Production of α‐amylase under solid‐state fermentation utilizing coffee waste

Coffee industry substrates such as coffee pulp, coffee cherry husk, silver skin, spent coffee and mixtures of these coffee wastes (MC) were evaluated for their efficacy as sole carbon source for the synthesis of α‐amylase in solid‐state fermentation (SSF) using a fungal strain of Neurospora crassa CFR 308. For SSF with coffee pulp and with MC, α‐amylase activity of 3908 U g^?1 ds (units per gram of dry substrate) and 3870 U g^?1 ds, respectively, was observed. Parameters such as moisture (60%), pH (4.6), temperature (28 °C), particle size (1.0 mm), inoculum size (10⁷ spores g^?1 ds), and fermentation time (5 days) were optimized for enzyme synthesis, wherein 4981 and 4324 U g^?1 U g^?1 ds of α‐amylase activity was obtained in SSF with coffee pulp and MC, respectively. The enzyme production was further improved when the substrates were subjected to pre‐treatment by steaming. Accordingly, maximum α‐amylase activity of 7084 U g^?1 ds and 6342 U g^?1 ds was obtained with steam‐pretreated coffee pulp and MC, respectively, demonstrating them to be excellent sole carbon sources for synthesis of α‐amylase production. Copyright © 2009 Society of Chemical Industry     

Continuous alcohol production from whey permeate using immobilised cell reactor systems

The present paper reports the performance of a bioreactor packed with alginate-entrapped Kluyveromyces marxianus NCYC179 for continuous fermentation of whey permeate to ethanol. A maximum ethanol productivity as 28.21 gl^?1 h^?1 was attained at D=0.42h^?1 and 75% lactose consumption (substrate feed rate in the inflowing medium was 200 g lactose I^?1). However, the higher dilution rates (0.6-1.Oh^?1) resulted in poor productivities and higher substrate washout in the effluent samples. The maximum specific ethanol production (qpi) and maximum specific lactose uptake (qsi) of the immobilised Kluyveromyces marxianus NCYC179 was found to be 3.88g ethanol/g immobilised cell/hx10^?2 and 8.75g lactose consumed/g immobilised cell/hx10^?2 respectively. A bead size of 2.5 mm in diameter and activation period of 24h of alginate beads in lactose solution (10%) prior to their packing in column reactor were found to support the efficient working of the bioreactor. The immobilised cell bioreactor system was operated continuously at a constant dilution rate of 0.15h^?1 and 10% lactose for 562 h without any significant change in the efficiency (varied from 84 to 88% of theoretical) and viability of the entrapped yeast cells (dropped from 84 to 81%).     

Novel two‐in‐one bioreactor greatly improves lactic acid production from xylose by Lactobacillus pentosus

BACKGROUND: Simultaneous xylose isomerization and fermentation was investigated to improve the lactic acid production from xylose by Lactobacillus pentosus in a novel two‐in‐one bioreactor constructed by packing the immobilized xylose isomerase (65 g) in a fixed bed reactor (diameter 56 mm × 66 mm, packing volume 154 mL) with a permeable wall, which was installed inside a conventional fermenter (2 L) and rotated along the axis together with the mechanical stirrer of the fermenter. RESULTS: Xylose (20 g L^?1) was completely consumed within 24 h in the novel bioreactor, compared with 72 h needed for the control without packed enzyme. The maximum cell density (17.5 g L^?1) in the novel bioreactor was twice that in the control and the lactic acid productivity (0.58 g L^?1 h^?1) was 3.8 times higher. Repeated use of the immobilized enzyme showed that the lactic acid productivity and yield obviously dropped after the first batch fermentation but maintained almost unchanged afterwards. CONCLUSION: Simultaneous xylose isomerization and fermentation significantly improved lactic acid production from xylose by Lactobacillus pentosus. The novel bioreactor made it easier to recycle and reuse the immobilized enzyme. © 2012 Society of Chemical Industry     

Effects of culture temperature on the production of bioactive polysaccharides by Agaricus blazei in batch cultures

