Optimization of chitosanase production from Bacillus sp. RKY3 using statistical experimental designs

BACKGROUND: The culture medium and fermentation conditions for the production of constitutive chitosanase from a newly isolated Bacillus sp. RKY3 were optimized statistically. RESULTS: The variables significantly influencing both chitosanase production and cell growth were screened through the Plackett–Burman design, by which maltose, beef extract, MgSO₄, and incubation time were identified as the most significant variables. The optimum values of the selected variables and their mutual interactions were determined through the steepest ascent method and Box–Behnken experimental design. The results demonstrated that 62.30 U mL^?1 chitosanase activity was predicted with optimum conditions of maltose (30.18 g L^?1), beef extract (15.25 g L^?1), MgSO₄ (0.26 g L^?1), and incubation time (50.02 h). The predicted response was verified by the validation experiments, and the optimum conditions resulted in a maximum chitosanase activity of 63.53 ± 1.22 U mL^?1. CONCLUSION: The optimization of fermentation variables resulted in an approximately 11.3‐fold increase in chitosanase activity compared with that observed under unoptimized conditions (from 5.63 U mL^?1 to 63.53 U mL^?1). Copyright © 2009 Society of Chemical Industry     

Production of organic solvent tolerant lipase by Staphylococcus caseolyticus EX17 using raw glycerol as substrate

BACKGROUND: In this work we used Plackett–Burman statistical design and central composite design in order to optimize culture conditions for lipase production by Staphylococcus caseolyticus strain EX17 growing on raw glycerol, which was obtained as a by‐product of the enzymatic synthesis of biodiesel. The stability of lipase was verified over several organic solvents, such as methanol, ethanol and n‐hexane. RESULTS: Optimal culture conditions for lipase production were found to be 36 °C, initial pH 8.12, glycerol 30 g L^?1, olive oil 3.0 g L^?1, and soybean oil 2.5 g L^?1, with 145.8 U L^?1 of enzyme activity. When commercial glycerol was substituted by the raw glycerol from biodiesel synthesis, lipolytic activity was 127.3 U L^?1. Experimental validation of enzyme production matched values predicted by the mathematical model, which was 138.3 U L^?1. Stability tests showed that lipase from S. caseolyticus EX17 was stable in methanol, ethanol, and n‐hexane. CONCLUSIONS: Results obtained in this work suggest that raw glycerol can be used for lipase production by S. caseolyticus EX17 and that this enzyme has a potential application in the synthesis of biodiesel. Copyright © 2008 Society of Chemical Industry     

Optimization of alkaline protease production from Shewanella oneidensis MR‐1 by response surface methodology

BACKGROUND: The proteases are among the most important groups of enzymes. Therefore, it is important to produce inexpensive and optimized media for large‐scale commercial production. In the present work, three different Shewanella species were screened on skim milk agar medium for their ability to produce alkaline protease. The effects of different culture conditions were optimized for alkaline protease production by S. oneidensis MR‐1 using a Box–Behnken design combined with response surface methodology (RSM). RESULTS: Highest yield (112.90 U mL^?1) of protease production was obtained at pH 9.0, a temperature of 30 °C, glucose (12.5 g L^?1), tryptone (12.5 g L^?1) and an incubation period of 36 h. A second‐order polynomial regression model was used for analysis of the experiment. The experimental values were in good agreement with predicted values, with correlation coefficient 0.9996. CONCLUSION: Carbon and nitrogen, pH, temperature and incubation period were chosen as the main factors to be used in an experimental design for optimization to produce low‐cost enzymes, potentially for use on an industrial scale. A 60% increase in enzyme activity was achieved in the optimized medium compared with the original medium. Copyright © 2008 Society of Chemical Industry     

Conversion efficiency of glucose/xylose mixtures for ethanol production using Saccharomyces cerevisiae ITV01 and Pichia stipitis NRRL Y‐7124

