Fibrous Bed Bioreactor Research Articles

Thermophilic combined bioproduction of ethanol and hydrogen utilizing sugarcane bagasse

Research addressing investigation of environment-friendly and efficient fuel reservoirs substituting depleting fossil fuels is at-work worldwide. For this purpose, it is of ultimate requirement to establish cost-effective biofuel fermentative processes using renewable lignocellulosic bio-wastes as feedstock. The present investigation compacts the employment of Clostridium thermocellum DSMZ1313, for bench-scale co-production of bioethanol and biohydrogen (bioH2) under non-aseptic thermophilic conditions. In our previous study, the fermentation kinetics have been studied aseptically in bench-scale stirred-tank bioreactor with same fermentative bacterium under optimized levels of the key fermentation factors employing sugarcane bagasse (SCB) in the fermentation medium. In the current investigation, for co-production of bioethanol and bioH2, batch fermentation was carried as free cell approach in subsequent bioreactor’s modifications that resulted in enhanced biofuels’ yields aseptically. These include the incorporation of immobilized cells fibrous bed bioreactor (FBB) that caused 20.465% enhanced ethanol and 27.27% higher level of hydrogen, followed by substrate addition (fed-batch fermentation) resulted in 61.79% increased ethanol and 74.074% better hydrogen production. Selective stripping of the inhibitory metabolites by in-situ gas-stripping was also performed that resulted in 94.58% increased ethanol and 105.38% elevated hydrogen level. Finally, under non-aseptic thermophilic fermentation conditions, 16.07% reduced ethanol whereas 14.7% elevated hydrogen levels were obtained in comparison with aseptic fermentation conditions. The simultaneous formation of bioethanol and bioH2 by SCB usage as biofuels’ feedstock from C. thermocellum-DSMZ1313 under non-aseptic settings has appeared a potentially sustainable recourse to produce environmentally friendly energy sources. To overcome the non-sterile fermentation processes’ inefficiencies, further studies might prove useful for active large scale deployment of waste biomass-to-biofuel.

Food waste from various origins as feedstocks for recombinant protein production by Yarrowia lipolytica using in situ fibrous bed bioreactor

Recombinant proteins (rProts) hold diverse applications and exhibit promising market potential. However, the impediment to their industrial production lies in the high associated costs, notably those related to the medium components. This study focuses on the heterologous production of the lipase B (CalB) from Candida antarctica, used as a case study, from food waste by Yarrowia lipolytica. Various food waste hydrolysates (FWHs) were assessed, and Chinese restaurant FWH emerged as the most suitable feedstock for both cell growth and rProt production. Employing a response surface methodology, the culture conditions and medium compositions were optimised, resulting in a 139.4% increase in lipase production in shake flasks, yielding a titre of 6.32 × 103 U/mL and a productivity of 65.83 U/(mL·h). The lipase titre and productivity achieved in a 2.5-L stirred-tank bioreactor were subsequently increased by 81.0% and 122.8%, respectively. In an in situ fibrous bed bioreactor (isFBB) with cells immobilised on sugarcane bagasse, a pulsed fed-batch strategy led to further enhancement, reaching a lipase productivity of 265.63 U/(mL⋅h). This represents an increase of 80.7% compared to a similar process using glycerol as a feedstock. This study demonstrates clearly that FWHs can be used for rProt production using Y.lipolytica as a cell factory.

Facile and green synthesis of reduced graphene oxide/loofah sponge for Streptomyces albulus immobilization and ε-poly-l-lysine production

Facile and green fabrication of reduced graphene oxide on loofah sponge (rGOLS) carrier was applied for cell immobilization of ε-Poly-l-lysine (ε-PL) production. Due to surface properties including large specific surface area, high polarity, and low interaction energy, rGOLS-1 was employed as the optimum rGOLS to enhance immobilization of Streptomyces albulus. Compared with raw LS, batch experiments showed rGOLS-1 facilitated superior cell vitality for ε-PL production due to the presence of reduced graphene oxide. In the sequential fed-batch cultivation of Streptomyces albulus using rGOLS-1 with an aerobic plant fibrous-bed bioreactor (rGOLS-1-AFPB), the maximum ε-PL concentration and productivity reached to 39.2 ± 0.63 g/L and 0.48 g/L/h. The cells immobilized in rGOLS-1 with high vitality and ε-PL production efficiency were reused six times over a period of 624 h. This research afforded an effective approach to enhance the fermentation performance of immobilized cells with the design of an advanced immobilization carrier.

