Effect of organic solvents on enzymatic production of cyclodextrins from unliquefied corn starch in an attrition bioreactor

Cyclodextrins were produced from unliquefied corn starch in the presence of water-miscible organic solvents using cyclodextrin glycosyltransferase (CGTase) in an attrition bioreactor. The production yield was singnificatly increased by isopropanol and tertiary butanol, and maximum enhancement was observed to be about 40% by 5% tertiary butnol. Increase in the production of cyclodextrins by organic solvents seems to be due to the fact that organic solvents decreased the product inhibition of CGTase by forming an inclusion complex with cyclodextrins.     

Continuous production of cyclodextrins in an ultrafiltration membrane reactor,catalyzed by cyclodextrin glycosyltransferase from Bacillus circulans DF 9R

Cyclodextrins (CDs) are cyclic oligosaccharides of wide industrial application, whose synthesis is catalyzed by Cyclodextrin glycosyltransferase (CGTase) from starch. Here, CDs were produced using CGTase from Bacillus circulans DF 9R in continuous process and an ultrafiltration membrane reactor. The batch process was conducted as a control. This method allowed increasing the yield from 40 to 55.6% and the productivity from 26.1 to 99.5 mg of CD per unit of enzyme. The method also allowed obtaining a high‐purity product. The flow rate remained at 50% of its initial value after 24 h of process, improving the results described in the literature for starch hydrolysis processes. CGTase remained active throughout the process, which could be explained by the protective effect of the substrate and reaction products on CGTase stability. In addition, batch processes were developed using starches from different sources. We concluded that any of the starches studied could be used as substrate for CD production with similar yields and product specificity. © 2015 American Institute of Chemical Engineers Biotechnol. Prog., 31:695–699, 2015     

Enzymatic production of cyclodextrins from unliquefied corn starch in an attrition bioreactor

A novel enzymatic process for the production of cyclodextrins from unliquefied starch was developed. Cyclodextrins were produced in an attrition bioreactor in which simultaneous hydrolysis of starch and synthesis of cyclodextrins by cyclodextrin glycosyltransferase (CGTase) occur. The CGTase was obtained from isolated Bacillus sp. BE101, and maximum activity of the enzyme was observed at pH 6.0 and a temperature of 45 degrees C. The effect of milling media size and material on the performance of the attrition bioreactor was investigated, and operational parameters such as agitation speed, volume of milling media, ratio of enzyme to starch, and starch concentration were optimized. The production yield of cyclodextrins from unliquefied corn starch of 15% reached 35% at 24 h under optimized conditions. Energy consumption for the production of cyclodextrins in the attrition bioreactor system was estimated to be about 25% of that required for the liquefaction of starch in the conventional process.     

An ultrafiltration membrane bioreactor for the lipolysis of olive oil in reversed micellar media

The enzymatic hydrolysis of olive oil using Chromobacterium viscosum lipase B encapsulated in reversed micelles of dioctyl sodium sulfosuccinate (AOT) in isooctane was investigated in an ultrafiltration ceramic membrane reactor of tubular type, operating in a batch mode. Water concentration was found to be a critical parameter in the enzyme kinetics and hydrolysis yield of the reaction. The size of micelles, recirculation rate, and substrate concentration were found to be the major factors affecting the separation process. A correlation that enables the prediction of final conversion degrees in this bioreactor from the initial reaction conditions was established. (c) 1993 Wiley & Sons, Inc.     

Studies on enzymatic continuous production of cyclodextrins in an ultrafiltration membrane bioreactor

Studies on simultaneous hydrolysis of starch and synthesis of cyclodextrins by Thermo-aerobacter cyclodextrin glucosyltransferase were conducted in an ultrafiltration membrane bioreactor, allowing enzyme recovery and reduction of product inhibition. The influence of various reaction parameters like starch concentration, enzyme dosage and residence time on cyclodextrin composition was tested. A comparison of batch and continuous cyclodextrin production indicates that employing an ultrafiltration membrane bioreactor increases process efficiency.     

