. Under phosphate limitation, glucose was fermented almost exclusively to acetate and butyrate independently of the pH and growth rate. Iron limitation caused lactate production (38 mol/100 mol) from glucose in batch and continuous culture. At 15% (vol/vol) carbon monoxide in the atmosphere, glucose was fermented to ethanol (24 mol/100 mol), lactate (32 mol/100 mol), and butanol (36 mol/100 mol) in addition to the usual products, acetate (38 mol/100 mol) and butyrate (17 mol/100 mol). During glycerol fermentation, a completely different product pattern was found. In continuous culture under phosphate limitation, acetate and butyrate were produced only in trace amounts, whereas ethanol (30 mol/100 mol), butanol (18 mol/100 mol), and 1,3-propanediol (18 mol/100 mol) were the major products. Under iron limitation, the ratio of these products could be changed in favor of 1,3-propanediol (34 mol/100 mol). In addition, lactate was produced in significant amounts (25 mol/100 mol). The tolerance of C. pasteurianum to glycerol was remarkably high; growth was not inhibited by glycerol concentrations up to 17% (wt/vol). Increasing glycerol concentrations favored the production of 1,3-propanediol. 相似文献
2.
González-Pajuelo M Andrade JC Vasconcelos I 《Journal of industrial microbiology & biotechnology》2004,31(9):442-446
Growth inhibition of Clostridium butyricum VPI 3266 by raw glycerol, obtained from the biodiesel production process, was evaluated. C. butyricum presents the same tolerance to raw and to commercial glycerol, when both are of similar grade, i.e. above 87% (w/v). A 39% increase of growth inhibition was observed in the presence of 100 g l–1 of a lower grade raw glycerol (65% w/v). Furthermore, 1,3-propanediol production from two raw glycerol types (65% w/v and 92% w/v), without any prior purification, was observed in batch and continuous cultures, on a synthetic medium. No significant differences were found in C. butyricum fermentation patterns on raw and commercial glycerol as the sole carbon source. In every case, 1,3-propanediol yield was around 0.60 mol/mol glycerol consumed. 相似文献
3.
Hao J Lin R Zheng Z Sun Y Liu D 《Journal of industrial microbiology & biotechnology》2008,35(12):1615-1624
3-Hydroxypropionaldehyde (3-HPA) is a toxic intermediary metabolite in the biological route of 1,3-propanediol biosynthesis
from glycerol. 3-HPA accumulated in culture medium would arouse an irreversible cessation of the fermentation process. The
role of substrate (glycerol) on 3-HPA accumulation in aerobic fermentation was investigated in this paper. 1,3-Propanediol
oxidoreductase and glycerol dehydratase, two key enzyme catalyzing reactions of 3-HPA formation and consumption, were sensitive
to high concentration of 3-HPA. When the concentration of 3-HPA increased to a higher level in medium (ac 10 mmol/L), the
activity of 1,3-propanediol oxidoreductase in cell decreased correspondingly, which led to decrease of the 3-HPA conversion
rate, then the 3-HPA concentration increasing was accelerated furthermore. 3-HPA accumulation in culture medium was triggered
by this positive feedback mechanism. In the cell exponential growth phase, the reaction catalyzed by 1,3-propanediol oxidoreductase
was the rate limiting step in 1,3-propanediol production. The level of 3-HPA in culture medium could be controlled by the
substrate (glycerol) concentration, and lower level of glycerol could avoid 3-HPA accumulating to a high, lethal concentration.
In fed batch fermentation, under the condition of initial glycerol concentration 30 g/L, and keeping glycerol concentration
lower than 7–8 g/L in cell exponential growth phase, 3-HPA accumulation could not be incurred. Based on this result, a glycerol
feeding strategy was set up in fed batch fermentation. Under the optimized condition, 50.1 g/L of 1,3-propanediol was produced
in 24 h, and 73.1 g/L of final 1,3-propanediol concentration was obtained in 54 h. 相似文献
4.
