Production of L‐lactic acid with very high gravity distillery wastewater from ethanol fermentation by a newly isolated Enterococcus hawaiiensis

BACKGROUND: A great amount of wastewater with high contents of chemical oxygen demand (COD) are produced by ethanol production. It would be useful to utilize distillery wastewater to produce L‐lactic acid, which could be a high additional value byproduct of ethanol production. The fermentation process of L‐lactic acid production by a newly isolated Enterococcus hawaiiensis CICIM‐CU B0114 is reported for the first time. RESULTS: The strain produced 56 g L^?1 of L‐lactic acid after cultivation for 48 h in optimized medium consisting of (g L^?1) 80 glucose, 10 peptone, 10 yeast extract, 1.5 Na₂HPO₄ and 0.2 MgSO₄. E. hawaiiensis CICIM‐CU B0114 was isolated and purified by subculture for growing and producing L‐lactic acid in distillery wastewater of very high gravity (VHG) from ethanol fermentation. L‐lactic acid fermentation was further studied with distillery wastewater substrate in 7 L and 15 L fermentors. The results showed that L‐lactic acid concentrations of 52 g L^?1 and 68 g L^?1 was achieved in 7 L and 15 L fermentors with the initial sugar concentrations of 67 g L^?1 and 87 g L^?1, respectively. CONCLUSION: The production of L‐lactic acid by the newly isolated E. hawaiiensis CICIM‐CU B0114 was carried out and the fermentation medium was optimized by orthogonal experimental design. This new strain holds the promise of L‐lactic acid production utilizing distillery wastewater from VHG ethanol fermentation. Copyright © 2010 Society of Chemical Industry     

Experimental evaluation of alternative fermentation media for L‐lactic acid production from apple pomace

BACKGROUND: A variety of nitrogen sources were tested for lactic acid production. Corn steep liquor is a low‐cost by‐product that could replace some of the expensive nutrients of the general lactobacilli media. This work deals with the optimisation of the composition of a low‐cost medium for lactic acid production from apple pomace by Lactobacillus rhamnosus CECT‐288. RESULTS: Corn steep liquor (CSL) and yeast extract (YE) were evaluated as nutrient sources for lactic acid production from apple pomace. In comparison with media containing CSL, experiments with YE led to higher volumetric productivities but lower lactic acid concentrations and product yields. The presence of YE increased the production of acetic acid. In media containing 0.2 g CSL g^?1 potential sugars, 29.5 g lactic acid L^?1 was obtained after 24 h, at good yield (35.5 g per 100 g dry apple pomace) and productivity (1.23 g L^?1 h^?1), with a lactic acid/acetic acid mass ratio of 98 g g^?1. CONCLUSION: The experimental results proved that CSL is a suitable nutrient source for lactic acid production from apple pomace, even though the volumetric productivity was lower than in experiments employing YE. Considered as a nutrient supplement, CSL presents advantages over YE not only in terms of cost but also in terms of product yield, final lactic acid concentration and lactic acid/acetic acid mass ratio. The utilisation of apple pomace as the raw material and CSL as the sole nutrient source (both cheap by‐products) enables the production of lactic acid by an economical, environmentally friendly process. Copyright © 2008 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     

Study of the bio‐production of carotenoids by Sporidiobolus salmonicolor (CBS 2636) using pre‐treated agro‐industrial substrates

BACKGROUND: The increasing industrial demand for carotenoids has aroused interest in their bio‐production, and the need to reduce production costs has encouraged the use of low cost industrial substrates, such as agro‐industrial residues. Thus the objective of this research was the bio‐production of carotenoids by Sporidiobolus salmonicolor using agro‐industrial substrates (corn steep liquor and sugarcane molasses), pre‐treated with acids (sulphuric and phosphoric). RESULTS: Bio‐production was carried out in an orbital shaker using a 10% (v/v) inoculum, incubation at 25 °C, and agitation at 180 rpm for 120 h in a non‐illuminated environment. The carotenoids were recovered using liquid N₂ combined with dimethylsulphoxide for cell rupture, and an acetone/methanol mixture (7:3 v/v) for extraction. CONCLUSION: The complete second‐order design allowed for optimisation of the carotenoid concentration obtained from industrial substrates pre‐treated with acids (sulphuric and phosphoric), obtaining a total carotenoid content of 541.5 µg L^?1 using 10 g L^?1 sugarcane molasses, 5 g L^?1 corn steep liquor and 5 g L^?1 yeast hydrolysate at 25 °C, with agitation at 180 rpm and an initial pH of 4.0. Copyright © 2008 Society of Chemical Industry     