BACKGROUND: Efficient conversion of glucose/xylose mixtures from lignocellulose is necessary for commercially viable ethanol production. Oxygen and carbon sources are of paramount importance for ethanol yield. The aim of this work was to evaluate different glucose/xylose mixtures for ethanol production using S. cerevisiae ITV‐01 (wild type yeast) and P. stipitis NRRL Y‐7124 and the effect of supplying oxygen in separate and co‐culture processes. RESULTS: The complete conversion of a glucose/xylose mixture (75/30 g L^?1) was obtained using P. stipitis NRRL Y‐7124 under aerobic conditions (0.6 vvm), the highest yield production being Y_p/s = 0.46 g g^?1, volumetric ethanol productivity Q_pmax = 0.24 g L^?1 h^?1 and maximum ethanol concentration P_max = 34.5 g L^?1. In the co‐culture process and under aerobic conditions, incomplete conversion of glucose/xylose mixture was observed (20.4% residual xylose), with a maximum ethanol production of 30.3 g L^?1, ethanol yield of 0.4 g g^?1 and Q_pmax = 1.26 g L^?1 h^?1. CONCLUSIONS: The oxygen present in the glucose/xylose mixture promotes complete sugar consumption by P. stipitis NRRL Y‐7124 resulting in ethanol production. However, in co‐culture with S. cerevisiae ITV‐01 under aerobic conditions, incomplete fermentation occurs that could be caused by oxygen limitation and ethanol inhibition by P. stipitis NRRL Y‐7124; nevertheless the volumetric ethanol productivity increases fivefold compared with separate culture. Copyright © 2011 Society of Chemical Industry     

Kinetic study on ethanol production using Saccharomyces cerevisiae ITV‐01 yeast isolated from sugar cane molasses

BACKGROUND: Bio‐ethanol production from renewable sources, such as sugar cane, makes it a biofuel that is both renewable and environmentally friendly. One of the strategies to reduce production costs and to make ethanol fuel economically competitive with fossil fuels could be the use of wild yeast with osmotolerance, ethanol resistance and low nutritional requirements. The aim of this work was to investigate the kinetics of ethanol fermentation using Saccharomyces cerevisiae ITV‐01 yeast strain in a batch system at different glucose and ethanol concentrations, pH values and temperature in order to determine the optimum fermentation conditions. RESULTS: This strain showed osmotolerance (its specific growth rate (µ_max) remained unchanged at glucose concentrations between 100 and 200 g L^?1) as well as ethanol resistance (it was able to grow at 10% v/v ethanol). Activation energy (Ea) and Q₁₀ values calculated at temperatures between 27 and 39 °C, pH 3.5, was 15.6 kcal mol^?1 (with a pre‐exponential factor of 3.8 × 10¹² h^?1 (R² = 0.94)) and 3.93 respectively, indicating that this system is biologically limited. CONCLUSIONS: The optimal conditions for ethanol production were pH 3.5, 30 °C and initial glucose concentration 150 g L^?1. In this case, a maximum ethanol concentration of 58.4 g L^?1, ethanol productivity of 1.8 g L^?1 h^?1 and ethanol yield of 0.41 g g^?1 were obtained. Copyright © 2010 Society of Chemical Industry     

Sequential optimization of culture medium composition for extracellular lipase production by Bacillus sphaericus using statistical methods

A sequential optimization strategy with the aid of statistical design of experiments was used to enhance the lipase (triacylglycerol acylhydrolases, EC 3.1.1.3) production by Bacillus sphaericus in submerged cultivation. A Plackett–Burman experimental design was used to evaluate the twelve medium components. Various vegetable oil inducers were tested for lipase production in the second step and the third step was to identify the optimal values of the significant medium components with sesame oil as the inducer using response surface methodology. A predictive model of the combined effects of the independent variables using response surface methodology and an artificial neural network was proposed. Unstructured kinetic models, a logistic model and a Luedeking–Piret model, were used to describe the cell mass and lipase production respectively. The significant variables affecting lipase production were found to be glucose, olive oil, peptone, NaCl and MnSO₄.H₂O. Sesame oil was found to be the best inducer for lipase production by Bacillus sphaericus. The maximum lipase activity of 4.45 U mL^?1, which was 1.5 times the maximum activity obtained in the Plackett–Burman experimental trials, was obtained at the optimum combination of medium constituents containing 12.695 g L^?1 glucose, 13.161 mL L^?1 sesame oil, 9.947 g L^?1 peptone, 3.25 g L^?1 NaCl, 0.5917 g L^?1 MnSO₄.H₂O and other insignificant components at the fixed level. The statistical design of experiments offers an efficient methodology to identify the significant variables and to optimize the factors with a minimum number of experiments for lipase production by Bacillus sphaericus. Copyright © 2007 Society of Chemical Industry     

