Kinetic and growth parameters of Arthrospira (Spirulina) platensis cultivated in tubular photobioreactor under different cell circulation systems

Arthrospira platensis was cultivated in tubular photobioreactor in order to evaluate growth and biomass production at variable photosynthetic photon flux density (PPFD = 60, 120, and 240 μmol photons m(-2)s(-1)) and employing three different systems for cell circulation, specifically an airlift, a motor-driven pumping and a pressurized system. The influence of these two independents variables on the maximum cell concentration (X(m)), cell productivity (P(x)), nitrogen-to-cell conversion factor (Y(X/N) ), photosynthetic efficiency (PE), and biomass composition (total lipids and proteins), taken as responses, was evaluated by analysis of variance. The statistical analysis revealed that the best combination of responses' mean values (X(m) = 4,055 mg L(-1), P(x) = 406 mg L(-1)day(-1), Y(X/N) = 5.07 mg mg(-1), total lipids = 8.94%, total proteins = 30.3%, PE = 2.04%) was obtained at PPFD = 120 μmol photons m(-2)s(-1); therefore, this light intensity should be considered as the most well-suited for A. platensis cultivation in this photobioreactor configuration. The airlift system did not exert any significant positive statistical influence on the responses, which suggests that this traditional cell circulation system could successfully be substituted by the others tested in this work.     

Repeated-batch cultures of Baby Hamster Kidney cell aggregates in stirred vessels

Natural aggregates of Baby Hamster Kidney cells were grown in stirred vessels operated as repeated-batch cultures during more than 600 hours. Different protocols were applied to passaging different fractions of the initial culture: single cells, large size distributed aggregates and large aggregates. When single cells or aggregates with the same size distribution found in culture are used as inoculum, it is possible to maintain semi-continuous cultures during more than 600 hours while keeping cell growth and viability. These results suggest that aggregate culture in large scale might be feasible, since a small scale culture can easily be used as inoculum for larger vessels without noticeable modification of the aggregate chacteristics. However, when only the large aggregates are used as inoculum, it was shown that much lower cell concentrations are obtained, cell viability in aggregates dropping to less than 60%. Under this selection procedure, aggregates maintain a constant size, larger than under batch experiments, up to approximately 400 hours; after this time, aggregate size increases to almost twice the size expected from batch cultures.     

一种用于微藻培养的新型板式光生物反应器

微藻的闪光效应可以大幅提高微藻的光效率,提高微藻产量。通过在传统的板式光生物反应器中加入斜挡板以增强微藻的闪光效应。以小球藻为模型藻种,考察了新型板式光生物反应器内不同光强和不同进口流速对小球藻生长速率和光效率的影响。结果表明,当进口流速为0.16 m/s时,随着光强的提高,小球藻的细胞浓度逐渐增加,光效率逐渐降低;在500μmol/(m2·s)的光强条件下,小球藻细胞浓度和光效率均随着进口流速的提高而增加。新型板式光生物反应器内小球藻的细胞浓度比传统板式光生物反应器提高了39.23%,表明在传统板式光生物反应器内加入斜挡板可有效增强微藻的闪光效应。     

Lipase-catalyzed selective synthesis of monolauroyl maltose using continuous stirred tank reactor

Monolauroyl maltose was synthesized by an immobilized lipase that catalyzed condensation of maltose and lauric acid in acetone using a batch reactor or a continuous stirred tank reactor. Mono- and di-lauroyl maltoses were identified by FT-IR, ¹H NMR, ¹³C NMR and MS. Monolauroyl maltose was selectively synthesized in a continuous stirred tank reactor and no diester was detected. The highest concentration of monolauroyl maltose at 28 mmol/l was obtained in 250 ml acetone when maltose was added at 4 g/d and the molar ratio of lauric acid to maltose was fixed at 4:1 at a flow rate of 0.15 ml/min for both influx and effluent without supplement of fresh molecular sieve.     

Production of mouse monoclonal antibodies using a continuous cell culture fermenter and protein G affinity chromatography

The production of anti--fetoprotein monoclonal antibodies for diagnostic use was carried out in a stirred tank fermenter equipped with a double membrane stirrer for bubble free aeration and continuous medium perfusion. A serum-free medium supplemented with 4 mM L-glutamine and 2.0 g/l glucose with a protein content of only 780 g/ml was used for the production process. The harvested antibodies were concentrated 50-fold using a tangential ultrafiltration system and were then purified in a one step purification process by protein G affinity chromatography. The purity of the final product (90%) was controlled by SDS-polyacrylamide gel electrophoresis, gel exclusion chromatography and isoelectric focussing. For further quality controls of the product the immunoglobulin subclass and the isoelectric point were determined and the specificity of the purified mAb was tested by RIA using¹²⁵I labelled -fetoprotein.1.87 g of purified monoclonal antibodies were produced (90% purity) within 2 weeks. It was found that the use of this type of stirred tank fermenter combined with a one step purification process using protein G affinity chromatography represents a suitable method for the fast production of medium scale quantities (500 mg–5 g) of monoclonal antibodies for diagnostic use.Abbreviations AFP -Fetoprotein - BSA bovine serum albumine - FCS Fetal calf serum - HRP horseradish peroxidase - OPD o-phenylenediamine dihydrochloride - I.P. isoelectric point - IEF isoelectric focussing - PBS Phosphate buffered saline     