Isopropanol-butanol-ethanol production by cell-immobilized vacuum fermentation

The Isopropanol-Butanol-Ethanol productivity by solventogenic clostridia can increase when cells are immobilized on low-cost, renewable fibrous materials; however, butanol inhibition imposes the need for dilute sugar solutions (less than40 g/L). To alleviate this problem, the in-situ vacuum product recovery technique was applied to recover IBE in repeated-batch cultivation of Clostridium beijerinckii DSM 6423 immobilized on sugarcane bagasse. Five repeated batch cycles were conducted in a 7-L bioreactor containing P2 medium (∼60 g/L glucose) and bagasse packed in 3D-printed concentric annular baskets. In three cycles, glucose was consumed by 86% on average, the IBE productivity was 0.35 g/L∙h or 30% and 17% higher relative to free- and immobilized (without vacuum)-cell cultures. Notably, the product stream contained 45 g/L IBE. However, the fermentation was unsatisfactory in two cycles. Finally, by inserting a fibrous bed with hollow annuli in a vacuum fermentation, this work introduces the concept of an internal-loop boiling-driven fibrous-bed bioreactor.

Enhancing the recombinant protein productivity of Yarrowia lipolytica using in situ fibrous bed bioreactor

In this study, the ability of Yarrowia lipolytica to produce the recombinant lipase CalB from Candida antarctica, used as a model protein has been compared across different bioreactor processes using glycerol, a byproduct from the biodiesel industry as the main carbon source. Batch, pulsed fed-batch (PFB), and continuous fed-batch (CFB) strategies were first compared using classical stirred tank (STR) bioreactors in terms of biomass production, carbon source uptake, and lipase production. Additionally, an in situ fibrous bed bioreactor (isFBB) was developed using sugarcane bagasse as a cell immobilization support. The maximum lipase titer achieved using the isFBB culture mode was 38%, 33%, and 49% higher than those obtained using the batch, PFB, and CFB cultures, respectively. The lipase productivity in isFBB mode (142U/mL/h) was 1.4-fold higher than that obtained using batch free cell cultures. These results highlight that isFBB is an efficient technology for the production of recombinant enzymes.

Isolation and characterization of a newly identified Clostridium butyricum strain SCUT343-4 for 1,3-propanediol production.

A novel 1,3-propanediol (1,3-PDO) producing strain was isolated and identified as Clostridium butyricum with respect to its morphological and physiological characteristics, as well as 16S rDNA. The results of substrates test and stress tolerance indicated that C. butyricum SCUT343-4 could produce 1,3-PDO efficiently from glycerol. The optimal fermentation conditions were determined to be 5g/L yeast extract at 37°C and pH 6.5. To fully evaluate its 1,3-PDO production capacity, different cultivation strategies have been implemented. The highest 1,3-PDO concentration obtained for batch and fed-batch fermentation were 51.64 and 61.30g/L, respectively. Immobilized cell fermentation in fibrous-bed bioreactor was also performed, and the concentration of 1,3-PDO further increased to 86g/L with a yield of 0.52g/g. In addition, the 1,3-PDO productivity reached 4.20g/L h, which is the highest level reported for C. butyricum, demonstrating the potential of C. butyricum SCUT343-4 for 1,3-PDO production from glycerol.

Role of microbubbles coupling fibrous-bed bioreactor in butyric acid production by Clostridium tyrobutyricum using Brewer’s spent grain as feedstock

Microbubbles coupling fermentation is an efficient technology for improving oxygen transfer to aerobic microorganisms in a bioreactor containing an air sparger. However, the effect of this technology on anaerobic Clostridium tyrobutyricum, whose morphology develops viscous broth rheology and lumps impeding the nutrient mass transfer, has not been reported yet. Therefore, we evaluated the role of microbubbles coupling fibrous-bed bioreactor (MBFBB) on butyrate production by immobilized Clostridium tyrobutyricum cells obtained from renewable feedstock brewer’s spent grain (BSG). Compared with the conventional fibrous-bed bioreactor (FBB) fermentation, two-fold shorter fermentation time and two-fold higher butyrate productivity were achieved by MBFBB-immobilized C. tyrobutyricum cells. Furthermore, long-term stability and reliability for butyrate production were obtained in ten cycle process using BSG hydrolysate. Finally, fed-batch fermentation using BSG hydrolysate produced a high butyrate titer of 43.68 g/L, with a significantly higher selectivity of 11.67, reducing the production cost of downstream processing. The results also indicated that butyrate productivity of 5.40 g/(L h were successfully obtained in BSG medium, thereby representing the highest productivity reported to date in C. tyrobutyricum from low-cost feedstocks. To our knowledge, this is also the first report related to microbial production of butyrate from BSG. Based on techno-economic analysis, it was demonstrated that the MBFBB-based fermentation process with BSG, was robust, economic and should underpin the future development of commercial biobutyrate production.

