Succinic acid has recently emerged as an important chemical (commodity) because it can be used for the manufacturing of synthetic resins and biodegradable polymers and as an intermediate for chemical synthesis. Till...Succinic acid has recently emerged as an important chemical (commodity) because it can be used for the manufacturing of synthetic resins and biodegradable polymers and as an intermediate for chemical synthesis. Till date, succinic acid is mainly produced by chemical processes, however, due to the environmental concerns and the concepts of sustainability, researches are directed towards the production of succinic acid by microbial fermentation. The fact that carbon dioxide (CO2) is needed by the microorganisms for succinic acid production is another interesting feature. The fermentation was carried out with Actinobacillus succinogenes using a two-level fractional factorial design 2sl. The variables analyzed and their levels were: concentration of glucose, yeast extract, temperature, pH and agitation. The results show that the variables that more influenced on succinic acid production were pH, temperature and yeast extract.展开更多
Succinic acid is considered as an important platform chemical. Succinic acid fermentation with Actinobacillus succinogenes strain BE-1 was optimized by central composite design (CCD) using a response surface methodo...Succinic acid is considered as an important platform chemical. Succinic acid fermentation with Actinobacillus succinogenes strain BE-1 was optimized by central composite design (CCD) using a response surface methodology (RSM). The optimized production of succinic acid was predicted and the interactive effects between glucose, yeast extract, and magnesium carbonate were investigated. As a result, a model for predicting the concentration of succinic acid production was developed. The accuracy of the model was confirmed by the analysis of variance (ANOVA), and the validity was further proved by verification experiments showing that percentage errors between actual and predicted values varied from 3.02% to 6.38%. In addition, it was observed that the interactive effect between yeast extract and magnesium carbonate was statistically significant. In conclusion, RSM is an effective and useful method for optimizing the medium components and investigating the interactive effects, and can provide valuable information for succinic acid scale-up fermentation using A. succinogenes strain BE-I.展开更多
The glutamate decarboxylase(Gad)system is an important amino acid-dependent acid resistance system commonly found in microorganisms.Actinobacillus succinogenes is one of the best natural producers of succinic acid(SA)...The glutamate decarboxylase(Gad)system is an important amino acid-dependent acid resistance system commonly found in microorganisms.Actinobacillus succinogenes is one of the best natural producers of succinic acid(SA)but lacks glutamate decarboxylase.This study assessed the efects of Gad system introduction into A.succinogenes.The recombinant strains gadB-SW and gadBC-SW were constructed by heterologous expression of gadB alone,or gadB together with gadC,respectively.After 1.0 and 1.5 h of acid stress at pH 4.6,cell survival of gadBC-SW was greater than gadB-SW.The growth of gadB-SW and gadBC-SW was both afected by the expression of heterologous proteins and byγ-aminobutyric acid,with gadBC-SW growth reduced at a neutral pH.SA production in acidic conditions was evaluated by a shake fask and by 3-L bioreactor fermentation.The results showed gadBC-SW to increase SA production by 8.4%in shake fask compared to the parent strain,SW.For a 3-L bioreactor batch fermentation under acidic environment,the highest conversion rate of sugar to SA was observed for gadBC-SW,reaching 96%.However,SA concentration by gadBC-SW was only 47 g/L and 31 g/L at pH 6.5 and pH 6.0,respectively.In summary,the introduction of heterologous gadB and gadC into A.succinogenes not only improved acid tolerance but also infuenced the synthesis of SA and added a metabolic burden.展开更多
In this work, the semi-continuous succinic acid fermentation by Actinobacillus succinogenes CGMCC1593 from cane molasses with a novel two-stage and two-stream system was studied, in order to increase the succinic acid...In this work, the semi-continuous succinic acid fermentation by Actinobacillus succinogenes CGMCC1593 from cane molasses with a novel two-stage and two-stream system was studied, in order to increase the succinic acid productivity.The fermentation conditions of the first stage, such as initial sugar concentration, medium addition volume, and time interval of each loading were optimized.Under the optimized condition the maximum succinic acid productivity (2.38 g·L-1·h-1) was increased by 111% and 114% compared with those obtained in batch and fed batch fermentation, respectively.In addition, the maximum succinic acid concentration (46.0 g·L-1)increased by 12.9% compared with that in batch fermentation, and reached a compatible level of that obtained in fed batch fermentation (36 h, 48.8 g·L-1).This process was operated stably for 39 circles without any decreases in both succinic acid concentration and productivity.During the 39 operation circles, the fermentation time of each batch (stage 1 and stage 2), the average succinic acid concentration, succinic acid productivity, succinic acid yield, and sugar conversion rate were 21—24 h, 43.5 g·L-1, 2.07 g·L-1·h-1, 0.79 g·g-1, and 96.4%, respectively.展开更多
