RESUMO
BACKGROUND: poly(L-malic acid) (PMA) is a water-soluble polyester with many attractive properties in medicine and food industries, but the high cost of PMA fermentation has restricted its further application for large-scale production. To overcome this problem, PMA production from Jerusalem artichoke tubers was successfully performed. Additionally, a valuable exopolysaccharide, pullulan, was co-produced with PMA by Aureobasidum pullulans HA-4D. RESULTS: The Jerusalem artichoke medium for PMA and pullulan co-production contained only 100 g/L hydrolysate sugar, 30 g/L CaCO3 and 1 g/L NaNO3. Compared with the glucose medium, the Jerusalem artichoke medium resulted in a higher PMA concentration (114.4 g/L) and a lower pullulan concentration (14.3 g/L) in a 5 L bioreactor. Meanwhile, the activity of pyruvate carboxylase and malate dehydrogenas was significantly increased, while the activity of α-phosphoglucose mutase, UDP-glucose pyrophosphorylase and glucosyltransferase was not affected. To assay the economic-feasibility, large-scale production in a 1 t fermentor was performed, yielding 117.5 g/L PMA and 15.2 g/L pullulan. CONCLUSIONS: In this study, an economical co-production system for PMA and pullulan from Jerusalem artichoke was developed. The medium for PMA and pullulan co-production was significantly simplified when Jerusalem artichoke tubers were used. With the simplified medium, PMA production was obviously stimulated, which would be associated with the improved activity of pyruvate carboxylase and malate dehydrogenas.
Assuntos
Ascomicetos/metabolismo , Glucanos/economia , Glucanos/metabolismo , Helianthus/metabolismo , Malatos/economia , Malatos/metabolismo , Polímeros/economia , Polímeros/metabolismo , Ascomicetos/genética , Técnicas de Cultura Celular por Lotes/métodos , Engenharia Genética/economia , Engenharia Genética/métodos , Glucanos/genética , Glucanos/isolamento & purificação , Helianthus/genética , Malatos/isolamento & purificação , Tubérculos/genética , Tubérculos/metabolismo , Polímeros/isolamento & purificaçãoRESUMO
Lignocellulosic biomass and dedicated energy crops such as Jerusalem artichoke are promising alternatives for biobutanol production by solventogenic clostridia. However, fermentable sugars such as fructose or xylose released from the hydrolysis of these feedstocks were subjected to the incomplete utilization by the strains, leading to relatively low butanol production and productivity. When 0.001 g/L ZnSO4·7H2O was supplemented into the medium containing fructose as sole carbon source, 12.8 g/L of butanol was achieved with butanol productivity of 0.089 g/L/h compared to only 4.5 g/L of butanol produced with butanol productivity of 0.028 g/L/h in the control without zinc supplementation. Micronutrient zinc also led to the improved butanol production up to 8.3 g/L derived from 45.2 g/L xylose as sole carbon source with increasing butanol productivity by 31.7%. Moreover, the decreased acids production was observed under the zinc supplementation condition, resulting in the increased butanol yields of 0.202 g/g-fructose and 0.184 g/g-xylose, respectively. Similar improvements were also observed with increasing butanol production by 130.2 % and 8.5 %, butanol productivity by 203.4% and 18.4%, respectively, in acetone-butanol-ethanol fermentations from sugar mixtures of fructose/glucose (4:1) and xylose/glucose (1:2) simulating the hydrolysates of Jerusalem artichoke tubers and corn stover. The results obtained from transcriptional analysis revealed that zinc may have regulatory mechanisms for the sugar transport and metabolism of Clostridium acetobutylicum L7. Therefore, micronutrient zinc supplementation could be an effective way for economic development of butanol production derived from these low-cost agricultural feedstocks.
Assuntos
1-Butanol/metabolismo , Acetona/metabolismo , Etanol/metabolismo , Fermentação , Frutose/metabolismo , Xilose/metabolismo , Zinco/metabolismo , Biomassa , Metabolismo dos Carboidratos/efeitos dos fármacos , Clostridium acetobutylicum/efeitos dos fármacos , Clostridium acetobutylicum/metabolismo , Fermentação/efeitos dos fármacos , Glucose/metabolismo , Helianthus/química , Helianthus/metabolismo , Zea mays/química , Zea mays/metabolismo , Zinco/farmacologiaRESUMO
Copper-contaminated soils were managed with aided phytoextraction in 31 field plots at a former wood preservation site, using a single incorporation of compost (OM) and dolomitic limestone (DL) followed by a crop rotation with tobacco and sunflower. Six amended plots, with increasing total soil Cu, and one unamended plot were selected together with a control uncontaminated plot. The mobility and phytoavailability of Cu, Zn, Cr and As were investigated after 2 and 3 years in soil samples collected in these eight plots. Total Cu, Zn, Cr and As concentrations were determined in the soil pore water (SPW) and available soil Cu and Zn fractions by DGT. The Cu, Zn, Cr and As phytoavailability was characterized by growing dwarf beans on potted soils and determining the biomass of their plant parts and their foliar ionome. Total Cu concentrations in the SPW increased with total soil Cu. Total Cu, Zn, Cr and As concentrations in the SPW decreased in year 3 as compared to year 2, likely due to annual shoot removals by the plants and the lixiviation. Available soil Cu and Zn fractions also declined in year 3. The Cu, Zn, Cr and As phytoavailability, assessed by their concentration and mineral mass in the primary leaves of beans, was reduced in year 3.