Energy-efficient butanol production by Clostridium acetobutylicum with histidine kinase knockouts to improve strain tolerance and process robustness

Engineering histidine kinases in C. acetobutylicum enhanced cell viability and solventogenesis in ABE fermentation and enabled robust and energy-efficient butanol production.

Efficient production of butyric acid by Clostridium tyrobutyricum immobilized in an internal fibrous bed bioreactor (IFBB)

An internal fibrous bed bioreactor (IFBB) was constructed in an anaerobic stirred fermenter for butyric acid production by immobilized Clostridium tyrobutyricum ATCC 25755. In the IFBB system, porous cotton terry cloth was used as a fibrous matrix and the matrix was fixed on agitation shaft and inside baffles. Firstly, free-cell fermentations with batch and fed-batch modes were carried out under pH 6.0. The yield and butyric acid/acetic acid (B/A) ratio reached 0.387 g/g and 4.70 g/g in batch mode, and 0.426 g/g and 8.09 g/g in fed-batch mode, respectively. When implementing fed-batch fermentation with immobilized cells in IFBB, the concentration and productivity of butyric acid reached 45.38 g/L and 0.630 g/L/h, respectively, which were enhanced by 10 % in comparison with the free-cell fed-batch fermentation. Furthermore, a high yield of 0.440 g/g and B/A ratio of 12.75 g/g were also achieved in IFBB with fed-batch mode. Finally, the productivity of butyric acid could be further elevated to ∼0.90 g/L/h with repeated-batch in IFBB, while it was only 0.444 g/L/h in free-cell batch fermentation. These results obtained might show the potential for efficient butyric acid production in the IFBB system from renewable resources.

Production of n-butanol from cassava bagasse hydrolysate by engineered Clostridium tyrobutyricum overexpressing adhE2: Kinetics and cost analysis

The production of biofuels such as butanol is usually limited by the availability of inexpensive raw materials and high substrate cost. Using food crops as feedstock in the biorefinery industry has been criticized for its competition with food supply, causing food shortage and increased food prices. In this study, cassava bagasse as an abundant, renewable, and inexpensive byproduct from the cassava starch industry was used for n-butanol production. Cassava bagasse hydrolysate containing mainly glucose was obtained after treatments with dilute acid and enzymes (glucoamylases and cellulases) and then supplemented with corn steep liquor for use as substrate in repeated-batch fermentation with engineered Clostridium tyrobutyricum CtΔack-adhE2 in a fibrous-bed bioreactor. Stable butanol production with high titer (>15.0 g/L), yield (>0.30 g/g), and productivity (~0.3 g/L∙h) was achieved, demonstrating the feasibility of an economically competitive process for n-butanol production from cassava bagasse for industrial application.

Potential of hydrogen production from sugarcane juice by Ethanoligenens harbinense Yuan-3

Inexpensive biomass is an ideal substrate for low-cost production of biohydrogen, but little was known about the biomass utilization performance of the ethanol-type hydrogen-producing bacteria Ethanoligenens harbinense. This study investigated the feasibility and performance of biohydrogen production by Ethanoligenens harbinense from sugarcane juice (SJ). Medium and process were firstly optimized with SJ as carbon source and the highest hydrogen yield was obtained when SJ was hydrolyzed by acid, and the medium contained 10 g/L sugar and 3 g/L yeast extract with pH at 6.0. Compared to batch fermentation, fed-batch fermentation with cell recycle showed significant improvement in hydrogen productivity and yield. The maximum productivity reached 5.22 mmol-H2/L•h, 201.7% higher than that of the single batch (1.73 mmol-H2/L•h). The highest H2 yield was 2.23 mol-H2/mol-hexose, 65.2% higher than that of single batch fermentation. Cell immobilization with the fibrous bed bioreactor (FBB) also improved hydrogen productivity to 2.6 times that of batch fermentation with free cells. This study demonstrated the feasibility of hydrogen production from SJ with Ethanoligens sp., and a great improvement in hydrogen production was obtained by recycling cells and immobilizing cells in a fibrous bed bioreactor.