Effects of culture temperature ranging from 20 to 34 °C on cell growth, polysaccharide biosynthesis and the bioactivity of polysaccharides of Agaricus blazei were evaluated via eight batch cultures in steps of 2 °C in a stirred tank bioreactor. Results indicated that the optimal temperature for the biomass was 28 °C with a cell yield of 780 mg g^?1, while that for polysaccharide formation was 30 °C with a product yield of 230 mg g^?1. Both the β‐glucan content and average molecular weight of the polysaccharides from different temperature‐controlled cultures were closely correlated with their tumour necrosis factor‐α (TNF‐α) release capability on macrophage cells. The polysaccharides from the low temperature range (20–24 °C) not only had higher relative content of β‐glucan and average molecular weight but also exhibited higher bioactivity compared with those from the high temperature range (30–34 °C). The optimal temperature for the production of bioactive polysaccharides of A. blazei was 24 °C, at which their relatively high molecular weight, β‐glucan content and TNF‐α release capability on macrophage cells were 1650 kDa, 188 mg g^?1 and 1560 pg per 5 × 10⁴ cells respectively. Copyright © 2007 Society of Chemical Industry     

Optimization of inulinase production by solid‐state fermentation in a packed‐bed bioreactor

BACKGROUND: This work is focused on inulinase production by solid‐sate fermentation (SSF) using sugarcane bagasse, corn steep liquor (CSL), pre‐treated cane molasses, and soybean bran as substrates in a 3‐kg (dry basis) packed‐bed bioreactor. SSF was carried out by the yeast Kluyveromyces marxianus NRRL Y‐7571 and response surface methodology was used to optimize the temperature, air flow rate and initial mass of cells. RESULTS: The optimum inulinase activity (436.7 ± 36.3 U g^?1 dry substrate) was obtained at 24 h at an inlet air temperature of 30 °C, air flow rate 2.2 m³ h^?1 and 22 g of cells for fermentation. Inulinase productivity at these conditions was 18.2 U gds^?1 h^?1. Kinetic evaluation at the optimized conditions showed that the maximum inulinase production was verified at 24 h of fermentation. The carbon dioxide and the metabolic heat generation are directly associated with the consumption of total reducing sugars present in the medium. CONCLUSION: The high productivity achieved in this work shows the technical viability of inulinase production by SSF in a packed‐bed bioreactor. Copyright © 2009 Society of Chemical Industry     

Improved cultivation conditions for polysaccharide production by H. influenzae type b

Haemophilus influenzae b (Hib), an encapsulated Gram‐negative cocco‐bacillus, is one of the most common agents of meningitis worldwide. The capsular polysaccharide conjugated to a carrier protein is the antigen of the vaccine against Hib. An optimized cultivation process that could lead to an increase in the polysaccharide production would be of great interest for mass vaccination programs. The aim of this work was to evaluate different culture conditions in attempt to improve the capsular polysaccharide yield. Hib was cultivated in a bioreactor with modified soy‐peptone and yeast‐extract (MP) medium and optimal hemin and nicotinamide adenine dinucleotide (NAD) concentration in the culture medium was established at 30 mg L^?1 and 15 mg L^?1, respectively. The batch experiments were carried out as follows: (a) overlay aeration without pH control; (b) air‐sparged with dissolved oxygen tension (DOT) controlled at 10 and 30% air saturation, with and without pH control. The cultures with air‐sparged aeration, without pH control, showed values for the specific production (SP_p/x) of 180–190 mg PRP g^?1 dry cell weight (DCW) and overall polysaccharide productivity of 22–29 mg L^?1 h^?1, accounting for an increase of ca 47% over the polysaccharide production with overlay aeration. Batch cultivations with air sparged aeration led to an improvement in the poly(ribosylribitol phosphate) (PRP) production for both conditions (DOT at 10 and 30% air saturation) investigated upon pH control, achieving up to 980 PRP mg L^?1. The SP_p/x and overall polysaccharide productivity were 280–300 mg PRP g^?1 DCW and 45–41 mg L^?1h^?1, respectively. The best production of capsular polysaccharide was obtained in the modified MP‐medium, with 30 mg L^?1 hemin and 15 mg L^?1 NAD, upon sparged aeration and pH control. Copyright © 2005 Society of Chemical Industry     