Potential fuel oils from the microalga Choricystis minor

BACKGROUND: Continuous culture of the freshwater microalga Choricystis minor was investigated for possible use in producing lipid feedstock for making biofuels. The effects of temperature (10–30 °C) and dilution rate (0.005–0.017 h^?1) on lipid productivity in a nutrient sufficient medium in a 4 L stirred tank bioreactor under continuous illumination at an incident irradiance level of 550 µE · m^?2s^?1 and a controlled pH of 6 under carbon dioxide supplemented conditions are reported. RESULTS: The maximum lipid productivity was 82 mg L^?1 d^?1 at 25 °C and a dilution rate of 0.014 h^?1. Lipid contents of the biomass were 21.3 ± 1.7 g per 100 g of dry biomass, irrespective of the culture temperature and dilution rate. After the biomass had been grown in nutrient sufficient conditions in continuous culture, it was recovered and subjected to various postharvest treatments. With the best postharvest treatment, the neutral lipid contents of the algal biomass were raised ～6‐fold relative to untreated biomass. CONCLUSION: At 82 mg L^?1 d^?1, or 21 000 L ha^?1 year^?1, the lipid productivity of C. minor was nearly four times the lipid productivity of oil palm, a highly productive crop. Therefore, C. minor is potentially a good source of renewable lipid feedstock for biofuels. Copyright © 2009 Society of Chemical Industry     

Modulated gluconic acid production from immobilized cells of Aspergillus niger ORS‐4.410 utilizing grape must

BACKGROUND: Gluconic acid (GA) production by immobilized cells of mutant Aspergillus niger ORS‐4.410 on polyurethane sponge (PUS) and calcium‐alginate (Ca‐alginate) was evaluated in repeated batches of solid state surface fermentation (SSF) and submerged fermentation (SmF) conditions, respectively, utilizing rectified grape must as carbon source. RESULTS: The passive immobilization of cells in fermentation medium solid support of having 0.4 cm³ cube size, 4% spore suspension, 0.6 g inoculum of PUS immobilized cells at 32 °C and 2.0 L min^?1 resulted in the maximum GA production (88.16 g L^?1) with a 92.8% yield, while the Ca‐alginate matrix with a 0.5 cm diameter bead size, 2–3% spore suspension, 15 g inoculum at 34 °C and 150 rpm agitation speed revealed 67.19 g L^?1 GA with a 85.2% yield. Repeated use of PUS showed higher levels of GA (110.94 g L^?1) in the third–fourth fermentation cycles with 95–98% yield and 22.50 g L^?1 d^?1 productivity under SSF that was 2.5‐fold higher than the productivity obtained from a typical fermentation cycle, and 54% greater than the productivity obtained with repetitive use of Ca‐alginate immobilized cells of A. niger under SmF. CONCLUSION: Using immobilized cells of A. niger in PUS, the rectified form of grape must can be utilized for GA production as an alternative source of carbohydrate by replacing the conventional fermentation conditions. Copyright © 2008 Society of Chemical Industry     

Effect of acetic acid on lipid accumulation by glucose‐fed activated sludge cultures

BACKGROUND: The effect of acetic acid, a lignocellulose hydrolysis by‐product, on lipid accumulation by activated sludge cultures grown on glucose was investigated. This was done to assess the possible application of lignocellulose as low‐cost and renewable fermentation substrates for biofuel feedstock production. RESULTS: Biomass yield was reduced by around 54% at a 2 g L^?1 acetic acid dosage but was increased by around 18% at 10 g L^?1 acetic acid dosage relative to the control run. The final gravimetric lipid contents at 2 and 10 g L^?1 acetic acid levels were 12.5 ± 0.7% and 8.8 ± 3.2% w/w, respectively, which were lower than the control (17.8 ± 2.8% w/w). However, biodiesel yields from activated sludge grown with acetic acid (5.6 ± 0.6% w/w for 2 g L^?1 acetic acid and 4.2 ± 3.0% w/w for 10 g L^?1 acetic acid) were higher than in raw activated sludge (1–2% w/w). The fatty acid profiles of the accumulated lipids were similar with conventional plant oil biodiesel feedstocks. CONCLUSIONS: Acetic acid enhanced biomass production by activated sludge at high levels but reduced lipid production. Further studies are needed to enhance acetic acid utilization by activated sludge microorganisms for lipid biosynthesis. Copyright © 2011 Society of Chemical Industry     

Strain isolation and optimization of process parameters for bioconversion of glycerol to lactic acid