Succinic acid is valued as a potential starting point for the production of chemicals of the C4 family or in the prepara-tion of biodegradable polymers. For sustainable development in this era of petroleum shortage, p...Succinic acid is valued as a potential starting point for the production of chemicals of the C4 family or in the prepara-tion of biodegradable polymers. For sustainable development in this era of petroleum shortage, production of succinic acid by microbial fermentation of renewable feedstock has attracted great interest. In this study, pretreatment with sulfuric acid and biotin supplementation were used to enhance succinic acid production by Actinobacillus succinogenes 130Z from sake lees, a byproduct of Japanese rice wine. Pretreatment with sulfuric acid resulted in little change of glucose, total nitrogen and succinic acid content in the sake lees hydrolysate but had a positive effect on succinic acid fermentation, which caused a 25.0% increase in succinic acid yield in batch fermentation. Biotin supplementation was used to further enhance the fermentability of sake lees hydrolysate. As a result, a 30 h batch fermentation of 0.5% sulfuric acid pretreated sake lees hydrolysate with 0.2 mg/L biotin gave a succinic acid yield of 0.59 g/g from 61.6 g/L of glucose, with a productivity of 1.21 g/(L?h). A 22.9% increase in succinic acid yield and a 101.7% increase in succinic acid productivity were obtained compared with untreated sake lees hydrolysate.展开更多
Utilizing CO_(2)for bio-succinic acid production is an attractive approach to achieve carbon capture and recycling(CCR)with simultaneous production of a useful platform chemical.Actinobacillus succinogenes and Basfia ...Utilizing CO_(2)for bio-succinic acid production is an attractive approach to achieve carbon capture and recycling(CCR)with simultaneous production of a useful platform chemical.Actinobacillus succinogenes and Basfia succiniciproducens were selected and investigated as microbial catalysts.Firstly,the type and concentration of inorganic carbon concentration and glucose concentration were evaluated.6 g C/L MgCO_(3)and 24 g C/L glucose were found to be the optimal basic operational conditions,with succinic acid production and carbon yield of over 30 g/L and over 40%,respectively.Then,for maximum gaseous CO_(2)fixation,carbonate was replaced with CO_(2)at different ratios.The“less carbonate more CO_(2)”condition of the inorganic carbon source was set as carbonate:CO_(2)=1:9(based on the mass of carbon).This condition presented the highest availability of CO_(2)by well-balanced chemical reaction equilibrium and phase equilibrium,showing the best performance with regarding CO_(2)fixation(about 15 mg C/(L·hr)),with suppressed lactic acid accumulation.According to key enzymes analysis,the ratio of phosphoenolpyruvate carboxykinase to lactic dehydrogenase was enhanced at high ratios of gaseous CO_(2),which could promote glucose conversion through the succinic acid path.To further increase gaseous CO_(2)fixation and succinic acid production and selectivity,stepwise CO_(2)addition was evaluated.50%-65%increase in inorganic carbon utilization was obtained coupled with 20%-30%increase in succinic acid selectivity.This was due to the promotion of the succinic acid branch of the glucose metabolism,while suppressing the pyruvate branch,along with the inhibition on the conversion from glucose to lactic acid.展开更多
文摘Succinic acid has recently emerged as an important chemical (commodity) because it can be used for the manufacturing of synthetic resins and biodegradable polymers and as an intermediate for chemical synthesis. Till date, succinic acid is mainly produced by chemical processes, however, due to the environmental concerns and the concepts of sustainability, researches are directed towards the production of succinic acid by microbial fermentation. The fact that carbon dioxide (CO2) is needed by the microorganisms for succinic acid production is another interesting feature. The fermentation was carried out with Actinobacillus succinogenes using a two-level fractional factorial design 2sl. The variables analyzed and their levels were: concentration of glucose, yeast extract, temperature, pH and agitation. The results show that the variables that more influenced on succinic acid production were pH, temperature and yeast extract.