Penicillium commune spore production in solid‐state fermentation of coffee pulp at laboratory scale and in a helical ribbons rotating reactor

Penicillium commune was grown on coffee pulp (CP) by solid‐state fermentation (SSF). The effects of the duration of CP thermal treatment and the effects of incubation temperature on spore production yield were studied at laboratory scale. The effect of mixing during fermentation was assayed at pilot plant scale in a 70 L stainless steel non‐aseptic reactor equipped with helical ribbons for mixing solids. For thermal treatments of CP at 121 °C for 10, 20, 30 and 40 min, no significant difference in spore production yield was observed. Maximum sporulation yield was found at 25 °C; when the incubation temperature was higher than 30 °C, the sporulation yield decreased significantly. A spore production yield of 3.7 × 10⁹ spores g^?1 dry CP was obtained when continuous mixing (0.25 rpm) was used at pilot plant scale; however, a decrease in spore yield (1.4 × 10⁹ spores g^?1 dry CP) was observed under static conditions. Spore production was not affected when a scale factor between 79 and 105 was assayed from laboratory to pilot plant; at this level, the productivity obtained was 3.1 × 10⁷ spores g^?1 dry CP h^?1. This value is similar to that found in other reports using natural substrates but working at a smaller scale. Copyright © 2006 Society of Chemical Industry     

Ammonium fumarate production by free or immobilised Rhizopus arrhizus in bench‐ and laboratory‐scale bioreactors

Ammonium fumarate production from glucose‐based media by Rhizopus arrhizus NRRL 1526 with mycelial growth controlled by phosphorus limitation exhibited mixed‐growth‐associated product formation kinetics, with growth‐associated production related to secondary mycelial growth only. The contribution of the primary mycelial growth phase was minimised by resorting to prolonged batch production using free mycelia under intermittent glucose feeding or repeated batch production using immobilised mycelia. The metabolic activity of free or immobilised mycelia was limited by fumarate accumulation or by oxygen diffusion phenomena, respectively. For batch cultures in a 15 dm³ stirred bioreactor the peripheral impeller speed (v_I) was increased from 1.88 to 3.3 m s^?1, and the fumarate yield coefficient on glucose increased from 0.25 ± 0.01 to 0.42 ± 0.02 g g^?1, while the malate yield coefficient on fumarate (Y_M/F) reduced from 0.46 ± 0.01 to 0.14 ± 0.01 g g^?1. With a net increase in the fumarate‐to‐malate ratio from 2 to 6.5, a v_I value of 3.3 m s^?1 gave the best fermentation performance and provided a basis for further scale‐up studies. © 2002 Society of Chemical Industry     

Fermentation of cheese whey powder solution to ethanol in a packed‐column bioreactor: effects of feed sugar concentration

BACKGROUND: Cheese whey powder (CWP) is a concentrated source of lactose and other essential nutrients for ethanol fermentation. CWP solution containing different concentrations of total sugar was fermented to ethanol in an up‐flow packed‐column bioreactor (PCBR) at a constant hydraulic residence time (HRT) of 50 h. Total sugar concentration in the feed was varied between 50 and 200 g L^?1 and a pure culture of Kluyveromyces marxianus was used for ethanol fermentation of lactose. Variations of ethanol and sugar concentrations with the height of the column and with the feed sugar concentration were determined. RESULTS: Ethanol concentration increased and total sugar decreased with the column height for all feed sugar contents. The highest effluent ethanol concentration (22.5 g L^?1) and ethanol formation rate were obtained with feed sugar content of 100 g L^?1. Percentage sugar utilization decreased with increasing feed sugar content above 100 g L^?1 yielding lower ethanol contents in the effluent. The highest ethanol yield coefficient (0.52 gE g^?1S) was obtained with a feed sugar content of 50 g L^?1. Biomass concentration also decreased with column height, yielding low ethanol formation in the upper section of the column. CONCLUSION: The packed column bioreactor was found to be effective for ethanol fermentation from CWP solution. The optimum feed sugar content maximizing the effluent ethanol and the specific rate of ethanol formation was found to be 100 g L^?1. High sugar content above 100 g L^?1 resulted in low ethanol productivities due to high maintenance requirements. Copyright © 2008 Society of Chemical Industry     

