[Objective] This study was to realize high-density culture of Chlorella sp. as well as the culture with high lipid yield. [Method] Through analyzing the growth curve of Chlorella sp. cells, dextrose consumption curve,...[Objective] This study was to realize high-density culture of Chlorella sp. as well as the culture with high lipid yield. [Method] Through analyzing the growth curve of Chlorella sp. cells, dextrose consumption curve, change curves of pH and dissolved oxygen, a fed-batch culture was conducted, followed by a nitrogen-deficient culture aiming at accumulating the lipids in Chlorella sp. cells when a high density of Chlorella sp. cells was obtained. [Result] After four batches of feeding were pro- vided, the biomass of Chlorella sp. reached up to 65.25 g/L, and the lipid content increased from 42.75% to 63.82% in Chlorella sp. cells, with the yield of 43.37 g/L in the following 12 hours of nitrogen-deficient culture. [Conclusion] Reasonable fed- batch can significantly improve the biomass of Chlorella sp., and the nitrogen-defi- cient culture further raises the lipid yield of Chlorella sp.展开更多
Recombinant E. coli BL 21 was cultivated in high cell density to produce human-like collagen. The effects of the feeding of nitrogen source, controlled by an auto on/off-feeding mode with two different cycles of 0.5mi...Recombinant E. coli BL 21 was cultivated in high cell density to produce human-like collagen. The effects of the feeding of nitrogen source, controlled by an auto on/off-feeding mode with two different cycles of 0.5min and 4min intervals, oxygen-enrichment methods and inducement strength on the cell yield and human-like collagen production were investigated. The studies showed that nitrogen source feeding in fast cycle could result in higher human-like collagen production than that in slow cycle; and the feedback regulation of glucose, increase of the pressure of fermentation bioreactor, and supply of oxygen-enriched air could all increase cell yield and human-like collagen production. The effects of inducement strength on protein expression were found important. When OD600 reached 90-100, the cultivation temperature was increased to 42℃ to begin induction for 2-3 h, and then shifted to 39℃ for 5-6h induction, the cell density and human-like collagen production could reach 96g·L-1 [DCW (dry cell mass)] and 19.8% (g·g-1 DCW) respectively.展开更多
The production of butyric acid by Clostridium butyricum ZJUCB at various pH values was investigated. In order to study the effect of pH on cell growth, butyric acid biosynthesis and reducing sugar consumption, differe...The production of butyric acid by Clostridium butyricum ZJUCB at various pH values was investigated. In order to study the effect of pH on cell growth, butyric acid biosynthesis and reducing sugar consumption, different cultivation pH values ranging from 6.0 to 7.5 were evaluated in 5-L bioreactor. In controlled pH batch fermentation, the optimum pH for cell growth and butyric acid production was 6.5 with a cell yield of 3.65 g/L and butyric acid yield of 12.25 g/L. Based on these results, this study then compared batch and fed-batch fermentation of butyric acid production at pH 6.5. Maximum value (16.74 g/L) of butyric acid concentration was obtained in fed-batch fermentation compared to 12.25 g/L in batch fermentation. It was concluded that culti- vation under fed-batch fermentation mode could enhance butyric acid production significantly (P<0.01) by C. butyricum ZJUCB.展开更多
An iterative optimization strategy for fed-batch fermentation process is presented by combining a run-to-run optimization with swarm energy conservation particle swarm optimization (SEC-PSO). SEC-PSO, which is designe...An iterative optimization strategy for fed-batch fermentation process is presented by combining a run-to-run optimization with swarm energy conservation particle swarm optimization (SEC-PSO). SEC-PSO, which is designed with the concept of energy conservation, can solve the problem of premature convergence frequently appeared in standard PSO algorithm by partitioning its population into several sub-swarms according to the energy of the swarm and is used in the optimization strategy for parameter identification and operation condition optimization. The run-to-run optimization exploits the repetitive nature of fed-batch processes in order to deal with the optimal problems of fed-batch fermentation process with inaccurate process model and unsteady process state. The kinetic model parameters, used in the operation condition optimization of the next run, are adjusted by calculating time-series data obtained from real fed-batch process in the run-to-run optimization. The simulation results show that the strategy can adjust its kinetic model dynamically and overcome the instability of fed-batch process effectively. Run-to-run strategy with SEC-PSO provides an effective method for optimization of fed-batch fermentation process.展开更多