基金supported by the National Major Special Project on New Varieties Cultivation for Transgenic Organisms(No.2009ZX08009-130B)the National Natural Science Foundation of China(No.50874112)the National High-Tech R&D Program(863)of China(No.2009AA06Z320)
文摘Succinic acid is considered as an important platform chemical. Succinic acid fermentation with Actinobacillus succinogenes strain BE-1 was optimized by central composite design (CCD) using a response surface methodology (RSM). The optimized production of succinic acid was predicted and the interactive effects between glucose, yeast extract, and magnesium carbonate were investigated. As a result, a model for predicting the concentration of succinic acid production was developed. The accuracy of the model was confirmed by the analysis of variance (ANOVA), and the validity was further proved by verification experiments showing that percentage errors between actual and predicted values varied from 3.02% to 6.38%. In addition, it was observed that the interactive effect between yeast extract and magnesium carbonate was statistically significant. In conclusion, RSM is an effective and useful method for optimizing the medium components and investigating the interactive effects, and can provide valuable information for succinic acid scale-up fermentation using A. succinogenes strain BE-I.
基金The authors are grateful for the fnancial support from the National First-class Discipline Program of Light Industry Technology and Engineering(Grant No.LITE2018-04)the Topnotch Academic Programs Project of Jiangsu Higher Education Institutions(TAPP).
文摘The glutamate decarboxylase(Gad)system is an important amino acid-dependent acid resistance system commonly found in microorganisms.Actinobacillus succinogenes is one of the best natural producers of succinic acid(SA)but lacks glutamate decarboxylase.This study assessed the efects of Gad system introduction into A.succinogenes.The recombinant strains gadB-SW and gadBC-SW were constructed by heterologous expression of gadB alone,or gadB together with gadC,respectively.After 1.0 and 1.5 h of acid stress at pH 4.6,cell survival of gadBC-SW was greater than gadB-SW.The growth of gadB-SW and gadBC-SW was both afected by the expression of heterologous proteins and byγ-aminobutyric acid,with gadBC-SW growth reduced at a neutral pH.SA production in acidic conditions was evaluated by a shake fask and by 3-L bioreactor fermentation.The results showed gadBC-SW to increase SA production by 8.4%in shake fask compared to the parent strain,SW.For a 3-L bioreactor batch fermentation under acidic environment,the highest conversion rate of sugar to SA was observed for gadBC-SW,reaching 96%.However,SA concentration by gadBC-SW was only 47 g/L and 31 g/L at pH 6.5 and pH 6.0,respectively.In summary,the introduction of heterologous gadB and gadC into A.succinogenes not only improved acid tolerance but also infuenced the synthesis of SA and added a metabolic burden.