Statistical optimization of process parameters for the production of vanillic acid by solid‐state fermentation of groundnut shell waste using response surface methodology

BACKGROUND: Vanillic acid is a flavoring agent and also serves as precursor for vanillin production. Culture medium and fermentation condition for the single step production of vanillic acid from Phanerochaete chrysosporium using lignocellulosic waste as a substrate under solid state fermentation (SSF) were optimized using response surface methodology. RESULTS: The process parameters were chosen by borrowing methodology, and L‐asparagine, pH and moisture content of the solid medium during SSF were identified as the most significant variables. The optimum value of selected variable and their mutual interactions were determined by response surface methodology. The result demonstrated that a yield of 73.58 mg vanillic acid g^?1 substate was predicted under optimum conditions (L‐asparagine 5.98 mmol L^?1 (2.37 mg g^?1 groundnut shell), pH of solid medium 4.51 and moisture content 74.83%). The predicted response was experimentally validated and resulted in a maximum vanillic acid yield of 73.69 mg g^?1 after 8 days of SSF. CONCLUSION: The optimization of fermentation variables resulted in a maximum 10‐fold increase in vanillic acid yield compared with that observed under sub‐optimal conditions (from 7.2 mg g^?1 to 73.69 mg g^?1). Copyright © 2011 Society of Chemical Industry     

Detoxification of sugarcane bagasse hemicellulosic hydrolysate with ion‐exchange resins for xylitol production by calcium alginate‐entrapped cells

This study describes the performance of four different resins, in sequence, to detoxify sugarcane bagasse hemicellulosic hydrolysate and to improve xylitol production by calcium alginate‐entrapped Candida guilliermondii FTI20037 cells under conditions of low oxygen concentration. The treatment resulted in a removal of 82.1% furfural, 66.5% hydroxymethylfurfural, 61.9% phenolic compounds derived from lignin degradation, 100% chromium, 46.1% zinc, 28.5% iron, 14.7% sodium and 3.5% nickel. On the other hand, the removal of acetic acid was not significant. A xylitol yield factor (Y_P/S) of 0.62 g g^?1 and a volumetric productivity (Q_p) of 0.24 g dm^?3 h^?1 were attained in the fermentation process for xylitol production from detoxified hydrolysate. Copyright © 2004 Society of Chemical Industry     

Sorption capacity of a new generation granular activated carbon—Bio‐Sep® bead—and its use in a suspended growth bioreactor

BACKGROUND: A new generation granular activated carbon—Bio‐Sep® beads—consist of 25% polymer (Nomex) and 75% powdered activated carbon. The porous structure and high surface area of these beads make them suitable for sorbent in adsorption columns, and for immobilization media in bioreactors. The aim of this study was to study the sorption characteristics of Bio‐Sep® beads for methyl t‐butyl ether (MTBE) and t‐butyl alcohol (TBA), and to demonstrate the advantage of their usage in a suspended growth bioreactor. RESULTS: The maximum uptake capacity of Bio‐Sep® beads for MTBE and TBA, in the studied concentration range (10–100 mg L^?1), was observed to be 9.73 and 6.23 mg g^?1, respectively. A 52 h desorption experiment resulted in 13.6–42.2% MTBE and 33–53% TBA desorption corresponding to the initial solid phase concentrations of 1.68–9.73 mg g^?1 and 1.41–6.23 mg g^?1, respectively. The sorption of TBA on the Bio‐Sep® beads was significantly hindered by the presence of MTBE. The addition of 10 g Bio‐Sep® beads (dry weight) in a suspended growth bioreactor was able to eliminate the inhibitory effect of 150 mg L^?1 MTBE. CONCLUSIONS: At an equilibrium aqueous phase concentration (C_e) of 1 mg L^?1, the solid phase concentration (q_e) on Bio‐Sep® beads were observed as 1.44 and 0.47 mg g^?1 for MTBE and TBA, respectively. The results obtained in this study indicate that Bio‐Sep® beads have reasonable sorption and desorption characteristics, which can be successfully exploited for the removal/degradation of toxic organic pollutants in high rate bioreactors. Copyright © 2007 Society of Chemical Industry     