Dr. E Petitdemange C Dürr S Abbad Andaloussi G Raval 《Journal of industrial microbiology & biotechnology》1995,15(6):498-502
Industrial glycerol obtained through the transesterification process using rapeseed oil did not support growth of several strains ofClostridium butyricum obtained from bacterial culture collections. Ten new strains ofC. butyricum were obtained from mud samples from a river, a stagnant pond, and a dry canal. These new isolates fermented the commercial glycerol and produced 1,3-propanediol as a major fermentation product with concomitant production of acetic and butyric acids. Four of the ten isolates were able to grow on industrial glycerol obtained from rapeseed oil. One strain,C. butyricum E5, was very resistant to high levels of glycerol and 1,3-propanediol. Using fed-batch fermentation, 109 g L–1 of industrial glycerol were converted into 58 g of 1,3-propanediol, 2.2 g of acetate and 6.1 g of butyrate per liter. 相似文献
5.
Biebl H 《Journal of industrial microbiology & biotechnology》2001,27(1):18-26
The fermentation of glycerol by Clostridium pasteurianum was studied with respect to product formation as influenced by the culture conditions. In the majority of batch cultures,
butanol was the main fermentation product, but a varying fraction of glycerol was also converted to 1,3-propanediol, butyric
and acetic acids and ethanol. More than 60 g/l glycerol was utilized, and up to 17 g/l butanol was produced. Fed-batch cultures
did not offer an advantage. When molecular nitrogen was used as a nitrogen source, the fermentation time was prolonged by
a factor of 1.5. Fermentations at constant pH values between 4.5 and 7.5 did not reveal significant differences in product
formation except for an increase in the ethanol content starting at pH 6.5. Chemostat cultures also yielded predominantly
n-butanol, but in some fermentations, the 1,3-propanediol fraction was relatively high. The pH auxostat cultures, which were
operated at a glycerol excess, contained 1,3-propanediol as the main product. As a whole, the fermentations were characterized
by a certain variability in product formation under seemingly equal or slightly varied conditions. It appears that the regulation
of the numerous fermentation pathways occurring in this organism is not very strict. Journal of Industrial Microbiology & Biotechnology (2001) 27, 18–26.
Received 25 September 2000/ Accepted in revised form 07 April 2001 相似文献
6.
Glycerol fermentation of 1,3-propanediol by Clostridium butyricum. Measurement of product inhibition by use of a pH-auxostat 总被引:3,自引:0,他引:3
Hanno Biebl 《Applied microbiology and biotechnology》1991,35(6):701-705
Summary The fermentation of glycerol to 1,3-propanediol, acetate, and butyrate by Clostridium butyricum was studied with respect to growth inhibition by the accumulating products. The clostridia were grown in a pH-auxostat culture at low cell density and product concentration and near maximum growth rate. The products were then added individually to the medium in increasing concentrations and the resulting depression of growth rate was used as a quantitative estimate of product inhibition. Under these conditions growth was totally inhibited at concentrations of 60 g/l for 1,3-propanediol, 27 g/l for acetic acid and 19 g/l for butyric acid at pH 6.5. Appreciable inhibition by glycerol was found only above a concentration of 80 g/l. In a pH-auxostat without added products but with high cell density as well as in batch cultures the product proportions were different. The 1,3-propanediol concentration may approach the value of complete inhibition while the concentrations of acetic and butyric acids remained below these values by at least one order of magnitude. It was therefore concluded that 1,3-propanediol is the first range inhibitor in this fermentation. 相似文献
7.
