Solid-state fermentation using wheat bran to produce glucose syrup and functional cereal bars.

Wheat bran is one of the most abundant by-products from grain milling, which can be used as substrate for solid-state fermentation (SSF) to obtain enzymes able to convert this agro-industrial waste into glucose syrup, which in turn can be applied for the production of different food products. The present study aimed to determine centesimal composition of wheat bran, obtain enzymatic extract that converts wheat bran into wheat glucose syrup (WGS), produce rice flakes cereal bars (RFCB), and evaluate their nutritional composition and the presence of functional compounds, as well as their antioxidant potential. Determination of centesimal composition of wheat bran demonstrated its nutritional potential. Enzymatic extract was obtained and it converted wheat bran into WGS, which were applied to rice flakes producing RFCB. These cereal bars proved to be a source of dietary fiber (1.8 g) and soluble protein (7.2 g) while RCFB produced with corn glucose syrup did not present these nutritional components. In addition, RFCB produced with WGS showed polyphenolic compounds, among them flavonoids, which exhibited antioxidant activity by DPPH and ABTS radical scavenging (47.46% and 711.89 µM Trolox Equivalent/g, respectively), and iron ion reduction (71.70 µM Trolox equivalent/g). Final product showed a decrease in caloric value and sodium content. Therefore, the present study showed that the bioprocess of SSF yields a nutritional, ecological, and functional food product, which might be of great interest for food industry, adding nutritional and functional value to a well-stablished product.

Toxicity and Anti-Inflammatory Effects of Agave sisalana Extract Derived from Agroindustrial Residue.

BACKGROUND: In several countries, the leaf juice of Agave sisalana (also known as sisal) is widely used topically, especially as an antiseptic, and orally for the treatment of different pathologies. However, in Brazil, which is the largest producer of Agave sisalana, its residue, which represents the majority of its weight, has been thrown away. For this reason, the determination of the pharmacological and toxicological potentials of sisal residue and its possible therapeutic use is seen as a way to contribute to the sustainable development and social promotion of the largest producer of sisal in Brazil, the interior of Bahia State, which is among the poorest areas in the country. Given the scarcity of available scientific studies on the pharmacological and toxicological properties of sisal residue juice, this study aimed to promote the acid hydrolysis of this juice to potentiate the anti-inflammatory effect already described in the literature. Furthermore, it aimed to evaluate the toxicological profile of the hydrolyzed extract (EAH) and to determine its acute toxicity, as well as its side effects on the reproductive aspects of rats. METHOD: The anti-inflammatory effect of EAH was evaluated in vitro using the induction of hemolysis by hypotonic solution and in vivo in rats using the carrageenan-induced paw edema test and the xylene-induced ear edema test. The acute toxicity, resulting from a single-dose administration, was investigated for some manifestation of toxic symptoms related to motor control and consciousness in rats. At a concentration of 100 mg/kg, by repeated doses, the reproductive toxicity effects of EAH in rats were assessed. RESULTS: In vitro anti-inflammatory activity was positive using the human red blood cell membrane stabilization method. In both in vivo tests used to assess the anti-inflammatory activity, EAH (at three doses) significantly inhibited edema when compared to the control group. At a dose of 50 mg/kg, EAH exhibited a greater effect than indomethacin, a nonsteroidal anti-inflammatory drug with known activity. In vivo toxicological studies have shown that EAH does not present toxic effects when administered orally in a single dose, up to 1000 mg/kg. Finally, EAH promoted a gonadotoxic effect and increased the embryonic mortality rate after implantation. CONCLUSIONS: It is suggested that the anti-edematogenic effect of the acid hydrolysis extract from sisal juice is due to the high concentration of steroidal sapogenins. Therefore, this extract can be considered a potential new anti-inflammatory or even an important sapogenin source for the development of steroidal glucocorticoids. However, further studies are needed to elucidate the chemical composition of sisal juice. Regarding toxicology studies, EAH did not show cytotoxic and clastogenic potentials, but it presented a powerful reproductive toxic effect in rats.