Improvement of heat removal in solid‐state fermentation tray bioreactors by forced air convection

BACKGROUND: Heat removal is one of the major constraints in large‐scale solid‐state fermentation (SSF) processes. The effect of internal air circulation by forced convection on heat and water transfer has not been studied in SSF tray bioreactors. Formulation of a mathematical model for SSF requires a good estimation of the mass and heat transfer coefficients. RESULTS: A stainless steel tray bioreactor (80.6 L capacity) was used. Aspergillus niger C28B25 was cultivated under SSF conditions on an inert support. Temperature, moisture content, biomass and substrate concentrations were measured. Water and energy integral balances were used to estimate the heat and mass transfer coefficients involved in the process. The Reynolds number (N_Re) in the headspace of the tray bioreactor ranged from 2.5 to 2839, which increased the global heat transfer coefficient from 4.2 to 6.9 (W m^?2 K^?1) and the mass transfer coefficient from 1.0 to 2.1 (g m^?2 s^?1). Mathematical model predictions of the temperature and moisture content of the fermentation bed showed a high goodness‐of‐fit with the experimental results. CONCLUSIONS: This is the first report describing the effect of N_Re of air in the headspace of a SSF tray bioreactor on the heat and mass transfer coefficients and temperature regulation in SSF. Copyright © 2011 Society of Chemical Industry     

Ni(ll) biosorption by Rhizopus arrhizus Env 3: the study of important parameters in biomass biosorption

BACKGROUND: The removal of toxic metals from wastewaters by biosorption, based on the metal‐binding capacities of various biological materials, has attracted much interest. However, the success of this approach depends on economic feasibility, which can be obtained by optimisation of the environmental conditions. In this study, Ni(II) biosorption experiments were carried out using a preformed biomass of Rhizopus arrhizus. A pure culture of previously isolated R. arrhizus Env 3 was used for maximum biosorption of nickel metal from nickel‐electroplating industrial effluent. RESULTS: Various environmental factors such as nickel concentration, pH, temperature, mycelial pellet weight, pretreatment of fungal biomass, dead and living fungal biomass and time course of biosorption by R. arrhizus Env 3 were optimised for maximum removal of nickel from the effluent. The maximum nickel removal rate of 618.5 mg g^?1 was observed with living biomass at pH 8, temperature 35 °C, nickel concentration 500 mg L^?1, pellet size 3 g wet weight and shaker velocity 150 rpm. Maximum nickel biosorption was obtained after 72 h. CONCLUSION: Statistical analysis of different factors such as temperature, pH, mycelial pellet size, concentration of nickel in effluent and residual nickel level showed that all these factors had significant effects on the biosorption of nickel metal by R. arrhizus Env 3 from nickel‐electroplating industrial effluent. Copyright © 2008 Society of Chemical Industry     

Improved mass transfer and biodegradation rates of naphthalene particles using a novel bead mill bioreactor