González-Pajuelo M Meynial-Salles I Mendes F Soucaille P Vasconcelos I 《Applied and environmental microbiology》2006,72(1):96-101
Clostridium acetobutylicum is not able to grow on glycerol as the sole carbon source since it cannot reoxidize the excess of NADH generated by glycerol catabolism. Nevertheless, when the pSPD5 plasmid, carrying the NADH-consuming 1,3-propanediol pathway from C. butyricum VPI 3266, was introduced into C. acetobutylicum DG1, growth on glycerol was achieved, and 1,3-propanediol was produced. In order to compare the physiological behavior of the recombinant C. acetobutylicum DG1(pSPD5) strain with that of the natural 1,3-propanediol producer C. butyricum VPI 3266, both strains were grown in chemostat cultures with glycerol as the sole carbon source. The same "global behavior" was observed for both strains: 1,3-propanediol was the main fermentation product, and the qH2 flux was very low. However, when looking at key intracellular enzyme levels, significant differences were observed. Firstly, the pathway for glycerol oxidation was different: C. butyricum uses a glycerol dehydrogenase and a dihydroxyacetone kinase, while C. acetobutylicum uses a glycerol kinase and a glycerol-3-phosphate dehydrogenase. Secondly, the electron flow is differentially regulated: (i) in C. butyricum VPI 3266, the in vitro hydrogenase activity is 10-fold lower than that in C. acetobutylicum DG1(pSPD5), and (ii) while the ferredoxin-NAD+ reductase activity is high and the NADH-ferredoxin reductase activity is low in C. acetobutylicum DG1(pSPD5), the reverse is observed for C. butyricum VPI 3266. Thirdly, lactate dehydrogenase activity is only detected in the C. acetobutylicum DG1(pSPD5) culture, explaining why this microorganism produces lactate. 相似文献
8.
M. Heyndrickx P. De Vos M. Vancanneyt J. De Ley 《Applied microbiology and biotechnology》1991,34(5):637-642
Summary In batch culture on reiinforced clostridial medium strain-dependent product profiles from glycerol revealed unusual fermentation products such as propionate and n-propanol with Clostridium butyricum LMG 1213t1, and 1,3-propanediol with C. butyricum LMG 1212t2 and C. pasteurianium LMG 3285. Only the latter two strains were able to grow on glycerol in a minimal medium. Nicotinamide adenine dinucleotide (NAD+)-dependent dehydrogenase activities were detected with 1,3-propanediol and n-butanol as substrate (the latter only after a lag period) in cell-free extracts of C. butyricum LMG 1212t2 and with 1,3-propanediol, n-butanol and ethanol in cell-free extracts of C. pasteurianum LMG 3285. The data indicated the existance of a specific 1,3-propanediol dehydrogenase in both organisms. In a chemostat, C. butyricum LMG 1212t2 converted 65% of the glycerol supplied as sole carbon and energy source to 1,3-propanediol without H2 production. Increasing concentration of acetate in the inflow medium resulted in less 1,3-propanediol and more butyrate and H2 production. C. pasteurianum LMG 3285 converted somewhat more than half of the glycerol supplied as sole energy and carbon source to n-butanol with significant concomitant H2 production. This fermentation pattern was hardly affected by acetate as co-substrate.
Offprint requests to: P. De Vos 相似文献
9.
We report a Klebsiella pneumoniae DSM2026 fermentation procedure for the efficient production of a key enzyme of 1,3-propanediol formation: 1,3-propanediol oxidoreductase (E.C. 1.1.1.202). The fermentation process is composed of an aerobic batch phase on glucose and glycerol and an anaerobic phase on glycerol. The role of the aerobic phase is to produce sufficiently high cell mass (12.9–14.6 g/l dry weight) and to activate the aerobic branch of the Klebsiella glycerol pathway, whereas in the anaerobic phase there is a rapid initiation of 1,3-propanediol oxidoreductase formation. A fast change from an aerobic to an anaerobic environment led to a redox imbalance, which resulted in the abrupt activation of the anaerobic branch of glycerol utilization, with the occurrence of a high 1,3-propanediol-oxidoreductase activity. A mathematical model with substrate inhibition showed that the adequate glycerol concentration for enzyme production was 14–16 g/l. The combination of the optimal substrate concentration together with the subsequent use of glucose and glycerol resulted in 90.6 ± 11.6 U enzyme activity referred to 1 l of fermentation broth and 10.3 ± 0.9 U/(1 h) productivity. 相似文献
10.