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     

L(+)‐Lactic acid production using poly(vinyl alcohol)‐cryogel‐entrapped Rhizopus oryzae fungal cells

A new immobilized biocatalyst based on Rhizopus oryzae fungal cells entrapped in poly(vinyl alcohol)‐cryogel was evaluated in both the batch and semi‐batch processes of L (+)‐lactic acid (LA) production, when glucose, acid hydrolysates of starch or gelatinized potato starch were used as the main substrates. Under the batch conditions, the immobilized biocatalyst developed produced LA with yields of 94% and 78% from glucose and acid starch hydrolysates, respectively. Semi‐batch conditions enabled product yields of 52% and 45% to be obtained with the corresponding substrates. The highest process productivity (up to 173 g L^?1) was reached under semi‐batch conditions. Potato starch (5–70 g L^?1) was also transformed into lactic acid by immobilized R. oryzae. It was shown that long‐term operation of the immobilized biocatalyst (for at least 480 h) produced a low decrease in metabolic activity. Copyright © 2006 Society of Chemical Industry     

Lactic acid production from dining‐hall food waste by Lactobacillus plantarum using response surface methodology

BACKGROUND: Food waste generally has a high starch content and is rich in nutritional compounds, including lipids and proteins. It therefore represents a potential renewable resource. In this study, dining‐hall food waste was used as a substrate for lactic acid production, and response surface methodology was employed to optimise the fermentation conditions. RESULTS: Lactic acid biosynthesis was significantly affected by the interaction of protease and temperature. Protease, temperature and CaCO₃ had significant linear effects on lactic acid production, while α‐amylase and yeast extract had insignificant effects. The optimal conditions were found to be an α‐amylase activity of 13.86 U g^?1 dried food waste, a protease activity of 2.12 U g^?1 dried food waste, a temperature of 29.31 °C and a CaCO₃ concentration of 62.67 g L^?1, which resulted in a maximum lactic acid concentration of 98.51 g L^?1 (88.75% yield). An increase in inoculum size would be appropriate for accelerating the depletion of initial soluble carbohydrate to enhance the efficiency of α‐amylase in dining‐hall food waste fermentation. CONCLUSION: A suitable regression model for lactic acid production was developed based on the experimental results. Dining‐hall food waste was found to be a good substrate for lactic acid fermentation with high product yield and without nutrient supplementation. Copyright © 2008 Society of Chemical Industry     

Biotechnological production of lactic acid integrated with potato wastewater treatment by Rhizopus arrhizus

This paper describes a feasibility study of a for lactic acid production integrated with are treatment of wastewater from an industrial starch plant. Rhizopus oryzae two strains, Rhizopus arrhizus and Rhizopus oligosporus were tested with respect to their capability to carry out simultaneous saccharification and fermentation to lactic acid using potato wastewater. Rhizopus arrhizus DAR 36017 was identified as a suitable strain that demonstrated a high capacity for starch saccharification and lactic acid synthesis. The optimal conditions, in terms of pH, temperature and starch concentration, for lactic acid production were determined. The selected fungal strain grew well in a pH range from 3.0 to 7.0. The addition of CaCO₃10 g dm^?3 maintained the pH at 5.0–6.0 and significantly enhanced lactic acid production. Kinetic study revealed that almost complete starch saccharification and a lactic acid yield of 450g kg^?1 could be achieved in 20 h and 28 h cultivation, respectively. The maximum lactic acid production 21 g dm^?3 and mycelial biomass (1.7 g dm^?3) were obtained at 30 °C. Besides the multiple bioproducts, total removal of suspended solids and 90% reduction of COD were achieved in a single no‐aseptic operation. Copyright © 2003 Society of Chemical Industry     

Purification and enzymatic properties of the extracellular and constitutive chitosanase produced by Bacillus subtilis RKY3