One of the main challenges in the treatment of polycyclic aromatic hydrocarbons (PAHs) in controlled bioreactors is the hydrophobicity and low solubility of these compounds in the aqueous phase, resulting in appreciable mass transfer limitations within the bioreactor. To address this challenge, we have developed a modified roller bioreactor (Bead Mill Bioreactor) in which inert particles are used to improve mass transfer from the solid phase to the aqueous phase. Experimental results with naphthalene as a model PAH and Pseudomonas putida as a candidate bacterium indicate that both the mass transfer rate from the solid phase to liquid phase and the biodegradation rate in the Bead Mill Bioreactor (BMB) were significantly higher than those in a conventional roller bioreactor (20‐fold and 5.5‐fold, respectively). The enhancement of mass transfer was dependent on the type, size and volumetric loading of the inert particles, as well as concentration of particulate naphthalene. The highest mass transfer coefficient (k_La = 2.1 h^?1) was achieved with 3 mm glass beads at a volumetric loading of 50% (particle volume/working volume) with 10 000 mg dm^?3 particulate naphthalene. The maximum biodegradation rate of naphthalene attained in the bead mill bioreactor (59.2 mg dm^?3 h^?1 based on the working volume and 118.4 mg dm^?3 h^?1 based on the liquid volume) surpasses most other rates published in the literature and is equivalent to values reported for more complex bioreaction systems. The bead mill bioreactor developed in the present work not only enjoys a simple design but shows excellent performance for treatment of PAHs suspended in an aqueous phase. This fundamental information will be of significant value for future studies involving soil‐bound PAHs. Copyright © 2005 Society of Chemical Industry     

Xylitol production by Candida tropicalis from corn cob hemicellulose hydrolysate in a two‐stage fed‐batch fermentation process

BACKGROUND: Xylitol, a sugar alcohol widely used in food and pharmaceutical industries, can be produced through biological reduction of xylose present in hemicellulose hydrolysates by Candida tropicalis. However, the aeration rate and by‐products originating from hemicellulose hydrolysis strongly inhibit the production of xylitol in a fermentation process. A two‐stage fed‐batch fermentation system was developed to reduce these inhibitory effects and to improve xylitol production from corn cob hemicellulose hydrolysates by C. tropicalis. RESULTS: Results of batch fermentations indicated that high xylitol production could be obtained from C. tropicalis at an initial xylose concentration of 80 g L^?1 in corn cob hydrolysate medium at an aeration rate of 0.4 vvm at the micro‐aeration stage. In the two‐stage fed‐batch fermentation process, 96.5 g L^?1 xylitol was obtained after 120 h, giving a yield of 0.83 g g^?1 and a productivity of 1.01 g L^?1 h^?1, which were 12.16% and 65.57% higher than those in a batch fermentation. CONCLUSION: High xylitol production can be achieved in a two‐stage fed‐batch fermentation process, in which the negative effects of aeration rate and inhibitory compounds on xylitol formation can be considerably reduced. Copyright © 2011 Society of Chemical Industry     

Fermentative production of fumaric acid from Eucalyptus globulus wood hydrolyzates

Fumaric acid (FA) was produced from Eucalyptus globulus wood by successive steps of hydrothermal processing (to solubilize hemicelluloses and to increase the susceptibility of solids to enzymatic hydrolysis), enzymatic hydrolysis and fermentation with Rhizopus arrhizus DSM 5772. For comparative purposes, additional fermentations were carried out using synthetic media. Single stage fermentation of synthetic media led to a medium containing 11.8 g FA L^?1 (Y_P/S = 0.60 g g^?1). Operating in fed batch mode, the fourth stage increased the FA concentration from 19.7 up to 43.6 g L^?1 (Y_P/S = 0.71 g g^?1). Hydrolyzate fermentation in a single stage resulted in lower fumaric acid concentration (9.65 g L^?1) and yield (0.35 g g^?1). Additional fermentations were carried out in media made with hydrolyzates subjected to membrane processing, adsorption or ion exchange. The highest yield (Y_P/S = 0.44 g g^?1) was reached in media made up of ion‐exchange treated hydrolyzates and a commercial glucose solution in proportion 85/15 w/w. Copyright © 2011 Society of Chemical Industry     