Biological hydrogen production using a membrane bioreactor

A cross-flow membrane was coupled to a chemostat to create an anaerobic membrane bioreactor (MBR) for biological hydrogen production. The reactor was fed glucose (10,000 mg/L) and inoculated with a soil inoculum heat-treated to kill non-spore-forming methanogens. Hydrogen gas was consistently produced at a concentration of 57-60% in the headspace under all conditions. When operated in chemostat mode (no flow through the membrane) at a hydraulic retention time (HRT) of 3.3 h, 90% of the glucose was removed, producing 2200 mg/L of cells and 500 mL/h of biogas. When operated in MBR mode, the solids retention time (SRT) was increased to SRT = 12 h producing a solids concentration in the reactor of 5800 mg/L. This SRT increased the overall glucose utilization (98%), the biogas production rate (640 mL/h), and the conversion efficiency of glucose-to-hydrogen from 22% (no MBR) to 25% (based on a maximum of 4 mol-H(2)/mol-glucose). When the SRT was increased from 5 h to 48 h, glucose utilization (99%) and biomass concentrations (8,800 +/- 600 mg/L) both increased. However, the biogas production decreased (310 +/- 40 mL/h) and the glucose-to-hydrogen conversion efficiency decreased from 37 +/- 4% to 18 +/- 3%. Sustained permeate flows through the membrane were in the range of 57 to 60 L/m(2) h for three different membrane pore sizes (0.3, 0.5, and 0.8 microm). Most (93.7% to 99.3%) of the membrane resistance was due to internal fouling and the reversible cake resistance, and not the membrane itself. Regular backpulsing was essential for maintaining permeate flux through the membrane. Analysis of DNA sequences using ribosomal intergenic spacer analysis indicated bacteria were most closely related to members of Clostridiaceae and Flexibacteraceae, including Clostridium acidisoli CAC237756 (97%), Linmingia china AF481148 (97%), and Cytophaga sp. MDA2507 AF238333 (99%). No PCR amplification of 16s rRNA genes was obtained when archaea-specific primers were used.     

Comparison between a moving bed membrane bioreactor and a conventional membrane bioreactor on membrane fouling

A membrane bioreactor filled with carriers instead of activated sludge named a moving bed membrane bioreactor (MBMBR) was investigated to minimize the effect of suspended solids on membrane fouling. The MBMBR and a conventional membrane bioreactor (CMBR) were operated in parallel for about two months. Unexpectedly, the rate of membrane fouling in MBMBR was about three times of that in CMBR. MBMBR showed a higher cake layer resistance than CMBR due to plenty of filamentous bacteria inhabited in suspended solids in MBMBR. Protein and polysaccharide contents of soluble EPS in MBMBR were obviously larger than those in CMBR. It could be speculated that the overgrowth of filamentous bacteria in MBMBR resulted in severe cake layer and induced a large quantity of EPS, which deteriorated the membrane fouling.     

Enrichment of ANME‐2 dominated anaerobic methanotrophy from cold seep sediment in an external ultrafiltration membrane bioreactor

Anaerobic oxidation of methane (AOM) coupled to sulfate reduction is a microbially mediated unique natural phenomenon with an ecological relevance in the global carbon balance and potential application in biotechnology. This study aimed to enrich an AOM performing microbial community with the main focus on anaerobic methanotrophic archaea (ANME) present in sediments from the Ginsburg mud volcano (Gulf of Cadiz), a known site for AOM, in a membrane bioreactor (MBR) for 726 days at 22 (± 3)°C and at ambient pressure. The MBR was equipped with a cylindrical external ultrafiltration membrane, fed a defined medium containing artificial seawater and operated at a cross flow velocity of 0.02 m/min. Sulfide production with simultaneous sulfate reduction was in equimolar ratio between days 480 and 585 of MBR operation, whereas methane consumption was in oscillating trend. At the end of the MBR operation (day 726), the enriched biomass was incubated with ¹³C labeled methane, ¹³C labeled inorganic carbon was produced and the AOM rate based on ¹³C‐inorganic carbon was 1.2 μmol/(g_dw d). Microbial analysis of the enriched biomass at 400 and 726 days of MBR operation showed that ANME‐2 and Desulfosarcina type sulfate reducing bacteria were enriched in the MBR, which formed closely associated aggregates. The major relevance of this study is the enrichment of an AOM consortium in a MBR system which can assist to explore the ecophysiology of ANME and provides an opportunity to explore the potential application of AOM.     

Optimal operating policy of the ultrafiltration membrane bioreactor for enzymatic hydrolysis of cellulose

The dilution rate of an ultrafiltration membrane bioreactor in the enzymatic hydrolysis of cellulose was optimized using the kinetic model developed by Fan and Lee.(4) The sequence of optimal dilution rates was found to generally consist of an initial period of a minimal value (batch period), a subsequent period of maximum dilution rate, a period of a second batch, and a final period of a singular dilution rate. The effects of operating conditions, such as beta-glucosidase activity, operating time, maximum dilution rate, substrate feeding rate, and enzyme-to-substrate ratio on both the conversion yield and the sequence of optimal dilution rates were investigated. To evaluate the validity of kinetic model employed in this work, enzymatic hydrolysis was carried out using alpha-cellulose as a substrate in the ultrafiltration membrane bioreactor. The experimental data were well consistent with the simulation results. (c) 1993 John Wiley & Sons, Inc.     