Rainer Boenigk Susanne Bowien Gerhard Gottschalk 《Applied microbiology and biotechnology》1993,38(4):453-457
The conversion of glycerol to 1,3-propanediol by Citrobacter freundii DSM 30040 was optimized in single- and two-stage continuous cultures. The productivity of 1,3-propanediol formation was highest under glycerol limitation and increased with the dilution rate (D) to a maximum of 3.7 g·l–1·h–1. Glycerol dehydratase seemed to be the rate-limiting step in 1,3-propanediol formation. Conditions for the two-stage fermentation process were as follows: first stage, glycerol limitation (250mM), pH 7.2, D=0.1 h–, 31° C; second stage, additional glycerol, pH 6.6, D=0.05 h–1, 28° C. Under these conditions 875mM glycerol were consumed, the final 1,3-propanediol concentration was 545mM, and the overall productivity 1.38 g·1–1·h–1.
Correspondence to: G. Gottschalk 相似文献
11.
Jensen TO Kvist T Mikkelsen MJ Christensen PV Westermann P 《Journal of industrial microbiology & biotechnology》2012,39(5):709-717
Clostridium pasteurianum can utilize glycerol as the sole carbon source for the production of butanol and 1,3-propanediol. Crude glycerol derived
from biodiesel production has been shown to be toxic to the organism even in low concentrations. By examination of different
pretreatments we found that storage combined with activated stone carbon addition facilitated the utilization of crude glycerol.
A pH-controlled reactor with in situ removal of butanol by gas stripping was used to evaluate the performance. The fermentation
pattern on pretreated crude glycerol was quite similar to that on technical grade glycerol. C. pasteurianum was able to utilize 111 g/l crude glycerol. The average consumption rate was 2.49 g/l/h and maximum consumption rate was
4.08 g/l/h. At the maximal glycerol consumption rate butanol was produced at 1.3 g/l/h. These rates are higher than those
previously reported for fermentations on technical grade glycerol by the same strain. A process including pretreatment and
subsequent fermentation of the crude glycerol could be usable for industrial production of butanol by C. pasteurianum. 相似文献
12.
Summary The influence of glycerol (100 mM) on the growth kinetic and the end-products formation by Lactobacillus reuteri growing on maltose (29 mM) was examined in aerobiosis and anaerobiosis. The presence of glycerol in aerated cultures changed the maltose consumption but not the growth rate nor the fermentation balance. In a nitrogen atmosphere, L. reuteri was not able to grow on maltose alone. In the presence of both, maltose and glycerol, L. reuteri changed its fermentation pattern, producing 58 mM 1,3-propanediol as the major end product. 相似文献
13.
Microbial Conversion of Glycerol to 1,3-Propanediol: Physiological Comparison of a Natural Producer, Clostridium butyricum VPI 3266, and an Engineered Strain, Clostridium acetobutylicum DG1(pSPD5) 总被引:1,自引:0,他引:1
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María Gonzlez-Pajuelo Isabelle Meynial-Salles Filipa Mendes Philippe Soucaille Isabel Vasconcelos 《Applied microbiology》2006,72(1):96-101
Clostridium acetobutylicum is not able to grow on glycerol as the sole carbon source since it cannot reoxidize the excess of NADH generated by glycerol catabolism. Nevertheless, when the pSPD5 plasmid, carrying the NADH-consuming 1,3-propanediol pathway from C. butyricum VPI 3266, was introduced into C. acetobutylicum DG1, growth on glycerol was achieved, and 1,3-propanediol was produced. In order to compare the physiological behavior of the recombinant C. acetobutylicum DG1(pSPD5) strain with that of the natural 1,3-propanediol producer C. butyricum VPI 3266, both strains were grown in chemostat cultures with glycerol as the sole carbon source. The same “global behavior” was observed for both strains: 1,3-propanediol was the main fermentation product, and the qH2 flux was very low. However, when looking at key intracellular enzyme levels, significant differences were observed. Firstly, the pathway for glycerol oxidation was different: C. butyricum uses a glycerol dehydrogenase and a dihydroxyacetone kinase, while C. acetobutylicum uses a glycerol kinase and a glycerol-3-phosphate dehydrogenase. Secondly, the electron flow is differentially regulated: (i) in C. butyricum VPI 3266, the in vitro hydrogenase activity is 10-fold lower than that in C. acetobutylicum DG1(pSPD5), and (ii) while the ferredoxin-NAD+ reductase activity is high and the NADH-ferredoxin reductase activity is low in C. acetobutylicum DG1(pSPD5), the reverse is observed for C. butyricum VPI 3266. Thirdly, lactate dehydrogenase activity is only detected in the C. acetobutylicum DG1(pSPD5) culture, explaining why this microorganism produces lactate. 相似文献
14.