BACKGROUND: The crude glycerol from biodiesel production represents an abundant and inexpensive source which can be used as raw material for lactic acid production. The first aim of this investigation was to select a strain suitable for producing lactic acid from glycerol with a high concentration and productivity. The second aim was to obtain the optimum fermentation conditions, as a basis for large‐scale lactate production in the future. RESULTS: Eight bacterial strains, which could aerobically convert glycerol to lactic acid, were screened from soil samples. One of the strains, AC‐521, which synthesized lactic acid with a higher concentration, was identified based on its 16S rDNA sequences and physiological characteristics. These results indicated that this strain was a member of Escherichia coli. The optimal fermentation conditions for Escherichia coli AC‐521 were 42 °C, pH 6.5, 0.85 min^?1 (K_La). CONCLUSION: Escherichia coli AC‐521 suitable for producing lactic acid from glycerol with high concentration and productivity was identified. After 88 h of fed‐batch fermentation, both the lactic acid concentration and glycerol consumption reached maximum, giving 85.8 g L^?1 of lactic acid with a productivity of 0.97 g L^?1 h^?1 and a yield of 0.9 mol mol^?1 glycerol. Copyright © 2009 Society of Chemical Industry     

Characterization of a novel glutamate decarboxylase from Streptococcus salivarius ssp. thermophilus Y2

BACKGROUND: γ‐Aminobutyric acid with several well‐known physiological functions is biosynthesized via the irreversible α‐decarboxylation of L ‐glutamate catalysed by glutamate decarboxylase (GAD). Although Streptococcus salivarius ssp. thermophilus has been widely applied to the dairy, the characterization of its GAD has not been reported. In this paper, the purification and the characterization of S. salivarius ssp. thermophilus GAD were investigated. RESULTS: GAD was purified 22‐fold from crude protein extracts with a yield of 7.8% in five steps. The final preparation gave a single band on SDS‐PAGE. The molecular weight of GAD determined by SDS‐PAGE and gel filtration was 46.9 kDa and 103.6 kDa, respectively, indicating that the enzyme exists as a dimmer of homological subunits. The optimum temperature and pH of GAD was 55 °C and pH 4.0, respectively. The enzyme reacted only with L ‐glutamate among 19 α‐amino acids with apparent K_m at 2.3 mmol L^?1 and did not react with D ‐glutamic acid. Activity of the enzyme could significantly be activated by 5 mmol L^?1 of BaCl₂ and inhibited by FeSO₄, ZnSO₄, CuSO₄, MnSO₄, Na₂SO₄, AgNO₃, CoCl₂, LiCl and KCl, respectively. The N‐terminal amino acid sequence of GAD was NH₂‐MNEKLFREI. CONCLUSION: Both the characterization and the deduced amino sequence (ABI31651) showed the purified enzyme was a novel GAD. Copyright © 2008 Society of Chemical Industry     

Effective biosynthesis of poly(3‐hydoxy‐ butyrate‐co‐4‐hydroxybutyrate) with high 4‐hydroxybutyrate fractions by Wautersia eutropha in the presence of α‐amino acids

BACKGROUND: Biopolymers produced by microbes are in demand as their biodegradable and biocompatible properties make them suitable for disposable products and for potential use as biomaterials for medical applications. The effective microbial production of copolyesters of 3‐hydroxybutyrate (3HB) and 4‐hydroxybutyrate(4HB) with high molar fractions of 4HB unit by a wild‐type Wautersia eutropha H16 was investigated in culture media containing 4‐hydroxybutyric acid (4HBA) and different carbon substrates in the presence of various α‐amino acids. RESULTS: The addition of carbon sources such as glucose, fructose and acetic acid to the culture medium containing 4HBA in the presence of α‐amino acids resulted in the production of random poly(3HB‐co‐4HB) with compositions of up to 77 mol% 4HB unit, but the yields of copolyesters with 60–77 mol% 4HB units were less than 15 wt% of dried cell weights. In contrast, when carbon sources such as propionic acid and butyric acid were used as the co‐substrates of 4HBA in the presence of α‐amino acids, poly(3HB‐co‐4HB) copolyesters with compositions of 72–86 mol% 4HB were produced at maximally 47.2 wt% of dried cell weight (11.3 g L^?1) and the molar conversion yield of 4HBA to 4HB fraction in copolyesters was as high as 31.4 mol%. Further, poly(3HB‐co‐4HB) copolyesters with compositions of 93–96 mol% 4HB were isolated at up to 35.2 wt% of dried cell weights by fractionation of the above copolymers with chloroform/n‐hexane. CONCLUSION: The productivity of copolyesters with over 80 mol% 4HB fractions was as high as 0.146 g L^?1 h^?1 (3.51 g L^?1 for 24 h) by flask batch cultivation. Copyright © 2007 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     