Inoculum strategies for Penicillium simplicissimum lipase production by solid‐state fermentation using a residue from the babassu oil industry

Two alternative inoculation strategies for lipase production by the fungus Penicillium simplicissimum were tested in solid‐state fermentation using a residue from the babassu oil industry (babassu cake). Conventional spore inoculation was compared with fungal pellets grown in liquid medium and with inocula consisting of fermented cake. Fungal pellets delayed lipase production whereas fermented cake accelerated enzyme synthesis, yielding a productivity of 0.45 U g^?1 h^?1, which is equivalent to the highest values obtained with conventional inocula. Therefore, a 2² factorial design was used to determine the best conditions for lipase production with fermented cake as inoculum strategy, varying the inoculum propagation time and inoculum concentration. Lipase activity and productivity reached 30 U g^?1 and 0.63 U g^?1 h^?1, respectively, with 10% inoculum and 36 h. Thus, fermented cake inocula increased production 1.5‐fold with 10 times fewer spores than in conventional inoculation, indicating that fermented solids are an interesting alternative for inoculum development in solid‐state fermentation, mainly for large‐scale processes. Copyright © 2007 Society of Chemical Industry     

Time‐dependent dosing of Fe2+ for improved lipopeptide production by marine Bacillus megaterium

BACKGROUND: Lipopeptide production is strongly influenced by trace metals. The availability of free Fe²⁺ in the media throughout the process of fermentation was found to be very critical. Since free Fe²⁺ was reported to be sequestered by the lipopeptide as it was produced, intermittent feeding of Fe²⁺ was strategized and optimized for enhanced lipopeptide production by marine Bacillus megaterium in glucose mineral salts medium (GMSM). RESULTS: Studies with the single‐dose Fe²⁺ (0.48 mmol L^?1) supplementation after 8 h of fermentation resulted in lipopeptide concentration of 3.3 ± 0.1 g L^?1. Lipopeptide production was further enhanced to 4.2 ± 0.15 g L^?1 by adopting a multi‐dose Fe²⁺ feeding strategy. The maximum product yield (Y_P/S) of 0.24 ± 0.02 g g^?1 with specific product formation rate (q_p) of 0.124 ± 0.01 g g^?1 h^?1 was achieved when 0.48 mmol L^?1 Fe²⁺ was fed intermittently at different times as per the designed strategy. CONCLUSION: Lipopeptide concentration was improved 4.7‐fold by single‐dosing and 5.8‐fold by multiple dosing of Fe²⁺, when compared with GMSM without Fe²⁺ supplementation. Copyright © 2012 Society of Chemical Industry     

Optimization of pullulan production using Ca‐alginate‐immobilized Aureobasidium pullulans by response surface methodology

BACKGROUND: The production of pullulan from synthetic medium by Aureobasidium pullulans P56 immobilized in Ca‐alginate beads was investigated using batch and repeated batch fermentation systems. RESULTS: The highest pullulan concentration (19.52 ± 0.37 g dm^?3) was obtained with 2.0‐2.4 mm beads prepared from 2% sodium alginate solution. Pullulan production was mainly accomplished by immobilized fungal cells since leaked cells in the fermentation medium comprised 17.4% of the total fungal population at the end of fermentation. The pullulan proportion was 84.5% of the total polysaccharide in the fermentation medium. Response surface methodology was used to investigate the effects of three fermentation parameters (initial pH, agitation speed and incubation time) on the concentration of pullulan. Results of the statistical analysis showed that the fit of the model was good in all cases. The maximum pullulan concentration of 21.07 ± 0.48 g dm^?3 was obtained at the optimum concentrations of process variables (pH 7.31, agitation speed 191.5 rpm, incubation time 101.2 h). The gel beads produced pullulan under the optimized conditions for six consecutive batch fermentations without marked activity loss and deformation. CONCLUSION: The results of this study suggest that the immobilization of A. pullulans cells in Ca‐alginate gel beads is suitable for batch and repeated batch production of pullulan. Copyright © 2007 Society of Chemical Industry