N-Butanol production from lignocellulosic biomass hydrolysates without detoxification by Clostridium tyrobutyricum Δack-adhE2 in a fibrous-bed bioreactor

Acetone-butanol-ethanol fermentation suffers from high substrate cost and low butanol titer and yield. In this study, engineered Clostridium tyrobutyricum CtΔack-adhE2 immobilized in a fibrous-bed bioreactor was used for butanol production from glucose and xylose present in the hydrolysates of low-cost lignocellulosic biomass including corn fiber, cotton stalk, soybean hull, and sugarcane bagasse. The biomass hydrolysates obtained after acid pretreatment and enzymatic hydrolysis were supplemented with corn steep liquor and used in repeated-batch fermentations. Butanol production with high titer (∼15 g/L), yield (∼0.3 g/g), and productivity (∼0.3 g/L∙h) was obtained from cotton stalk, soybean hull, and sugarcane bagasse hydrolysates, while corn fiber hydrolysate with higher inhibitor contents gave somewhat inferior results. The fermentation process was stable for long-term operation without any noticeable degeneration, demonstrating its potential for industrial application. A techno-economic analysis showed that n-butanol could be produced from lignocellulosic biomass using this novel fermentation process at ∼$2.5/gal for biofuel application.

Efficient succinic acid production using a biochar-treated textile waste hydrolysate in an in situ fibrous bed bioreactor

Textile waste contains biodegradable fraction which can be used as an alternative feedstock for succinic acid production. The feasibility of bioconversion of biochar-treated textile waste hydrolysate into succinic acid was evaluated. A substrate loading of 9% was applied to hydrolyze mixed textile waste and the resultant glucose-rich hydrolysate with dyestuff was collected after enzymatic hydrolysis. Biochar from different pyrolysis temperatures (400–700 °C) and different dosages of the selected biochar (1–5%) were applied to remove colorants. The results showed that biochar had a good performance at a dosage of 2 w/w % on removal of colorant inhibitors. There was no negative effect observed during the subsequent fermentation. After optimization of fermentation media in shake flasks, the resultant succinic acid titer reached 19.6 g/L with a SA yield of 0.76 g/g, supplemented with 30 g/L tryptone. The in situ fibrous bed bioreactor (isFBB) can further improve SA titer, up to 28.8 g/L corresponding to a yield of 0.61 g/g without pH control. Furthermore, a shorter lag phase during succinic acid fermentation in isFBB was observed. It can be concluded that a novel textile waste-based biorefinery approach for succinic acid production aided by biochar sorption was successfully developed.

Efficient metabolic evolution of engineered Yarrowia lipolytica for succinic acid production using a glucose-based medium in an in situ fibrous bioreactor under low-pH condition

BackgroundAlkali used for pH control during fermentation and acidification for downstream recovery of succinic acid (SA) are the two largest cost contributors for bio-based SA production. To promote the commercialization process of fermentative SA, the development of industrially important microorganisms that can tolerate low pH has emerged as a crucial issue.ResultsIn this study, an in situ fibrous bed bioreactor (isFBB) was employed for the metabolic evolution for selection of Y. lipolytica strain that can produce SA at low pH using glucose-based medium. An evolved strain named Y. lipolytica PSA3.0 that could produce SA with a titer of 19.3 g/L, productivity of 0.52 g/L/h, and yield of 0.29 g/g at pH 3.0 from YPD was achieved. The enzyme activity analysis demonstrated that the pathway from pyruvate to acetate was partially blocked in Y. lipolytica PSA3.0 after the evolution, which is beneficial to cell growth and SA production at low pH. When free-cell batch fermentations were performed using the parent and evolved strains separately, the evolved strain PSA3.0 produced 18.4 g/L SA with a yield of 0.23 g/g at pH 3.0. Although these values were lower than that obtained by the parent strain PSA02004 at its optimal pH 6.0, which were 25.2 g/L and 0.31 g/g, respectively, they were 4.8 and 4.6 times higher than that achieved by PSA02004 at pH 3.0. By fed-batch fermentation, the resultant SA titer of 76.8 g/L was obtained, which is the highest value that ever achieved from glucose-based medium at low pH, to date. When using mixed food waste (MFW) hydrolysate as substrate, 18.9 g/L SA was produced with an SA yield of 0.38 g/g, which demonstrates the feasibility of using low-cost glucose-based hydrolysate for SA production by Y. lipolytica in a low-pH environment.ConclusionsThis study presents an effective and efficient strategy for the evolution of Y. lipolytica for SA production under low-pH condition for the first time. The isFBB was demonstrated to improve the metabolic evolution efficiency of Y. lipolytica to the acidic condition. Moreover, the acetate accumulation was found to be the major reason for the inhibition of SA production at low pH by Y. lipolytica, which suggested the direction for further metabolic modification of the strain for improved SA production. Furthermore, the evolved strain Y. lipolytica PSA3.0 was demonstrated to utilize glucose-rich hydrolysate from MFW for fermentative SA production at low pH. Similarly, Y. lipolytica PSA3.0 is expected to utilize the glucose-rich hydrolysate generated from other carbohydrate-rich waste streams for SA production. This study paves the way for the commercialization of bio-based SA and contributes to the sustainable development of a green economy.