Application of a 22L scale membrane bioreactor and cross-flow ultrafiltration to obtain purified chondroitin

Recently, the possibility of producing fructosylated chondroitin from the capsular polysaccharide of Escherichia coli O5:K4:H4, in fed‐batch and microfiltration experiments was assessed on a 2 L bioreactor. In this work, a first scale‐up step was set on a 22 L membrane reactor with modified baffles to insert ad hoc designed microfiltration modules permanently inside the bioreactor vessel. Moreover, the downstream polysaccharide purification process, recently established on the A¨?KTA cross‐flow instrument, was translated to a UNIFLUX‐10, a tangential flow filtration system suitable for prepilot scale. In particular, the microfiltered permeates obtained throughout the fermentation, and the supernatant recovered from the centrifuged broth at the end of the process, were treated as two separate samples in the following ultrafiltration procedure, and the differences in the two streams and how these affected the ultrafiltration/diafiltration process performance were analysed. The total amount of K4 capsular polysaccharide was about 85% in the broth and 15% in the microfiltered permeates. However, the downstream treatment was more efficient when applied to the latter. The major contaminant, the lipopolysaccharide, could easily be separated by a mild hydrolysis that also results in the elimination of the unwanted fructosyl residue, which is linked to the C‐3 of glucuronic acid residues. The tangential ultrafiltration/diafiltration protocols developed in a previous work were effectively scaled‐up, and therefore in this research proof of principle was established for the biotechnological production of chondroitin from the wild‐type strain E. coli O5:K4:H4. The complete downstream procedure yielded about 80% chondroitin with 90% purity. © 2012 American Institute of Chemical Engineers Biotechnol. Prog., 28: 1012–1018, 2012     

氧对膜生物反应器短程硝化的影响

为了研究膜生物反应器的短程硝化性能以及氧对短程硝化的影响,通过对比耗氧率和供氧率,提出了膜生物反应器短程硝化的控制优化建议。在膜生物反应器硝化过程中,DO小于1 mg/L开始出现亚硝氮积累;DO降到0.5 mg/L,出水氨氮浓度与亚硝氮浓度之比接近1∶1;DO调控在0.5-1 mg/L范围内,有利于前置硝化反应器与后续厌氧氨氧化反应器衔接。膜生物反应器中污泥浓度可达20 g/L,耗氧能力可达19.86 mg O2/(L·s),但最大供氧能力仅为0.369 mg O2/(L·s),供氧成为反应器运行的制约瓶颈,"低DO高流量"曝气是继续提高短程硝化效能的控制策略。     

High-level production of D-mannitol with membrane cell-recycle bioreactor

Ten heterofermentative lactic acid bacteria were compared in their ability to produce D-mannitol from D-fructose in a resting state. The best strain, Leuconostoc mesenteroides ATCC-9135, was examined in high cell density membrane cell-recycle cultures. High volumetric mannitol productivity (26.2 g l⁻¹ h⁻¹) and mannitol yield (97 mol%) were achieved. Using the same initial biomass, a stable high-level production of mannitol was maintained for 14 successive bioconversion batches. Applying response surface methodology, the temperature and pH were studied with respect to specific mannitol productivity and yield. Moreover, increasing the initial fructose concentration from 100 to 120 and 140 g l⁻¹ resulted in decreased productivities due to both substrate and end-product inhibition of the key enzyme, mannitol dehydrogenase (MDH). Nitrogen gas flushing of the bioconversion media was unnecessary, since it did not change the essential process parameters. Journal of Industrial Microbiology & Biotechnology (2002) 29, 44–49 doi:10.1038/sj.jim.7000262 Received 12 November 2001/ Accepted in revised form 30 March 2002     

Operational patterns affecting lactic acid production in ultrafiltration cell recycle bioreactor