Efficient acetoin production by optimization of medium components and oxygen supply control using a newly isolated Paenibacillus polymyxa CS107

BACKGROUND: Acetoin is a natural flavor commonly used in wine, buffer, honey, garnet berry and strawberry as a food additive. It also has been widely applied in cosmetics, pharmacy and chemical synthesis. Culture medium optimization and process control were carried out for efficient production of acetoin by a newly isoliated P. polymyxa CS107. RESULTS: An acetoin high producing strain, designated as CS107, was newly isolated and identified as P. polymyxa based on its physiological and biochemical characteristics as well as the 16S rDNA sequence. The medium composition was optimized in shake flask fermentations by a sequential statistical experimental design. Under the optimized conditions, acetoin concentration of 30.98 g L^?1 was achieved with 71.83% of theoretical glucose conversion efficiency. Fed‐batch fermentation based on a suitable agitation speed was carried out in a 5 L jar, the maximum acetoin concentration of 55.3 g L^?1 was obtained with the productivity of 1.32 g L^?1 h^?1 and the yield of 75.62%. CONCLUSION: A new strain for efficient production of acetoin, designated as P. polymyxa CS107, was obtained. The optimization of fermentation variables and fed‐batch culture resulted in a maximum acetoin concentration of 55.3 g L^?1 in 5 L jar. Copyright © 2012 Society of Chemical Industry     

Biomonitoring of biosurfactant production by green fluorescent protein‐marked Bacillus subtilis W1012

BACKGROUND: Biosurfactant production was investigated using two strains of Bacillus subtilis, one being a reference strain (B. subtilis 1012) and the other a recombinant of this (B. subtilis W1012) made able to produce the green fluorescent protein (GFP). RESULTS: Batch cultivations carried out at different initial levels of glucose (G₀) in the presence of 10 g L^?1 casein demonstrated that the reference strain was able to release higher levels of biosurfactants in the medium at 5.0≤G₀≤10 g L^?1 (B_max = 104–110 mg L^?1). The recombinant strain exhibited slightly lower levels of biosurfactants (B_max = 90–104 mg L^?1) but only at higher glucose concentrations (G₀ ≥ 20 g L^?1). Under these nutritional conditions, the fluorescence intensity linked to the production of GFP was shown to be associated with the cell concentration even after achievement of the stationary phase. CONCLUSION: The ability of the genetically‐modified strain to simultaneously overproduce biosurfactant and GFP even at low biomass concentration makes it an interesting candidate for use as a biological indicator to monitor indirectly the biosurfactant production in bioremediation treatments. Copyright © 2008 Society of Chemical Industry     

Batch cooling crystallization of xylitol produced by biotechnological route

BACKGROUND: This work deals with the xylitol production by biotechnological routes emphasizing the purification process using crystallization. RESULTS: Xylitol volumetric productivity of 0.665 g L^?1 h^?1 and yield of 0.7024 g g^?1 were obtained after 92 h fermentation. The fermented broth (61.3 g L^?1 xylitol) was centrifuged, treated and concentrated obtain a syrup (745.3 g L^?1 xylitol) which was crystallized twice, xylitol crystals with 98.5–99.2% purity being obtained. CONCLUSION: The hypothetical distribution obtained permits the determination of modeling parameters, which make possible the estimation of crystal dominant size from different initial experimental conditions. Copyright © 2008 Society of Chemical Industry     

Production of fructooligosaccharides by β‐fructofuranosidase from Aspergillus sp 27H

β‐fructofuranosidase (EC 3.2.1.26) from Aspergillus sp 27H isolated from soil was investigated for production of fructooligosaccharides (FOS) using whole cells. It possesses hydrolytic and transfructosylating activities that can be altered by modifying the reaction conditions. The optimal conditions for the transfructosylating activity occur in the pH range 5.5–6.0 and at 60 °C, while hydrolytic activity was highest at pH 4.0 and 55 °C. At low sucrose concentration (10 g dm^?3) there was rapid conversion of sucrose to glucose and fructose and very low concentrations of FOS were obtained. However, at sucrose concentrations higher than 216 g dm^?3 the concentrations of hydrolysis products were reduced. Under the following conditions: pH 5.5, temperature 40 °C, sucrose concentration 615 g dm^?3 and enzyme concentration 20β‐fructofuranosidase units g^?1 of sucrose, the FOS concentration reached a maximum value of 376 g dm^?3 (234 g dm^?3 1‐kestose and 142 g dm^?3 nystose) and the proportion of FOS in the solids in the reaction mixture was 600–620 g kg^?1 at 6 h. These results suggest that β‐fructofuranosidase from Aspergillus sp 27H could be an appropriate enzyme for the commercial production of FOS. Copyright © 2004 Society of Chemical Industry     