Fibrolytic enzymes improving in vitro rumen degradability of tropical forages.

During specific times of the year, especially dry seasons, tropical forages typically have poor nutritional value due to high contents of neutral and acid detergent fibres, and low crude protein in their composition, which may reduce productivity of ruminant livestock production and lead to increased enteric methane (CH4 ) emissions per unit of generated product in forage-bases systems. In order to increase fibre degradability and the efficiency of energy utilisation from low-quality forages, exogenous fibrolytic enzymes have been studied. In this assay, we evaluated the effects of increasing dose levels of fibrolytic enzymes extract (FEE) produced by Trichoderma reesei on in vitro rumen organic matter degradability, fermentation parameters, total gas and CH4 production of tropical forages. Forage samples were analysed for their bromatological composition, and enzyme activity from FEE was performed for xylanase and endoglucanase. The in vitro gas production technique was used in a 5 × 3 factorial arrangement with five FEE dose levels (0, 5, 50, 500 and 5,000 µl) and three substrates (Cynodon spp., Panicum maximum and Cenchrus ciliaris L.). The highest dose level of FEE increased degradability, total gas and CH4 production in all substrates (p < .05). Butyrate concentration also increased while acetate:propionate ratio and pH decreased with the addition of FEE (p < .05). These results indicated that the use of fibrolytic enzymes can be a reliable strategy to improve degradability of low-quality forages, contributing to the sustainability and intensification of livestock production in tropical countries.

Ethanol from rice byproduct using amylases secreted by Rhizopus microsporus var. oligosporus. Enzyme partial purification and characterization.

A three-stage bioethanol bioprocess was developed. Firstly, amylases were obtained from Rhizopus microsporus var. oligosporus using wheat bran in solid-state fermentation. Secondly, amylases hydrolyzed a rice byproduct to make a glucose-rich solution, and this sugar was finally metabolized by Saccharomyces cerevisiae to produce bioethanol. Besides, the secreted enzymes were also partially purified and characterized. The amylase activity (AA) in the crude extract was 358 U/g substrate, and the partially purified enzyme showed the best activity in the 4.0-5.5 pH range. A wide pH stability range (3.5-8.5) was confirmed. The amylase was thermostable up to 60 °C. The ion Mn+2 (10 mM) improved by 60% the AA. There was a 54.9% yield in the conversion of rice residues into reducing sugars in 10 h. The glucose-rich solution was undergone fermentation by S. cerevisiae and showed high ethanol efficiency, 95.8% of the theoretical value. These results suggested a promising technology for bioethanol production.

Use of ultraviolet-visible spectrophotometry associated with artificial neural networks as an alternative for determining the water quality index.

The water quality index (WQI) is an important tool for water resource management and planning. However, it has major disadvantages: the generation of chemical waste, is costly, and time-consuming. In order to overcome these drawbacks, we propose to simplify this index determination by replacing traditional analytical methods with ultraviolet-visible (UV-Vis) spectrophotometry associated with artificial neural network (ANN). A total of 100 water samples were collected from two rivers located in Assis, SP, Brazil and calculated the WQI by the conventional method. UV-Vis spectral analyses between 190 and 800 nm were also performed for each sample followed by principal component analysis (PCA) aiming to reduce the number of variables. The scores of the principal components were used as input to calibrate a three-layer feed-forward neural network. Output layer was defined by the WQI values. The modeling efforts showed that the optimal ANN architecture was 19-16-1, trainlm as training function, root-mean-square error (RMSE) 0.5813, determination coefficient between observed and predicted values (R2) of 0.9857 (p < 0.0001), and mean absolute percentage error (MAPE) of 0.57% ± 0.51%. The implications of this work's results open up the possibility to use a portable UV-Vis spectrophotometer connected to a computer to predict the WQI in places where there is no required infrastructure to determine the WQI by the conventional method as well as to monitor water body's in real time.

Analysis of antioxidant, cytotoxic and mutagenic potential of Agave sisalana Perrine extracts using Vero cells, human lymphocytes and mice polychromatic erythrocytes.