High production of 1,3-propanediol from industrial glycerol by a newly isolated Clostridium butyricum strain 总被引:11,自引:0,他引:11
Papanikolaou S Ruiz-Sanchez P Pariset B Blanchard F Fick M 《Journal of biotechnology》2000,77(2-3):191-208
Batch and continuous cultures of a newly isolated Clostridium butyricum strain were carried out on industrial glycerol, the major by-product of the bio-diesel production process. For both types of cultures, the conversion yield obtained was around 0.55 g of 1,3-propanediol formed per 1 g of glycerol consumed whereas the highest 1,3-propanediol concentration, achieved during the single-stage continuous cultures was 35-48 g l-1. Moreover, the strain presented a strong tolerance at the inhibitory effect of the 1,3-propanediol, even at high concentrations of this substance at the chemostat (e.g. 80 g l-1). 1,3-Propanediol was associated with cell growth whereas acetate and butyrate seemed non growth-associated products. At low and medium dilution rates (until 0.1 h-1), butyrate production was favoured, whereas at higher rates acetate production increased. The maximum 1,3-propanediol volumetric productivity obtained was 5.5 g l-1 h-1. A two-stage continuous fermentation was also carried out. The first stage presented high 1,3-propanediol volumetric productivity, whereas the second stage (with a lower dilution rate) served to further increase the final product concentration. High 1,3-propanediol concentrations were achieved (41-46 g l-1), with a maximum volumetric productivity of 3.4 g l-1 h-1. A cell concentration decrease was reported between the second and the first fermentor. 相似文献
15.
发展可再生能源,尤其是生物能源,具有显著的能量收益和碳减排效益。随着石油等不可再生资源的减少,许多大宗传统石油化工产品正不断被使用可再生原料的生物制造产品替代。生物发酵法生产1,3-丙二醇(1,3-PDO)顺应了这一潮流,具有广阔的发展前景。提高微生物发酵竞争力,优化发酵法生产1,3-PDO水平,势必增加1,3-PDO的生产效益。对肺炎克雷伯氏菌(Klebsiella pneumoniae)发酵法进行1,3-PDO生产的代谢机理、菌株筛选和利用、发酵参数的选择和优化以及发酵工程策略的设计和监测等进行综述,为利用生物柴油副产物甘油生产有重要工业价值的1,3-PDO产品提供参考。 相似文献
16.
F. Barbirato C. Camarasa-Claret J. P. Grivet A. Bories 《Applied microbiology and biotechnology》1995,43(5):786-793
According to their ability to synthesize 1,3-propanediol from glycerol, two species were isolated from the anoxic mud of
a distillery waste-water digestor: Clostridium butyricum and Enterobacter agglomerans. The latter, a facultatively anaerobic gram-negative bacterium, is described for the first time as a microorganism producing
1,3-propanediol from glycerol. The products of glycerol conversion by E. agglomerans were identified using nuclear magnetic resonance. A 20-g/l glycerol solution was fermented mainly to 1,3-propanediol (0.51 mol/mol)
and acetate (0.18 mol/mol). Ethanol, formate, lactate and succinate were formed as by-products. Gas production was very low;
1,3-propanediol production perfectly balanced the oxido-reduction state of the microorganism. Acetate was the predominant
metabolite generating energy for growth. High-glycerol-concentration fermentations (71 g/l and 100 g/l) resulted in an increase
of the 1,3-propanediol yield (0.61 mol/mol) at the expense of lactate and ethanol production. Specific rates of glycerol consumption
and 1, 3-propanediol and acetate production increased whereas the growth rate decreased. The decrease in ATP yield was linearly
correlated with the specific rate of 1,3-propanediol production. Incomplete glycerol consumption (about 40 g/l) was systematically
observed when high glycerol concentrations were used. The unbalanced oxido-reduction state, the low carbon recovery and the
detection of an unknown compound by HPLC observed in these cases indicate the formation of another metabolite, which is possibly
an inhibitory factor.
