Ex situ product recovery for enhanced butanol production by Clostridium beijerinckii.

In situ butanol recovery fermentation has been intensively studied as an effective alternative to conventional butanol production, which is limited due to the cellular toxicity of butanol. However, the low biocompatibility of adsorbents often leads to failure of in situ recovery fermentations. In this study, Clostridium beijerinckii NCIMB 8052 was cultured in flasks without shaking and in situ recovery fermentation was performed by using an adsorbent L493. The amounts of acetone, butanol, and ethanol (ABE) increased by 34.4 % in the presence of the adsorbent. In contrast, cell growth and production of organic acids and ABE were retarded in the 7-L batch fermentations with in situ butanol recovery. Cell damage occurred in the fermentor upon agitation in the presence of the adsorbent, unlike in static flask cultures with in situ recovery. Ex situ recovery fermentation using circulation of fermentation broth after mid-exponential phase of cell growth was developed to avoid adsorbent-cell incompatibility. No apparent cell damage was observed and 25.7 g/L of ABE was produced from 86.2 g/L glucose in the fed-batch mode using 7 L fermentors. Thus, ex situ recovery fermentation with C. beijerinckii is effective for enhancing butanol fermentation.

Volatile fatty acids derived from waste organics provide an economical carbon source for microbial lipids/biodiesel production.

Volatile fatty acids (VFAs) derived from organic waste, were used as a low cost carbon source for high bioreactor productivity and titer. A multi-stage continuous high cell density culture (MSC-HCDC) process was employed for economic assessment of microbial lipids for biodiesel production. In a simulation study we used a lipid yield of 0.3 g/g-VFAs, cell mass yield of 0.5 g/g-glucose or wood hydrolyzates, and employed process variables including lipid contents from 10-90% of cell mass, bioreactor productivity of 0.5-48 g/L/h, and plant capacity of 20000-1000000 metric ton (MT)/year. A production cost of USD 1.048/kg-lipid was predicted with raw material costs of USD 0.2/kg for wood hydrolyzates and USD 0.15/kg for VFAs; 9 g/L/h bioreactor productivity; 100, 000 MT/year production capacity; and 75% lipids content. The variables having the highest impact on microbial lipid production costs were the cost of VFAs and lipid yield, followed by lipid content, fermenter cost, and lipid productivity. The cost of raw materials accounted for 66.25% of total operating costs. This study shows that biodiesel from microbial lipids has the potential to become competitive with diesels from other sources.

Multi-stage continuous high cell density culture systems: a review.

A multi-stage continuous high cell density culture (MSC-HCDC) system makes it possible to achieve high productivity together with high product titer of many bioproducts. For long-term continuous operation of MSC-HCDC systems, the cell retention time and hydraulic retention time must be decoupled and strains (bacteria, yeast, plant, and animal cells) must be stable. MSC-HCDC systems are suitable for low-value high-volume extracellular products such as fuel ethanol, lactic acid or volatile fatty acids, and high-value products such as monoclonal antibodies as well as intracellular products such as polyhydroxybutyric acid (PHB), microbial lipids or a number of therapeutics. Better understanding of the fermentation kinetics of a specific product and reliable high-density culture methods for the product-generating microorganisms will facilitate timely industrialization of MSC-HCDC systems for products that are currently obtained in fed-batch bioreactors.

The effect of volatile fatty acids as a sole carbon source on lipid accumulation by Cryptococcus albidus for biodiesel production.

The use of volatile fatty acids (VFAs) for microbial lipid accumulation was investigated in flask cultures of Cryptococcus albidus. The optimum culture temperature and pH were 25°C and pH 6.0, respectively, and the highest lipid content (27.8%) was obtained with ammonia chloride as a nitrogen source. The lipid yield coefficient on VFAs was 0.167 g/g of C. albidus with a VFAs (acetic, propionic, butyric acids) ratio of 8:1:1, which was in good agreement with a theoretically predicted lipid yield coefficient of the VFAs as a carbon source. The major fatty acids of the lipids accumulated by C. albidus were similar to those of soybean oil and jatropha oil. A preliminary cost analysis shows that VFAs-based biodiesel production is competitive with current palm and soybean based biodiesels. Further process development for lower aeration cost and higher lipid yield will make this process more economical.