L-(+)-Lactic acid production from corncob hydrolysate as a cheap carbohydrate source by fed-batch fermentation of Rhizopus oryzaeHZS6 was studied. After 96 h of fermentation in a 5 L fermentor, the final concentration...L-(+)-Lactic acid production from corncob hydrolysate as a cheap carbohydrate source by fed-batch fermentation of Rhizopus oryzaeHZS6 was studied. After 96 h of fermentation in a 5 L fermentor, the final concentration of ammonium L-(+)-lactate, average productivity(based on initial xylose concentration) and maximum dry cell weight were 132.4 g/L, 1.38 g/(L·h), and 8.9 g/L respectively. The optical purity of L-(+)-lactate was 98.8%.展开更多
5-Aminolevulinic acid （ALA） is a common precursor for tetrapyrrole compounds in all kinds of organ isms and has wide applications in agriculture and medicines. In this study, a new strategy, i.e. short-term dissolve...5-Aminolevulinic acid （ALA） is a common precursor for tetrapyrrole compounds in all kinds of organ isms and has wide applications in agriculture and medicines. In this study, a new strategy, i.e. short-term dissolved oxygen （DO） shock during aerobic fermentation, was introduced to produce 5-aminolevulinic acid with a recombi-nant E. coli. Effects of duration time of DO shock operation on plasmid concentration, intracellular ALA synthase （ALAS） activity and ALA production were investigated in Erlenmeyer shake flasks. The results indicated that both ALAS activity and ALA yield were enhanced in an anaerobic operation of 45 rain in the early exponential phase during fermentation, while they decreased when the anaerobic operation time was further increased to 60 rain. The DO shock protocol was confirmed with the fed-batch fermentation in a 15 L fermenter and the ALA production achieved 9.4 g.L-1 （72 mmol.L-1）, which is the highest yield in the fermentation broth reported up to now.展开更多
In this paper,a novel fuzzy neural network model,in which an adjustable fuzzy sub-space was designed by uniform design,has been established and used in fed-batch yeast fermentationas an example.A brand-new optimizatio...In this paper,a novel fuzzy neural network model,in which an adjustable fuzzy sub-space was designed by uniform design,has been established and used in fed-batch yeast fermentationas an example.A brand-new optimization sub-network with special structure has been built andgenetic algorithm,guaranteeing the optimization in overall space,is introduced for the feed rateoptimization.On the basis of the model network,the optimal substrate concentration and theoptimal amount of fed-batch at different periods have been studied,aided with the optimizationnetwork and the genetic algorithm separately.The above results can be used as a basis for theestablishment of a fuzzy neural network controller.展开更多
In this study, Saccharomyces cerevisiae (baker's yeast) was produced in a fed-batch bioreactor at the optimal dissolved oxygen concentration (DOC) and growth medium temperature. However, it is very difficult to co...In this study, Saccharomyces cerevisiae (baker's yeast) was produced in a fed-batch bioreactor at the optimal dissolved oxygen concentration (DOC) and growth medium temperature. However, it is very difficult to control the DOC using conventional controllers because of the poorly understood and constantly changing dynamics of the bioprocess. A generalized predictive controller (GPC) based on a nonlinear autoregressive integrated moving average exogenous (NARIMAX) model is presented to stabilize the DOC by manipulation of air flow rate. The NARIMAX model is built by an improved recursive least-squares support vector machine, which is trained by an in-place computation scheme and avoids the computation of the inverse of a large matrix and memory reallocation. The proposed nonlinear GPC algorithm requires little preliminary knowledge of the fermentation process, and directly obtains the nonlinear model in matrix form by using iterative multiple modeling instead of linearization at each sampling period. By application of an on-line bioreactor control, experimental results demonstrate the robustness, effectiveness and advantages of the new controller.展开更多
An alkalophilc and thermophilic Bacillus sp. BHA that produced a thermostable alkaline protease was isolated from decaying protein substrates. The isolate was found to grow in pH range 7 - 11 with an optimum pH 9.0 an...An alkalophilc and thermophilic Bacillus sp. BHA that produced a thermostable alkaline protease was isolated from decaying protein substrates. The isolate was found to grow in pH range 7 - 11 with an optimum pH 9.0 and temperature up to 55℃. The activity of alkaline protease of Bacillus sp. BHA (68.98 APU/ml) was found higher than the standard strains of Bacillus amyloliquefaciens MTCC 610 (8.98 APU/ml) and Bacillus subtilis MTCC 8349 (12.14 APU/ml, used in this study, and was comparable (68.98 APU/ml, equivalent to 30.38 APU/mg) to the