Optimization of L-malic acid production by Aspergillus flavus in a stirred fermentor

Effects of various nutritional and environmental factors on the accumulation of organic acids (mainly L-malic acid) by the filamentous fungus Aspergillus flavus were studied in a 16-L stirred fermentor. Improvement of the molar yield (moles acid produced per moles glucose consumed) of L-malic acid was obtained mainly by increasing the agitation rate (to 350 rpm) and the Fe(z+) ion concentration (to 12 mg/L) and by lowering the nitrogen (to 271 mg/L) and phosphate concentrations (to 1.5 mM) in the medium. These changes resulted in molar yields for L-malic acid and total C(4) acids (L-malic, succinic, and fumaric acids) of 128 and 155%, respectively. The high molar yields obtained (above 100%) are additional evidence for the operation of part of the reductive branch of the tricarboxylic acid cycle in L-malic acid accumulation by A. flavus. The fermentation conditions developed using the above mentioned factors and 9% CaCO(3) in the medium resulted in a high concentration (113 g/L L-malic acid from 120 g/L glucose utilized) and a high overall productivity (0.59 g/L h) of L-malic acid. These changes in acid accumulation coincide with increases in the activities of NAD(+)-malate dehydrogenase, fumarase, and citrate synthase.     

Production cost of a real microalgae production plant and strategies to reduce it

The cost analysis of a real facility for the production of high value microalgae biomass is presented. The facility is based on ten 3 m³ tubular photobioreactors operated in continuous mode for 2 years, data of Scenedesmus almeriensis productivity but also of nutrients and power consumption from this facility being used. The yield of the facility was close to maximum expected for the location of Almería, the annual production capacity being 3.8 t/year (90 t/ha·year) and the photosynthetic efficiency being 3.6%. The production cost was 69 €/kg. Economic analysis shows that labor and depreciation are the major factors contributing to this cost. Simplification of the technology and scale-up to a production capacity of 200 t/year allows to reduce the production cost up to 12.6 €/kg. Moreover, to reduce the microalgae production cost to approaches the energy or commodities markets it is necessary to reduce the photobioreactor cost (by simplifying its design or materials used), use waste water and flue gases, and reduce the power consumption and labor required for the production step. It can be concluded that although it has been reported that production of biofuels from microalgae is relatively close to being economically feasible, data here reported demonstrated that to achieve it by using the current production technologies, it is necessary to substantially reduce their costs and to operate them near their optimum values.     

Alkaloid production by plant cell cultures of Holarrhena antidysenterica: II. Effect of precursor feeding and cultivation in stirred tank bioreactor

Precursor feeding strategy for increasing the yield of conessine, a steroidal alkaloid of Holarrhena antidysenterica, was established in cell suspension culture. A total of 50 mg/L added cholesterol was converted into 43 mg/L of alkaloid, 90% of which constituted the conessine. By applying the precursor feeding policy to the cell suspension culture in modified Murashige and Skoog (MS) medium, a total of 143 mg/L of alkaloid was produced in 8 days. In this way the alkaloid content of the cells was increased more than six times compared to that obtained in the standard MS medium. The steps leading to biotransformation of cholesterol into alkaloids were unaffected by phosphate. The shake flask data were successfully transferred to a bench scale 6-L stirred tank bioreactor in which the specific biosynthetic rate of alkaloid production was 110 mg/100 g dry cell weight per day, about 160 times higher than that of whole plant.     

Innovative development of membrane sparger for carbon dioxide supply in microalgae cultures

The present study was aimed to develop a membrane sparger (MS) integrated into a tubular photobioreactor to promote the increase of the carbon dioxide (CO₂) fixation by Spirulina sp. LEB 18 cultures. The use of MS for the CO₂ supply in Spirulina cultures resulted not only in the increase of DIC concentrations but also in the highest accumulated DIC concentration in the liquid medium (127.4 mg L⁻¹ d⁻¹). The highest values of biomass concentration (1.98 g L⁻¹), biomass productivity (131.8 mg L⁻¹ d⁻¹), carbon in biomass (47.9% w w⁻¹), CO₂ fixation rate (231.6 mg L⁻¹ d⁻¹), and CO₂ use efficiency (80.5% w w⁻¹) by Spirulina were verified with MS, compared to the culture with conventional sparger for CO₂ supply. Spirulina biomass in both culture conditions had high protein contents varying from 64.9 to 69% (w w⁻¹). MS can be considered an innovative system for the supply of carbon for the microalgae cultivation and biomass production. Moreover, the use of membrane system might contribute to increased process efficiency with a reduced cost of biomass production.