Received: 17 November 1994 / Accepted: 15 December 1994 相似文献
17.
18.
1,3-丙二醇是一种重要的化工原料,主要作为平台化合物用于合成聚酯,如聚对苯二甲酸丙二醇酯。经基因工程改造的克雷伯氏肺炎杆菌LDH526能以甘油作为唯一碳源合成1,3-丙二醇,最终发酵浓度超过90 g/L。甘油浓度是影响1,3-丙二醇合成的关键因素。为了实现对甘油浓度的精确控制,设计并优化了基于发酵动力学的甘油自动流加策略。通过将底物流加速率与易观察变量p H和发酵时间偶联,实现了发酵过程中甘油流加的自启动和甘油浓度的动态控制。发酵72 h,1,3-丙二醇的浓度可稳定超过95 g/L。自动控制甘油流加的发酵过程具有可重复性、连续性以及人工工作量少的特点,有望从实验室规模扩大到生产规模。 相似文献
19.
Production of 1,3-Propanediol by Clostridium butyricum VPI 3266 in continuous cultures with high yield and productivity 总被引:2,自引:0,他引:2
González-Pajuelo M Andrade JC Vasconcelos I 《Journal of industrial microbiology & biotechnology》2005,32(9):391-396
The effects of dilution rate and substrate feed concentration on continuous glycerol fermentation by Clostridium butyricum VPI 3266, a natural 1,3-propanediol producer, were evaluated in this work. A high and constant 1,3-propanediol yield (around
0.65 mol/mol), close to the theoretical value, was obtained irrespective of substrate feed concentration or dilution rate.
Improvement of 1,3-propanediol volumetric productivity was achieved by increasing the dilution rate, at a fixed feed substrate
concentration of 30, 60 or 70 g l−1. Higher 1,3-propanediol final concentrations and volumetric productivities were also obtained when glycerol feed concentration
was increased from 30 to 60 g l−1, at D=0.05–0.3 h−1, and from 60–70 g l−1, at D=0.05 and 0.1 h−1·30 g l−1 of 1,3-propanediol and the highest reported value of productivity, 10.3 g l−1 h−1, was achieved at D=0.30 h−1 and 60 g l−1 of feed glycerol. A switch to an acetate/butyrate ratio higher than one was observed for 60 g l−1 of feed glycerol and a dilution rate higher than 0.10 h−1; moreover, at D=0.30 h−1 3-hydroxypropionaldehyde accumulation was observed for the first time in the fermentation broth of C. butyricum. 相似文献
20.
Production of 1,3-propanediol from Glycerol by Recombinant <Emphasis Type="Italic">E. coli</Emphasis> Using Incompatible Plasmids System 总被引:2,自引:0,他引:2
1,3-Propanediol (1,3-PD) has numerous applications in polymers, cosmetics, foods, lubricants, and medicines as a bifunctional
organic compound. The genes for the production of 1,3-PD in Klebsiella pneumoniae, dhaB, which encodes glycerol dehydratase, and dhaT, which encodes 1,3-PD oxidoreductase, and gdrAB, which encodes glycerol dehydratase reactivating factor, are naturally under the control of different promoters and are transcribed
in different directions. These genes were coexpressed in E. coli using two incompatible plasmids (pET28a and pET22b) in the presence of selective pressure. The recombinant E. coli coexpressed the glycerol dehydratase, 1,3-propanediol oxidoreductase and reactivating factor for the glycerol dehydratase
at high levels. In a fed-batch fermentation of glycerol and glucose, the recombinant E. coli containing these two incompatible plasmids consumed 14.3 g/l glycerol and produced 8.6 g/l 1,3-propanediol. In the substitution
case of yqhD (encoding alcohol dehydrogenase from E. coli) for dhaT, the final 1,3-propanediol concentration of the recombinant E. coli could reach 13.2 g/l. 相似文献