activity of the commercially produced standard protease procured from Novo Nordisk, Denmark (30.35 APU/mg). Hence, the proteolytic activity produced by this isolate was further investigated in batch and fed-batch process. Sucrose was the best carbon source for the production of protease activity by that isolate. Different organic nitrogen sources (casein, peptone and beef extract) at 1% (w/v) with varying levels of sucrose (1% - 4% w/v) initially repress enzyme synthesis. The duration and extent of repression decreased with increased concentration of sucrose. Maximum protease activity was found in basal medium with 4% (w/v) sucrose and 1% (w/v) yeast extract. Yeast-extract was thought to be an inducer of enzyme synthesis. Further, the basal medium was unique with respect to the enzyme production, as protease production was growth associated with the peak enzyme production being detected at the time of maximum growth. Interestingly, a rise in 34.2% (104.86 APU/ml) of protease activity was detected at incubation temperature of 50℃ and when culture filtrate was assayed at 60℃, signifying a high temperature stability of the produced protease by this isolate. Additional studies on the enzyme characterization were resulted in recognition of highly significant properties of the activity towards casein at pH 9.0 and stability at high temperature with retention of 96% the enzyme activity at 60℃. The parametric study under feed intervals had enabled improvement in the maximum protease activities attainable from batch cultures in excess of 21.78% and 26.32% via two feeding strategies. A small continual increase in enzyme activity (132.46 APU/ml during 24 h - 120 h) and enhancement in protease production in excess of 36.84% was observed by fed-batch process than the batch experiment.展开更多
An on-line scheduling algorithm to maximize gross profit of penicillin fed-batch fermentation is proposed. According to the on-line classification method, fed-batch fermentation batches are classified into three categ...An on-line scheduling algorithm to maximize gross profit of penicillin fed-batch fermentation is proposed. According to the on-line classification method, fed-batch fermentation batches are classified into three categories. Using the scheduling strategy, the optimal termination sequence of batches is obtained. Pseudo on-line simulations for testing the proposed algorithm with the data from industrial scale penicillin fermentation are carried out.展开更多
Bioreactor operation requires continuous monitoring of fermentation parameters and real-time control over bioreactor devices. Remote monitoring and control of the bioreactor＇s computer via the Internet avoids the nec...Bioreactor operation requires continuous monitoring of fermentation parameters and real-time control over bioreactor devices. Remote monitoring and control of the bioreactor＇s computer via the Internet avoids the necessity of personnel being continually onsite during operation. A two liter Sartorius-stedim Biostat~ A Plus fermentation system was networked and interfaced with the commercial software from GoToMyPC to allow remote control of the fermentation system utilizing the internet. The fermentation vessel was equipped with hardware calibrated for monitoring and controlling culture parameters during experimentations. The uniform resource locator controlled night-vision web camera allowed continuous monitoring of the glass fermentation vessel during the day and at night. The main window screen of the laboratory computer can be securely accessed from any portable device （i.e. laptop） capable of establishing an Internet connection and executing the commercial software from GoToMyPC. The secured internet protocol address, provided by GoToMyPC, assures that the system can only be controlled by authorized users who have been given access to the account. This interface permits the remote control of the Biostat A Plus fermentation system and possibly other automated or potentially automated culturing systems at the convenience of the user（s）.展开更多
Violacein,a therapeutic pigment synthesised naturally in specific bacterial systems,is commercially unexplored owing to low titers.The current study aimed to formulate a suitable medium and to develop a fed-batch stra...Violacein,a therapeutic pigment synthesised naturally in specific bacterial systems,is commercially unexplored owing to low titers.The current study aimed to formulate a suitable medium and to develop a fed-batch strategy for improving the violacein productivity in a natural producer-Chromobacterium violaceum MTCC2656.The carbon and the nitrogen sources were extensively screened and their levels were optimised for maximal violacein production.The micronutrients in the medium were subjected to a two-level statistical optimisation using design of experiment approach and the final media formulation was validated.As a fed-batch approach,a combination of pulse feeding of glucose and tryptophan with optimised broth harvest of 60%(v/v)was attempted that achieved a titer of 1046±16 mg/L with productivity of 26.12±0.64 mg/L/h in each progressive cycle of fed-batch.The strategic sequential step of media formulation and fed-batch fermentation improved the violacein titer by~5 folds.Kinetic modelling was used to understand the enhancement in fermentation performance in both the batch and fed-batch processes.The findings from the current study would enable to understand and correlate the patterns of substrate uptake and violacein formation to establish strategies for enhancing overall productivity in the fermentation processes.展开更多