文摘In this work, the semi-continuous succinic acid fermentation by Actinobacillus succinogenes CGMCC1593 from cane molasses with a novel two-stage and two-stream system was studied, in order to increase the succinic acid productivity.The fermentation conditions of the first stage, such as initial sugar concentration, medium addition volume, and time interval of each loading were optimized.Under the optimized condition the maximum succinic acid productivity (2.38 g·L-1·h-1) was increased by 111% and 114% compared with those obtained in batch and fed batch fermentation, respectively.In addition, the maximum succinic acid concentration (46.0 g·L-1)increased by 12.9% compared with that in batch fermentation, and reached a compatible level of that obtained in fed batch fermentation (36 h, 48.8 g·L-1).This process was operated stably for 39 circles without any decreases in both succinic acid concentration and productivity.During the 39 operation circles, the fermentation time of each batch (stage 1 and stage 2), the average succinic acid concentration, succinic acid productivity, succinic acid yield, and sugar conversion rate were 21—24 h, 43.5 g·L-1, 2.07 g·L-1·h-1, 0.79 g·g-1, and 96.4%, respectively.
文摘Succinic acid is valued as a potential starting point for the production of chemicals of the C4 family or in the prepara-tion of biodegradable polymers. For sustainable development in this era of petroleum shortage, production of succinic acid by microbial fermentation of renewable feedstock has attracted great interest. In this study, pretreatment with sulfuric acid and biotin supplementation were used to enhance succinic acid production by Actinobacillus succinogenes 130Z from sake lees, a byproduct of Japanese rice wine. Pretreatment with sulfuric acid resulted in little change of glucose, total nitrogen and succinic acid content in the sake lees hydrolysate but had a positive effect on succinic acid fermentation, which caused a 25.0% increase in succinic acid yield in batch fermentation. Biotin supplementation was used to further enhance the fermentability of sake lees hydrolysate. As a result, a 30 h batch fermentation of 0.5% sulfuric acid pretreated sake lees hydrolysate with 0.2 mg/L biotin gave a succinic acid yield of 0.59 g/g from 61.6 g/L of glucose, with a productivity of 1.21 g/(L?h). A 22.9% increase in succinic acid yield and a 101.7% increase in succinic acid productivity were obtained compared with untreated sake lees hydrolysate.
基金supported by the Beijing Nova Program(No.Z201100006820022)the National Natural Science Foundation of China(Nos.52100023,U20B2022)+2 种基金the China Postdoctoral Science Foundation(No.2020M680314)the Key Science and Technology Development Program of Xinjiang Corps(No.2021DB006)the EU ACT Coo CE ERA-NET program cofunded with EUDP-2021-Ⅱ(No.64021-2006)。
文摘Utilizing CO_(2)for bio-succinic acid production is an attractive approach to achieve carbon capture and recycling(CCR)with simultaneous production of a useful platform chemical.Actinobacillus succinogenes and Basfia succiniciproducens were selected and investigated as microbial catalysts.Firstly,the type and concentration of inorganic carbon concentration and glucose concentration were evaluated.6 g C/L MgCO_(3)and 24 g C/L glucose were found to be the optimal basic operational conditions,with succinic acid production and carbon yield of over 30 g/L and over 40%,respectively.Then,for maximum gaseous CO_(2)fixation,carbonate was replaced with CO_(2)at different ratios.The“less carbonate more CO_(2)”condition of the inorganic carbon source was set as carbonate:CO_(2)=1:9(based on the mass of carbon).This condition presented the highest availability of CO_(2)by well-balanced chemical reaction equilibrium and phase equilibrium,showing the best performance with regarding CO_(2)fixation(about 15 mg C/(L·hr)),with suppressed lactic acid accumulation.According to key enzymes analysis,the ratio of phosphoenolpyruvate carboxykinase to lactic dehydrogenase was enhanced at high ratios of gaseous CO_(2),which could promote glucose conversion through the succinic acid path.To further increase gaseous CO_(2)fixation and succinic acid production and selectivity,stepwise CO_(2)addition was evaluated.50%-65%increase in inorganic carbon utilization was obtained coupled with 20%-30%increase in succinic acid selectivity.This was due to the promotion of the succinic acid branch of the glucose metabolism,while suppressing the pyruvate branch,along with the inhibition on the conversion from glucose to lactic acid.