Continuous lipase‐catalyzed synthesis of hexyl laurate in a packed‐bed reactor: optimization of the reaction conditions in a solvent‐free system

BACKGROUND: Hexyl laurate has been applied widely in cosmetic industries and is synthesized by chemical methods with problems of cost, environmental pollution, and by‐products. In this study, Lipozyme^® IM77 (from Rhizomucor miehei) was used to catalyze the direct‐esterification of hexanol and lauric acid in a solvent‐free system by utilizing a continuous packed‐bed reactor, wherein the aforementioned difficulties could be overcome. Response surface methodology (RSM) and three‐level‐three‐factor Box‐Behnken design were employed to evaluate the effects of synthesis parameters, such as reaction temperature (45–65 °C), mixture flow rate (0.25–0.75 mL min^?1) and concentration of lauric acid (100–300 mmol L^?1) on the production rate (µmol min^?1) of hexyl laurate by direct esterification. RESULTS: The production rate was affected significantly by the mixture flow rate and lauric acid concentration. On the basis of ridge‐max analysis, the optimum synthesis conditions for hexyl laurate were as follows: 81.58 ± 1.76 µmol min^?1 at 55 °C, 0.5 mL min^?1 flow rate and 0.3 mol L^?1 lauric acid. CONCLUSION: The lipase‐catalyzed synthesis of hexyl laurate by Lipozyme^® IM‐77 in a continuous packed‐bed bioreactor and solvent‐free system was successfully developed; optimization of the reaction parameters was obtained by Box–Behnken design and RSM. Copyright © 2008 Society of Chemical Industry     

Continuous production of poly‐β‐hydroxybutyrate by high‐cell‐density cultivation of Wautersia eutropha

BACKGROUND: Poly‐β‐hydroxybutyrate (PHB) accumulation is triggered by limitation of a nutrient other than carbon. The production cost of PHB is very high. In order to reduce this cost, continuous cultivation for the accumulation of PHB was investigated. The culture was first allowed to grow under fed‐batch conditions to yield a significant increase in biomass and PHB accumulation. Thereafter this high‐cell‐density biomass containing PHB was allowed to grow and maintained under conditions of continuous cultivation so that the overall process could be simplified and economised. RESULTS: For continuous cultivation a medium containing 90 g L^?1 fructose and 2.5 g L^?1 nitrogen (as urea) was fed continuously at a dilution rate of 0.1 h^?1. A steady state biomass of 27.7 g L^?1 with a PHB concentration of 5.5 g L^?1 was established in the bioreactor. This resulted in a continuous PHB productivity of 0.55 g L^?1 h^?1. CONCLUSION: The experiments have resulted in the development of a novel production technology involving the integration of batch, fed‐batch and continuous processes. At the same time the production of PHB under continuous cultivation increases the overall industrial importance of the system. Copyright © 2008 Society of Chemical Industry     

The use of olive mill wastewater by wild type Yarrowia lipolytica strains: medium supplementation and surfactant presence effect

BACKGROUND: The aim of this work was to study the ability of two different wild type strains of the yeast Yarrowia lipolytica to grow on olive mill wastewater (OMW) and their potential to produce high‐value products such as lipases. Factors that affect cellular growth and OMW degradation were studied, such as nitrogen supplementation, cells concentration and surfactant addition. RESULTS: Both strains, W29 and IMUFRJ 50682, were able to grow in OMW with 19 g L^?1 of COD and approximately 800 mg L^?1 of total phenols concentration. The strain W29 presented the highest potential for extracellular lipase production in OMW medium. Lipase productivity was improved by the medium supplementation with ammonium sulphate up to 6 g L^?1, leading to 80% of COD degradation and 70% of total phenols reduction. The surfactant Tween 80 enhanced cell growth and COD degradation, but had a negative effect on lipase activity. CONCLUSIONS Y. lipolytica has a great potential for OMW valorisation by its use as culture medium for biomass and enzymes production. The operating conditions that favoured lipase production differ from the conditions that improve COD reduction. Copyright © 2008 Society of Chemical Industry