文摘[Objective] This study was to realize high-density culture of Chlorella sp. as well as the culture with high lipid yield. [Method] Through analyzing the growth curve of Chlorella sp. cells, dextrose consumption curve, change curves of pH and dissolved oxygen, a fed-batch culture was conducted, followed by a nitrogen-deficient culture aiming at accumulating the lipids in Chlorella sp. cells when a high density of Chlorella sp. cells was obtained. [Result] After four batches of feeding were pro- vided, the biomass of Chlorella sp. reached up to 65.25 g/L, and the lipid content increased from 42.75% to 63.82% in Chlorella sp. cells, with the yield of 43.37 g/L in the following 12 hours of nitrogen-deficient culture. [Conclusion] Reasonable fed- batch can significantly improve the biomass of Chlorella sp., and the nitrogen-defi- cient culture further raises the lipid yield of Chlorella sp.
基金Supported by the National Science and Technology Key Funds (2003DA901A32)the National Nature Science Foundation (No. 20476085).
文摘Recombinant E. coli BL 21 was cultivated in high cell density to produce human-like collagen. The effects of the feeding of nitrogen source, controlled by an auto on/off-feeding mode with two different cycles of 0.5min and 4min intervals, oxygen-enrichment methods and inducement strength on the cell yield and human-like collagen production were investigated. The studies showed that nitrogen source feeding in fast cycle could result in higher human-like collagen production than that in slow cycle; and the feedback regulation of glucose, increase of the pressure of fermentation bioreactor, and supply of oxygen-enriched air could all increase cell yield and human-like collagen production. The effects of inducement strength on protein expression were found important. When OD600 reached 90-100, the cultivation temperature was increased to 42℃ to begin induction for 2-3 h, and then shifted to 39℃ for 5-6h induction, the cell density and human-like collagen production could reach 96g·L-1 [DCW (dry cell mass)] and 19.8% (g·g-1 DCW) respectively.
文摘The production of butyric acid by Clostridium butyricum ZJUCB at various pH values was investigated. In order to study the effect of pH on cell growth, butyric acid biosynthesis and reducing sugar consumption, different cultivation pH values ranging from 6.0 to 7.5 were evaluated in 5-L bioreactor. In controlled pH batch fermentation, the optimum pH for cell growth and butyric acid production was 6.5 with a cell yield of 3.65 g/L and butyric acid yield of 12.25 g/L. Based on these results, this study then compared batch and fed-batch fermentation of butyric acid production at pH 6.5. Maximum value (16.74 g/L) of butyric acid concentration was obtained in fed-batch fermentation compared to 12.25 g/L in batch fermentation. It was concluded that culti- vation under fed-batch fermentation mode could enhance butyric acid production significantly (P<0.01) by C. butyricum ZJUCB.
基金Supported by the National Natural Science Foundation of China （20676013）
文摘An iterative optimization strategy for fed-batch fermentation process is presented by combining a run-to-run optimization with swarm energy conservation particle swarm optimization (SEC-PSO). SEC-PSO, which is designed with the concept of energy conservation, can solve the problem of premature convergence frequently appeared in standard PSO algorithm by partitioning its population into several sub-swarms according to the energy of the swarm and is used in the optimization strategy for parameter identification and operation condition optimization. The run-to-run optimization exploits the repetitive nature of fed-batch processes in order to deal with the optimal problems of fed-batch fermentation process with inaccurate process model and unsteady process state. The kinetic model parameters, used in the operation condition optimization of the next run, are adjusted by calculating time-series data obtained from real fed-batch process in the run-to-run optimization. The simulation results show that the strategy can adjust its kinetic model dynamically and overcome the instability of fed-batch process effectively. Run-to-run strategy with SEC-PSO provides an effective method for optimization of fed-batch fermentation process.