Assuntos
Cobre/metabolismo , Helianthus/crescimento & desenvolvimento , Chumbo/metabolismo , Nicotiana/crescimento & desenvolvimento , Poluentes do Solo/metabolismo , Zinco/metabolismo , Biodegradação Ambiental , Biomassa , Cobre/análise , França , Helianthus/metabolismo , Chumbo/análise , Brotos de Planta/crescimento & desenvolvimento , Brotos de Planta/metabolismo , Solo/química , Poluentes do Solo/análise , Nicotiana/metabolismo , Zinco/análiseRESUMO
Bacillus sp. are specific producers of peptidase amongst bacteria and peptidase enzymes and are of significant ones due to their multifarious applications. Advances in industrial biotechnology offer potential opportunities for economic utilization of agro-industrial by-products for many biochemical reactions. Due to their rich organic nature, they can serve as an ideal substrate for the production of different value added products like peptidases. In the present work, an attempt was made to optimize different variables by Taguchi methodology for the production of peptidase using agro-industrial by-products hydrolyzed by a Bacillus cereus strain, resulting in brewer's spent grain (BSG) being the optimal organic substrate. Subsequently, operative variables for the BSG were investigated using Taguchi methodology in order to maximize the enzyme production. Additionally, the main medium components were optimized using a mixture design. Finally, the production of peptidase by B. cereus was investigated; also the possible interaction with other proteolytic microbial strains was evaluated. A notorious synergistic effect was observed when B. cereus was inoculated with Pseudomonas sp. These brought a triple benefit, first, opening the possibility to produce technical enzymes at low cost, second, giving greater value to a food industry by-product, and third, reducing the environmental impact caused by the product removal directly into the environment.
Assuntos
Bacillus cereus/enzimologia , Peptídeo Hidrolases/biossíntese , Pseudomonas/enzimologia , Algoritmos , Meios de Cultura , Fibras na Dieta/metabolismo , Fermentação , Indústria de Processamento de Alimentos , Helianthus/metabolismo , Hidrólise , Peptídeo Hidrolases/isolamento & purificação , Glycine max/metabolismo , Simbiose , ResíduosRESUMO
In Thermus thermophilus HB27 cultures the localisation of lipolytic activity is extracellular, intracellular and membrane bound, with low percentage for the former. Therefore, the extracellular secretion must be increased in order to simplify the downstream process and to reduce the economic cost. This study focuses on the design of an innovative operational strategy to increase extracellular lipolytic enzyme production by T. thermophilus HB27 at bioreactor scale. In order to favour its secretion, the effect of several operational variables was evaluated. Among them, the presence of oils in the culture medium leads to improvements in growth and lipolytic enzyme activity. Sunflower oil is the most efficient inducer showing better results when added after 10h of growth. On the other hand, although surfactants lead to an almost complete inhibition of growth and lipolytic enzyme production, their addition along the culture could affect the location of the enzyme. Thus, by addition of surfactants at the stationary phase, a release of intracellular and membrane enzyme which increases the extracellular enzyme proportion is detected. Based on these results, strategies with successive addition of oil and surfactant in several culture phases in shake flask are developed and verified in a laboratory scale stirred tank bioreactor.
Assuntos
Biotecnologia/métodos , Microbiologia Industrial/métodos , Thermus/enzimologia , Reatores Biológicos , Meios de Cultura/metabolismo , Detergentes/farmacologia , Gases , Helianthus/metabolismo , Concentração de Íons de Hidrogênio , Microbiologia Industrial/economia , Cinética , Lipase/metabolismo , Lipídeos/química , Oxigênio/metabolismo , Óleos de Plantas/metabolismo , Tensoativos/química , Thermus/citologia , Thermus/crescimento & desenvolvimentoRESUMO
2,3-Butanediol is one of the promising bulk chemicals with wide applications. Its fermentative production has attracted great interest due to the high end concentration. However, large-scale production of 2,3-butanediol requires low-cost substrate and efficient fermentation process. In the present study, 2,3-butanediol production by Klebsiella pneumoniae from Jerusalem artichoke tubers was successfully performed, and various technologies, including separate hydrolysis and fermentation (SHF) and simultaneous saccharification and fermentation (SSF), were investigated. The concentration of target products reached 81.59 and 91.63 g/l, respectively after 40 h in batch and fed-batch SSF processes. Comparing with fed-batch SHF, the fed-batch SSF provided 30.3% higher concentration and 83.2% higher productivity of target products. The results showed that Jerusalem artichoke tuber is a favorable substrate for 2,3-butanediol production, and the application of fed-batch SSF for its conversion can result in a more cost-effective process.