Lactic acid production with cell recycling on an ultrafiltration tubular membrane reactor was studied; higher lactic acid concentrations as well as productivities were obtained under long-term fermentations compared with other high cell density systems. Different operational conditions, namely dilution rates and start-up modes, were assessed. Performances were very different at the three different dilution rates tested (D = 0.20 h(-1), D = 0.40 h(-1), or D = 0.58 h(-1)). The different behaviours are discussed and factors responsible for them are presented. The best way to operate for lactic acid production is chosen, the dilution rate of D = 0.40 h(-1) being the one providing the best overall performance. On the other hand, results show that of the two start-up modes tested, continuous start (membrane open) permits higher permeabilities throughout the operational runs than batch start (membrane closed). Operational stability was found to be directly associated with membranes that work at "steady state," the membrane permeability being kept around 15 L/m(2) h. Optimized cell bleed can improve time of operation if such membrane permeability can be maintained for a longer time. A comparison of results with those obtained in other lactic acid production systems is presented; such comparison shows that this tubular ultrafiltration membrane cell recycle reactor presents three important advantages: (1) concomitant lactic acid concentrations and productivities; (2) long periods of operation at reasonable permeabilities; and (3) good mechanical stability permitting the use of steam sterilization. (c) 1995 John Wiley & Sons, Inc.     

EPS and SMP dynamics at different heights of a submerged anaerobic membrane bioreactor (SAMBR)

Membrane bioreactors (MBR) technology for wastewater offers many advantages over conventional technologies such as high effluent quality, less footprint and others. The main disadvantage of membrane bioreactors (MBR) is related to membrane fouling, which is mainly caused by extracellular polymeric substance (EPS) and soluble microbial products (SMP). This research studied EPS and SMP dynamics at different heights of a submerged anaerobic membrane bioreactor (SAMBR). The SAMBR was operated under two organic loading rates (OLR) (0.79 and 1.56 kg/m³ d) and was fed with synthetic wastewater with glucose as the carbon source. The results showed percentages of chemical oxygen demand (COD) removal above 95% and the highest COD removal rates were observed at the bottom of the reactor (>83%) for both OLR. The EPS showed a stratification with highest quantities in the supernatant. For the SMP the highest concentration was in the bottom of SAMBR where utilization predominated associated products whereas in the SAMBR supernatant predominated biomass associated products. The OLR change led to a significant increase in SMP accumulation but not in EPS. These facts showed that EPS and SMP dynamic in the SAMBR seemed to be mainly influenced by biological activity, total suspended solids concentration and substrate composition.     

Membrane fouling in a membrane bioreactor (MBR): sludge cake formation and fouling characteristics

A submerged membrane bioreactor (MBR) with a working volume of 1.4 L and a hollow fiber microfiltration membrane was used to treat a contaminated raw water supply at a short hydraulic retention time (HRT) of approximately 1 h. Filtration flux tests were conducted regularly on the membrane to determine various fouling resistances, and confocal laser scanning microscopy (CLSM) and scanning electron microscopy (SEM) were employed to characterize the biofouling development and sludge cake formation on the membrane. The experimental results demonstrate that the MBR is highly effective in drinking water treatment for the removal of organic pollutants, ammonia, and UV absorbance. During the MBR operation, the fouling materials were not uniformly distributed on the entire surface of all of the membrane fibers. The membrane was covered partially by a static sludge cake that could not be removed by the shear force of aeration, and partially by a thin sludge film that was frequently washed away by aeration turbulence. The filtration resistance coefficients were 308.4 x 10(11) m(-1) on average for the sludge cake, 32.5 x 10(11) m(-1) on average for the dynamic sludge film, and increased from 10.5 x 10(11) to 59.7 x 10(11) m(-1) for the membrane pore fouling after 10 weeks of MBR operation at a filtration flux of 0.5 m3/m2 x d. Polysaccharides and other biopolymers were found to accumulate on the membrane, and hence decreased membrane permeability. More important, the adsorption of biopolymers on the membrane modified its surface property and led to easier biomass attachment and tighter sludge cake deposition, which resulted in a progressive sludge cake growth and serious membrane fouling. The sludge cake coverage on the membrane can be minimized by the separation, with adequate space, of the membrane filters, to which sufficient aeration turbulence can then be applied.     

Design and evaluation of a continuous semipartition bioreactor for in situ liquid‐liquid extractive fermentation