BACKGROUND: Purification and enzymatic properties of a chitosanase from Bacillus subtilis RKY3 have been investigated to produce a chitooligosaccharide. The enzyme reported was extracellular and constitutive, which was purified by two sequential steps including ammonium sulfate precipitation and ion exchange chromatography. RESULTS: Sodium dodecyl sulfate‐polyacrylamide gel electrophoresis of the purified chitosanase revealed one single band corresponding to a molecular weight of around 24 kDa. The highest chitosanase activity was found to be at pH 6.0 and at 60 °C. Although the mercaptide forming agents such as Hg²⁺ (10 mmol L^?1) and p‐hydroxymercuribenzoic acid (1 mmol L^?1, 10 mmol L^?1) significantly or totally inhibited the enzyme activity, its activity was enhanced by the presence of 10 mmol L^?1 Mn²⁺. The enzyme showed activity for hydrolysis of soluble chitosan and glycol chitosan, but colloidal chitin, carboxymethyl cellulose, crystalline cellulose, and soluble starch were not hydrolyzed. The analysis of chitosan hydrolysis by thin‐layer chromatography and viscosity variation revealed that the purified enzyme should be endosplitting‐type chitosanase. CONCLUSION: The chitosanase produced by Bacillus subtilis RKY3 was a novel chitosanlytic enzyme with relatively low molecular weight, which is a versatile enzyme for chitosan hydrolysis because it could hydrolyze soluble chitosan into a biofunctional oligosaccharide at a high level. Copyright © 2011 Society of Chemical Industry     

Kinetics of glucoamylase hydrolysis of corn starch

Corn starch was hydrolysed by glucoamylase (EC 3.2.1.3, 1,4‐a‐D ‐glucan glucohydrolase) in a chemostat; 106 g L^?1 reducing sugar was produced at 40 °C after 46 h, whereas only 42 g L^?1 was produced at 30 °C. When corn starch concentration was increased from 64.6 g L^?1 to 161.5 g L^?1, the amount of reducing sugar produced increased with the initial substrate concentration. The studies show that there is a product concentration (p₀) above which product inhibition becomes apparent. A new model for starch hydrolysis considering the product inhibition point is presented. Copyright © 2006 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     

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     

Stabilized production of highly concentrated bioethanol from fermentation broths by Zymomonas mobilis by pervaporation using silicone rubber‐coated silicalite membranes

Since pervaporation performance of ethanol‐permselective silicalite membrane, which is an aluminum‐free hydrophobic zeolite, in the separation of fermentation broths by yeast are negatively affected by succinic acid, the potential of pervaporation using silicone rubber‐coated silicalite membranes of ethanol fermentation broths, not containing succinic acid, by Zymomonas mobilis was investigated for the reliable production of concentrated bioethanol. In the separation of fermentation broths, the pervaporation performance was influenced by nutrients used for the preparation of fermentation broths. In the separation of a broth prepared with yeast extract, pervaporation performance was greatly compromised by accumulation of a substance(s) having an ultraviolet absorption maximum at approximately 260 nm not only in total flux, but also in permeate ethanol concentration compared to the separation of binary ethanol/water mixtures. When supplying a prepared broth with corn steep liquor without the accumulation of a substance(s) having an ultraviolet absorption maximum at approximately 260 nm, the permeate ethanol concentration did not decrease. Treating the prepared broth with activated carbon was effective in restraining the decrease in total flux. Pervaporation performance is also deteriorated by the adsorption of lactic acid contained in corn steep liquor onto the silicalite crystals. In the separation of ternary mixtures of ethanol/water/lactic acid, accomplished by adjusting the ternary mixtures to pH > 5, more than 90% of the permeation flux in the separation of binary ethanol/water mixtures was obtained, and the permeate ethanol concentration was comparable to that obtained in the separation of binary mixtures. For stably performing pervaporation, it is important to prepare ethanol fermentation broths by Zymomonas mobilis in which lactic acid concentration is as low as possible. 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     

Fermentative production of mitomycins by Streptomyces caespitosus

Production of mitomycin antibiotics by Streptomyces caespitosus was increased by the utilisation of molasses (1.0%) treated with 0.01 % potassium ferrocyanide and corn steep liquor (0.9 %). Three different fermentation media were used for the fermentation production of mitomycins. Appropriate amounts of ammonium sulphate, soluble starch, ferrous sulphate, magnesium sulphate, potassium dihydrogen phosphate, sodium chloride and calcium carbonate increased the antibiotic yield when added to the fermentation medium containing molasses and corn steep liquor. The addition of some individual amino acids, vitamins and organic acids to a base fermentation medium did not improve or increase its performance.     