Performance of microbial fuel cell with volatile fatty acids from food wastes.

Food wastes were used as feedstock for the direct production of electricity in a microbial fuel cell (MFC). MFC operations with volatile fatty acids (VFA) produced 533 mV with a maximum power density of 240 mW/m(2). Short-chain VFAs, such as acetate, were degraded more rapidly and thus supported higher power generation than longer chain ones. In general, the co-existence of other, different VFAs slowed the removal of each VFA, which indicated that anodic microbes were competing for different substrates. 16S rRNA gene analysis using PCR-DGGE indicated that the MFC operation with VFAs had enriched unique microbial species.

Multi-stage high cell continuous fermentation for high productivity and titer.

We carried out the first simulation on multi-stage continuous high cell density culture (MSC-HCDC) to show that the MSC-HCDC can achieve batch/fed-batch product titer with much higher productivity to the fed-batch productivity using published fermentation kinetics of lactic acid, penicillin and ethanol. The system under consideration consists of n-serially connected continuous stirred-tank reactors (CSTRs) with either hollow fiber cell recycling or cell immobilization for high cell-density culture. In each CSTR substrate supply and product removal are possible. Penicillin production is severely limited by glucose metabolite repression that requires multi-CSTR glucose feeding. An 8-stage C-HCDC lactic acid fermentation resulted in 212.9 g/L of titer and 10.6 g/L/h of productivity, corresponding to 101 and 429% of the comparable lactic acid fed-batch, respectively. The penicillin production model predicted 149% (0.085 g/L/h) of productivity in 8-stage C-HCDC with 40 g/L of cell density and 289% of productivity (0.165 g/L/h) in 7-stage C-HCDC with 60 g/L of cell density compared with referring batch cultivations. A 2-stage C-HCDC ethanol experimental run showed 107% titer and 257% productivity of the batch system having 88.8 g/L of titer and 3.7 g/L/h of productivity. MSC-HCDC can give much higher productivity than batch/fed-batch system, and yield a several percentage higher titer as well. The productivity ratio of MSC-HCDC over batch/fed-batch system is given as a multiplication of system dilution rate of MSC-HCDC and cycle time of batch/fed-batch system. We suggest MSC-HCDC as a new production platform for various fermentation products including monoclonal antibody.

Anaerobic organic acid production of food waste in once-a-day feeding and drawing-off bioreactor.

Acidogenesis of food waste was studied in a 2-L reactor with semi-continuous mode operation (once-a-day feeding and draw-off) for maximum 65 days to examine optimal volatile acid compositions for biological nitrogen removal (BNR) and enhanced biological phosphorus removal (ENPR). Various operational parameters of hydraulic retention time (HRT), organic loading rate (ORL), pH and temperature were investigated for soluble chemical oxygen demand (SCOD), volatile fatty acid composition, nitrogen and phosphate. The yields (gTVFA/g VS) and the volumetric productivity (gTVFA/d L) increased with HRT from 0.26-0.32, 1.25-1.50 (at 4 days) to 0.36-0.39, 1.71-1.83 (at 12 days). However, the acetate fraction (%) decreased with HRT from 35.7-37.5 at 4 days to 23.5-25 at 12 days. The yields decreased with increase of organic loading from 0.34-0.37 at 5 g/L d to 0.29-0.30 at 13 g/L d and the productivity increased from 1.63-1.65 to 3.61-3.75. The yield and productivity were highest at 35 degrees C among 25, 35 and 45 degrees C. The yield and productivity at pH 5.5 and 6.0 were best and very similar to each other. The condition of 35 degrees C, pH 6.0, HRT 8 days, ORL 9 g/L d resulted in TVFA, SCOD, acetate and butyrate of 25, 39.5, 12 and 5.25 g/L, respectively.