Extractive fermentation (or in situ product removal (ISPR)) is an operational method used to combat product inhibition in fermentations. To achieve ISPR, different separation techniques, modes of operation and physical reactor configurations have been proposed. However, the relative paucity of industrial application necessitates continued investigation into reactor systems. This article outlines a bioreactor designed to facilitate in situ product extraction and recovery, through adapting the reaction volume to include a settler and solvent extraction and recycle section. This semipartition bioreactor is proposed as a new mode of operation for continuous liquid‐liquid extractive fermentation. The design is demonstrated as a modified bench‐top fermentation vessel, initially analysed in terms of fluid dynamic studies, in a model two‐liquid phase system. A continuous abiotic simulation of lactic acid (LA) fermentation is then demonstrated. The results show that mixing in the main reaction vessel is unaffected by the inserted settling zone, and that the size of the settling tube effects the maximum volumetric removal rate. In these tests the largest settling tube gave a potential continuous volumetric removal rate of 7.63 ml/min; sufficiently large to allow for continuous product extraction even in a highly productive fermentation. To demonstrate the applicability of the developed reactor, an abiotic simulation of a LA fermentation was performed. LA was added to reactor continuously at a rate of 33ml/h, while continuous in situ extraction removed the LA using 15% trioctylamine in oleyl alcohol. The reactor showed stable LA concentration of 1 g/L, with the balance of the LA successfully extracted and recovered using back extraction. This study demonstrates a potentially useful physical configuration for continuous in situ extraction.     

Biodegradation of BTEX vapors in a silicone membrane bioreactor system

The biotreatment of complex mixtures of volatile organic compounds (VOCs) such as benzene, toluene, ethylbenzene, and xylene isomers (BTEX) has been investigated by many workers. However, the majority of the work has dealt with the treatment of aqueous or soil phase contamination. The biological treatment of gas and vapor phase sources of VOC wastes has recently received attention with increased usage of biofilters and bioscrubbers. Although these systems are relatively inexpensive, performance problems associated with biomass plugging, gas channeling, and support media acidification have limited their adoption. In this report we describe the development and evaluation of an alternative biotreatment system that allows rapid diffusion of both BTEX and oxygen through a silicone membrane to an active biofilm. The bioreactor system has a rapid liquid recycle, which facilitates nutrient medium mixing over the biofilm and allows for removal of sloughing cell mass. The system removed BTEX at rates up to 30 μg h⁻¹ cm⁻² of membrane area. BTEX removal efficiencies ranged from 75% to 99% depending on the BTEX concentration and vapor flowrate. Consequently, the system can be used for continuous removal and destruction of BTEX and other potential target VOCs in vapor phase streams. Journal of Industrial Microbiology & Biotechnology (2001) 26, 316–325. Received 14 August 2000/ Accepted in revised form 28 February 2001     

Optimization and scale up of itaconic acid production from potato starch waste in stirred tank bioreactor

Present study used Aspergillus terreus strain C1 isolated from mangrove soil for itaconic acid (IA) production from potato starch waste. Fermentation parameters were optimized by classical one factor approach and statistical experimental designs, such as Plackett-Burman and response surface designs. Anionic deionization of potato waste was found to be a very effective, economic, and easy way of improving IA production. The increase in IA production by deionization was found to correlate with removal of phosphate. In our knowledge, this is the first report on application of deionization of potato waste to enhance IA production. Other parameters like inoculum development conditions, pH, presence of peptone and certain salts in the medium also significantly affected IA production. IA production by strain C1 increased 143-fold during optimization when compared with the starting condition. The optimized IA level (35.75 g/L) was very close to the maximum production predicted by RSM (38.88 g/L). Bench scale production of IA was further optimized in 3-L stirred tank reactor by varying parameters like agitation and aeration rate. The maximum IA production of 29.69 g/L was obtained under the agitation speed of 200 rpm and aeration rate of 0.25 vvm. To the best of our knowledge, it is the first report on IA production from potato starch waste at bioreactor level. © 2019 American Institute of Chemical Engineers Biotechnol. Prog., 35: e2774, 2019.     

Removal of organic pollutants and analysis of MLSS–COD removal relationship at different HRTs in a submerged membrane bioreactor

In order to investigate the influence of hydraulic retention time (HRT) on organic pollutant removal in a submerged membrane bioreactor (SMBR), a laboratory-scale experiment was conducted using domestic sewage as influent. The dissolved oxygen (DO) concentration was controlled at 2.0– during the experimental period. The experiments demonstrated that when HRT was 3, 2 and 1 h, the reduction of chemical oxygen demand (COD) was 89.3–97.2, 88.5–97.3 and 80–91.1%, and the effluent COD was 38.9–11.2, 41.6–10.8 and 63.4–, respectively. It is suggested that an HRT of 1 h could meet the normal standard of discharged domestic sewage, and an HRT of 2 h could meet that of water reclamation. In addition, we use mathematical software MATLAB to analyse the relation of mixed liquor suspended solids (MLSS) and COD removal. The results showed that the optimum MLSS concentration should be maintained at around in the SMBR. The results also showed that the COD removal was related to HRT (τ), influent concentration (S₀) and sludge loading rate for COD removal (N_S). Moreover, the high COD removal could be achieved through adjusting τ, S₀ and N_S.