Lactic acid fermentation of food waste using integrated glucoamylase production

Commercial enzyme is usually needed for the bioconversion of organic waste or biomass. The overall cost could be reduced very significantly if enzyme production could be integrated with its application, avoiding unnecessary steps in enzyme production (such as concentration, recovery and transportation). This investigation attempted to integrate crude glucoamylase production with lactic acid fermentation of food waste. A maximum glucoamylase activity of 1850 U g^?1 was obtained with Aspergillus nigerduring solid‐state fermentation (SSF) of food waste, 14.8 times more than that obtained during submerged fermentation (SmF). The optimum pH for producing glucoamylase was 4.6, and glucoamylase retained 83.5% of peak activity at pH 3.0. Without any recovery treatment, the glucoamylase produced by SSF could be used directly for lactic acid fermentation of food waste. Lactic acid concentration reached 45.5 g L^?1 with the addition of the crude enzyme, 72% higher than the control. No side‐effects were caused by the viable A. niger in the crude enzyme. This work successfully integrated glucoamylase production with lactic acid fermentation. The enzyme produced by SSF of food waste had sufficient activity to be used directly without any treatment. The integrated process proposed in this study was very economical and may be helpful to other bioconversions. Copyright © 2008 Society of Chemical Industry     

High cell density fermentation via a metabolically engineered Escherichia coli for the enhanced production of succinic acid

BACKGROUND: Succinic acid is a valuable four‐carbon organic chemical with applications in many fields. It was found that cell mass was an important factor in succinic acid production by metabolically engineered Escherichia coli strains. In this work, high cell density fermentation was investigated for succinic acid production by a metabolically engineered strain SD121 with ldhA, pflB, ptsG mutation and heterogenous cyanobacterial ppc overexpression. RESULTS: Under two‐stage cultivation, the controlled DO feeding strategy during the aerobic growth phase facilitated biomass up to a dry cell weight of 19.6 g L^?1, and enhanced succinic acid production in the following anaerobic fermentation phase to a concentration of 116.2 g L^?1. A near theoretical maximum succinic acid yield of 1.73 mol mol^?1 glucose was achieved with an average productivity of 1.55 g L^?1 h^?1. CONCLUSION: The results indicated the potential advantage of high cell density fermentation for improvement of succinic acid production by E. coli. Copyright © 2010 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     

D‐Lactic acid production from waste cardboard

The effects caused by alkaline treatment on the susceptibility of waste cardboard to enzymatic hydrolysis have been studied. Optimised conditions leading to extensive saccharification of both cellulose (870 g kg^?1 conversion) and hemicelluloses (845 g kg^?1 conversion) were identified. Samples treated under selected operational conditions were employed for producing D ‐lactic acid by simultaneous saccharification and fermentation (SSF) in media containing cellulases, β‐glucosidase and Lactobacillus coryniformis ssp torquens cells. SSF fed‐batch experiments led to D ‐lactic acid concentrations up to 23.4 g dm^?3 at a product yield of 514 g lactic acid kg^?1 of potential glucose and a volumetric productivity of 0.48 g dm^?3 h^?1. Copyright © 2004 Society of Chemical Industry     

Downstream processing of 1,3‐propanediol fermentation broth

The downstream processing of 1,3‐propanediol fermentation broth using flocculation, reactive extraction, and reactive distillation was studied. Cellular debris and soluble protein in the broth were flocculated by combined use of chitosan and polyacrylamide at optimal concentrations of 150 ppm and 70 ppm, respectively; the soluble protein in the broth decreased to 0.06 g L^?1, and the recovery ratio of the supernatant liquor to broth was greater than 99%. 1,3‐Propanediol and other alcohols were extracted from the supernatant liquor by reacting with butyraldehyde. In a four‐stage countercurrent extraction with the volume ratio of the extraction solvent to the aqueous phase being 20:100, more than 99% 1,3‐propanediol acetal (2‐propyl‐1,3‐dioxane) and 2,3‐butanediol acetal (2‐propyl‐4,5‐dimethyl‐1,3‐dioxolane) were recovered from the aqueous phase; 35% of the glycerol acetals were recovered. The acetals produced were hydrolyzed in a reactive distillation column using the strongly acidic cation‐exchange resin as catalyst, the bottom product obtained was a mixture of 1,3‐propanediol (407 g L^?1), 2,3‐butanediol (252 g L^?1), glycerol (277 g L^?1), and glycerol acetals (146 g L^?1). Copyright © 2005 Society of Chemical Industry