基金Partially suppored by a grant for the U K DTI- China MOST Collaborative Research
文摘L-(+)-Lactic acid production from corncob hydrolysate as a cheap carbohydrate source by fed-batch fermentation of Rhizopus oryzaeHZS6 was studied. After 96 h of fermentation in a 5 L fermentor, the final concentration of ammonium L-(+)-lactate, average productivity(based on initial xylose concentration) and maximum dry cell weight were 132.4 g/L, 1.38 g/(L·h), and 8.9 g/L respectively. The optical purity of L-(+)-lactate was 98.8%.
基金Supported by the National Natural Science Foundation of China （20306026 and 20876141） and the National Basic Research program of China （2007CB707805）.
文摘5-Aminolevulinic acid （ALA） is a common precursor for tetrapyrrole compounds in all kinds of organ isms and has wide applications in agriculture and medicines. In this study, a new strategy, i.e. short-term dissolved oxygen （DO） shock during aerobic fermentation, was introduced to produce 5-aminolevulinic acid with a recombi-nant E. coli. Effects of duration time of DO shock operation on plasmid concentration, intracellular ALA synthase （ALAS） activity and ALA production were investigated in Erlenmeyer shake flasks. The results indicated that both ALAS activity and ALA yield were enhanced in an anaerobic operation of 45 rain in the early exponential phase during fermentation, while they decreased when the anaerobic operation time was further increased to 60 rain. The DO shock protocol was confirmed with the fed-batch fermentation in a 15 L fermenter and the ALA production achieved 9.4 g.L-1 （72 mmol.L-1）, which is the highest yield in the fermentation broth reported up to now.
基金Supported by the National Natural Science Foundation of China,No.29476248 and Trans-Century Training Program Foundation for the Talents by the State Education Commission.
文摘In this paper,a novel fuzzy neural network model,in which an adjustable fuzzy sub-space was designed by uniform design,has been established and used in fed-batch yeast fermentationas an example.A brand-new optimization sub-network with special structure has been built andgenetic algorithm,guaranteeing the optimization in overall space,is introduced for the feed rateoptimization.On the basis of the model network,the optimal substrate concentration and theoptimal amount of fed-batch at different periods have been studied,aided with the optimizationnetwork and the genetic algorithm separately.The above results can be used as a basis for theestablishment of a fuzzy neural network controller.
基金Supported by the National Natural Science Foundation of China (20476007, 20676013)
文摘In this study, Saccharomyces cerevisiae (baker's yeast) was produced in a fed-batch bioreactor at the optimal dissolved oxygen concentration (DOC) and growth medium temperature. However, it is very difficult to control the DOC using conventional controllers because of the poorly understood and constantly changing dynamics of the bioprocess. A generalized predictive controller (GPC) based on a nonlinear autoregressive integrated moving average exogenous (NARIMAX) model is presented to stabilize the DOC by manipulation of air flow rate. The NARIMAX model is built by an improved recursive least-squares support vector machine, which is trained by an in-place computation scheme and avoids the computation of the inverse of a large matrix and memory reallocation. The proposed nonlinear GPC algorithm requires little preliminary knowledge of the fermentation process, and directly obtains the nonlinear model in matrix form by using iterative multiple modeling instead of linearization at each sampling period. By application of an on-line bioreactor control, experimental results demonstrate the robustness, effectiveness and advantages of the new controller.