Production of butyric acid from acid hydrolysate of corn husk in fermentation by Clostridium tyrobutyricum: kinetics and process economic analysis

BackgroundButyric acid is an important chemical currently produced from petrochemical feedstocks. Its production from renewable, low-cost biomass in fermentation has attracted large attention in recent years. In this study, the feasibility of corn husk, an abundant agricultural residue, for butyric acid production by using Clostridium tyrobutyricum immobilized in a fibrous bed bioreactor (FBB) was evaluated.ResultsHydrolysis of corn husk (10% solid loading) with 0.4 M H2SO4 at 110 °C for 6 h resulted in a hydrolysate containing ~ 50 g/L total reducing sugars (glucose:xylose = 1.3:1.0). The hydrolysate was used for butyric acid fermentation by C. tyrobutyricum in a FBB, which gave 42.6 and 53.0% higher butyric acid production from glucose and xylose, respectively, compared to free-cell fermentations. Fermentation with glucose and xylose mixture (1:1) produced 50.37 ± 0.04 g L−1 butyric acid with a yield of 0.38 ± 0.02 g g−1 and productivity of 0.34 ± 0.03 g L−1 h−1. Batch fermentation with corn husk hydrolysate produced 21.80 g L−1 butyric acid with a yield of 0.39 g g−1, comparable to those from glucose. Repeated-batch fermentations consistently produced 20.75 ± 0.65 g L−1 butyric acid with an average yield of 0.39 ± 0.02 g g−1 in three consecutive batches. An extractive fermentation process can be used to produce, separate, and concentrate butyric acid to > 30% (w/v) sodium butyrate at an economically attractive cost for application as an animal feed supplement.ConclusionA high concentration of total reducing sugars at ~ 50% (w/w) yield was obtained from corn husk after acid hydrolysis. Stable butyric acid production from corn husk hydrolysate was achieved in repeated-batch fermentation with C. tyrobutyricum immobilized in a FBB, demonstrating that corn husk can be used as an economical substrate for butyric acid production.

Hydrolysis of fruit and vegetable waste for efficient succinic acid production with engineered Yarrowia lipolytica

Abstract Bio-based production of succinic acid (SA) is regarded to supplement the petro-based processing for environmental reasons. However, in order to enhance the industrial competitiveness of bio-based SA, it is necessary to develop a method that is cost-effective, environmental friendly and with high efficiency. In this study, the utilization of fruit and vegetable waste (FVW) hydrolysate for SA production via engineered Yarrowia lipolytica was reported for the first time. In the study, the hydrolysis conditions were firstly optimized and the final glucose concentration of 56.7 g L−1 and a yield of 0.46 g g−1 were obtained with 2% glucoamylase, 1% cellulase, 2% hemicellulase and 0.25% pectinase at pH 5.0 and 55 °C. Then, the feasibility of using the hydrolysate for SA production via Y. lipolytica PSA02004 was demonstrated. SA titer and yield at 43.1 g L−1 and 0.46 g g−1, respectively were obtained using 100 g L−1 glucose-containing FVW hydrolysate and 4% corn steep liquor (CSL) as fermentation medium by free cell batch fermentation. Finally, in situ fibrous bed bioreactor (isFBB) and fed batch fermentation were demonstrated to be effective approach for improved SA production, from which SA productivity of 0.69 g L−1 h−1 and the resultant SA concentration of 140.6 g L−1 were achieved, respectively. In summary, this study achieved the anticipated target of turning waste into value-added products with high efficiency and cost-effectiveness.