文摘An alkalophilc and thermophilic Bacillus sp. BHA that produced a thermostable alkaline protease was isolated from decaying protein substrates. The isolate was found to grow in pH range 7 - 11 with an optimum pH 9.0 and temperature up to 55℃. The activity of alkaline protease of Bacillus sp. BHA (68.98 APU/ml) was found higher than the standard strains of Bacillus amyloliquefaciens MTCC 610 (8.98 APU/ml) and Bacillus subtilis MTCC 8349 (12.14 APU/ml, used in this study, and was comparable (68.98 APU/ml, equivalent to 30.38 APU/mg) to the activity of the commercially produced standard protease procured from Novo Nordisk, Denmark (30.35 APU/mg). Hence, the proteolytic activity produced by this isolate was further investigated in batch and fed-batch process. Sucrose was the best carbon source for the production of protease activity by that isolate. Different organic nitrogen sources (casein, peptone and beef extract) at 1% (w/v) with varying levels of sucrose (1% - 4% w/v) initially repress enzyme synthesis. The duration and extent of repression decreased with increased concentration of sucrose. Maximum protease activity was found in basal medium with 4% (w/v) sucrose and 1% (w/v) yeast extract. Yeast-extract was thought to be an inducer of enzyme synthesis. Further, the basal medium was unique with respect to the enzyme production, as protease production was growth associated with the peak enzyme production being detected at the time of maximum growth. Interestingly, a rise in 34.2% (104.86 APU/ml) of protease activity was detected at incubation temperature of 50℃ and when culture filtrate was assayed at 60℃, signifying a high temperature stability of the produced protease by this isolate. Additional studies on the enzyme characterization were resulted in recognition of highly significant properties of the activity towards casein at pH 9.0 and stability at high temperature with retention of 96% the enzyme activity at 60℃. The parametric study under feed intervals had enabled improvement in the maximum protease activities attainable from batch cultures in excess of 21.78% and 26.32% via two feeding strategies. A small continual increase in enzyme activity (132.46 APU/ml during 24 h - 120 h) and enhancement in protease production in excess of 36.84% was observed by fed-batch process than the batch experiment.
基金supported by the Open Project Program,State key Laboratory of Bioreactor Engineering/ECUSTthe Natural Science Foundation of China(No.60174024).
文摘An on-line scheduling algorithm to maximize gross profit of penicillin fed-batch fermentation is proposed. According to the on-line classification method, fed-batch fermentation batches are classified into three categories. Using the scheduling strategy, the optimal termination sequence of batches is obtained. Pseudo on-line simulations for testing the proposed algorithm with the data from industrial scale penicillin fermentation are carried out.
文摘Bioreactor operation requires continuous monitoring of fermentation parameters and real-time control over bioreactor devices. Remote monitoring and control of the bioreactor＇s computer via the Internet avoids the necessity of personnel being continually onsite during operation. A two liter Sartorius-stedim Biostat~ A Plus fermentation system was networked and interfaced with the commercial software from GoToMyPC to allow remote control of the fermentation system utilizing the internet. The fermentation vessel was equipped with hardware calibrated for monitoring and controlling culture parameters during experimentations. The uniform resource locator controlled night-vision web camera allowed continuous monitoring of the glass fermentation vessel during the day and at night. The main window screen of the laboratory computer can be securely accessed from any portable device （i.e. laptop） capable of establishing an Internet connection and executing the commercial software from GoToMyPC. The secured internet protocol address, provided by GoToMyPC, assures that the system can only be controlled by authorized users who have been given access to the account. This interface permits the remote control of the Biostat A Plus fermentation system and possibly other automated or potentially automated culturing systems at the convenience of the user（s）.
基金supported by the National Natural Science Foundation of China(No.21076148 and No.31270087)Plan of Tianjin Science and Technology Support(11ZCKFSY0100)
基金funded by the All-India Council of Technical Education,New Delhi,India。
文摘Violacein,a therapeutic pigment synthesised naturally in specific bacterial systems,is commercially unexplored owing to low titers.The current study aimed to formulate a suitable medium and to develop a fed-batch strategy for improving the violacein productivity in a natural producer-Chromobacterium violaceum MTCC2656.The carbon and the nitrogen sources were extensively screened and their levels were optimised for maximal violacein production.The micronutrients in the medium were subjected to a two-level statistical optimisation using design of experiment approach and the final media formulation was validated.As a fed-batch approach,a combination of pulse feeding of glucose and tryptophan with optimised broth harvest of 60%(v/v)was attempted that achieved a titer of 1046±16 mg/L with productivity of 26.12±0.64 mg/L/h in each progressive cycle of fed-batch.The strategic sequential step of media formulation and fed-batch fermentation improved the violacein titer by~5 folds.Kinetic modelling was used to understand the enhancement in fermentation performance in both the batch and fed-batch processes.The findings from the current study would enable to understand and correlate the patterns of substrate uptake and violacein formation to establish strategies for enhancing overall productivity in the fermentation processes.