Repeated-batch fermentation by immobilization of Clostridium beijerinckii NCIMB 8052 in a fibrous bed bioreactor for ABE (acetone-butanol-ethanol) production

In this study, a fibrous bed bioreactor (FBB) system was used to produce ABE (acetone-butanol-ethanol) by immobilized cells of Clostridium beijerinckii NCIMB 8052. To obtain the maximum ABE productivity and yield and maintain the stability of a repeated-batch fermentation process in the FBB system, the optimal dilution rate (0.17 min−1) and initial glucose concentration (18 g/l) in the bioreactor were determined. When the repeated-batch fermentation was maintained at 11 cycles under the optimal conditions, an average of 4.9 g/l butanol (maximum 5.3 g/l) was achieved with an average solvent productivity of 1.0 g/l/h and a yield of 0.41 g/g (maxima of 1.1 g/l/h and 0.43 g/g, respectively). However, the solvent productivity and yield in batch fermentation of free cells were just 0.29 g/l/h and 0.36 g/g, respectively. In addition, scanning electron micrographs demonstrated that C. beijerinckii cells can be adsorbed onto a cotton towel and they exhibited a significantly different morphology compared to immobilized Clostridium acetobutylicum.

Comparative genomic analysis of Clostridium acetobutylicum for understanding the mutations contributing to enhanced butanol tolerance and production

Clostridium acetobutylicum JB200 is a hyper butanol tolerant and producing strain obtained from asporogenic C. acetobutylicum ATCC 55025 through mutagenesis and adaptation in a fibrous bed bioreactor. The complete genomes of both strains were sequenced by the Illumina Hiseq2000 technology and assembled using SOAPdenovo approach. Compared to the genomic sequence of the type strain ATCC 824, 143 single nucleotide polymorphisms (SNPs) and 17 insertion/deletion variations (InDels) were identified in the genome of ATCC 55025. Twenty-nine mutations were in genes involved in sporulation, solventogenesis and stress response. Compared to ATCC 55025, there were seven additional point mutations in the chromosome of JB200. Among them, a single-base deletion in cac3319 encoding an orphan histidine kinase caused protein C-terminal truncation. Disruption of this gene in ATCC 55025 and ATCC 824 resulted in significantly elevated butanol tolerance and production. This study provides genome-level information for the better understanding of solventogenic C. acetobutylicum in several key aspects of cell physiology and metabolism, which could help further metabolic engineering of Clostridium for butanol production.

Green and sustainable succinic acid production from crude glycerol by engineered Yarrowia lipolytica via agricultural residue based in situ fibrous bed bioreactor

In situ fibrous bed bioreactor (isFBB) for efficient succinic acid (SA) production by Yarrowia lipolytica was firstly developed in our former study. In this study, agricultural residues including wheat straw, corn stalk and sugarcane bagasse were investigated for the improvement of isFBB, and sugarcane bagasse was demonstrated to be the best immobilization material. With crude glycerol as the sole carbon source, optimization for isFBB batch fermentation was carried out. Under the optimal conditions of 20g sugarcane bagasse as immobilization material, 120gL−1 crude glycerol as carbon source and 4Lmin−1 of aeration rate, the resultant SA concentration was 53.6gL−1 with an average productivity of 1.45gL−1h−1 and a SA yield of 0.45gg−1. By feeding crude glycerol, SA titer up to 209.7gL−1 was obtained from fed batch fermentation, which was the highest value that ever reported.

Construction of fibrous bed bioreactor for enhanced succinic acid production using wastewater of dextran fermentation.

A new applicability of wastewater of dextran fermentation (WWDF) for biological production of succinic acid with A. succinogenes CCTCC M2012036 was reported in this work for the first time. Notably, K2CO3 was used instead of MgCO3 in the pH regulating process for operational feasibility and a cell immobilization methodology by attaching cells on cotton fibrous matrix was adopted for cell recycle. The initial sugar concentration as well as matrix usage was optimized by investigating the cell growth, succinic acid concentration and yield. A rotated fibrous bed bioreactor was designed and constructed in order to increase the total cell amount and facilitate mass transportation in the fermentation system, and an average succinic acid yield, concentration and productivity of 0.82g/g, 56.5g/L and 1.28g/L/h were realized in the repeated fed-batch fermentation, respectively. This research gave light to the optimization of succinic acid production towards a more cost-effective and operable direction.