Brazilian Northeast is the world's largest producer of Agave sisalana Perrine for the supply of the sisal fiber. About 95% of plant biomass, which comprise the mucilage and sisal juice, is considered a waste residual is discarded in the soil. However, the sisal juice is rich in steroidal saponins, which exhibits different pharmacological properties. Despite this, natural products are not necessarily safe. Based on this, this study analyzed the antioxidant, cytotoxic and mutagenic potential of three extracts derived from acid hydrolysis (AHAS), dried precipitate (DPAS) and hexanic of A. sisalana (HAS). These analyses were performed by in vitro and in vivo methods, using Vero cells, human lymphocytes and mice. Results showed that AHAS 50 and 100 can be considered a useful antineoplastic candidate due to their antioxidant and cytotoxic activity, with no genotoxic/clastogenic potential in Vero cells and mice. Although the comet assay in human lymphocytes has showed that the AHAS 25, AHAS 50 and AHAS 100 can lead to DNA breaks, these extracts did not promote DNA damages in mice bone marrow. Considering the different mutagenic responses obtained with the different methods employed, this study suggest that the metabolizing pathways can produce by-products harmful to health. For this reason, it is mandatory to analyze the mutagenic potential by both in vitro and in vivo techniques, using cells derived from different species and origins.

The potential of tailoring the conditions of steam explosion to produce xylo-oligosaccharides from sugarcane bagasse.

In this study, the potential of the steam explosion (SE) method to produce high levels XOS from sugarcane bagasse, a xylan-rich hemicellulosic feedstock, was assessed. The effect of different operating conditions on XOS production yield and selectivity were investigated using a mini-pilot scale SE unit. The results show that even under a non-optimized condition (190 °C, 5 min and 0.5% H2SO4 as catalyst), SE led to about 40% xylan recovery as XOS, which was comparable to the well-known, multi-step, enzymatic production of XOS from alkaline-extracted xylan, and other commonly employed chemical methods. In addition, the XOS-rich hydrolysate from SE constituted of greater diversity in the degree of polymerization, which has been shown to be desirable for prebiotic application.

Analysis of antioxidant, cytotoxic and mutagenic potential of Agave sisalana Perrine extracts using Vero cells, human lymphocytes and mice polychromatic erythrocytes

Brazilian Northeast is the world's largest producer of Agave sisalana Perrine for the supply of the sisal fiber. About 95% of plant biomass, which comprise the mucilage and sisal juice, is considered a waste residual is discarded in the soil. However, the sisal juice is rich in steroidal saponins, which exhibits different pharmacological properties. Despite this, natural products are not necessarily safe. Based on this, this study analyzed the antioxidant, cytotoxic and mutagenic potential of three extracts derived from acid hydrolysis (AHAS), dried precipitate (DPAS) and hexanic of A. sisalana (HAS). These analyses were performed by in vitro and in vivo methods, using Vero cells, human lymphocytes and mice. Results showed that AHAS 50 and 100 can be considered a useful antineoplastic candidate due to their antioxidant and cytotoxic activity, with no genotoxic/clastogenic potential in Vero cells and mice. Although the comet assay in human lymphocytes has showed that the AHAS 25, AHAS 50 and AHAS 100 can lead to DNA breaks, these extracts did not promote DNA damages in mice bone marrow. Considering the different mutagenic responses obtained with the different methods employed, this study suggest that the metabolizing pathways can produce by-products harmful to health. For this reason, it is mandatory to analyze the mutagenic potential by both in vitro and in vivo techniques, using cells derived from different species and origins.

Analysis of antioxidant, cytotoxic and mutagenic potential of Agave sisalana Perrine extracts using Vero cells, human lymphocytes and mice polychromatic erythrocytes

Brazilian Northeast is the world's largest producer of Agave sisalana Perrine for the supply of the sisal fiber. About 95% of plant biomass, which comprise the mucilage and sisal juice, is considered a waste residual is discarded in the soil. However, the sisal juice is rich in steroidal saponins, which exhibits different pharmacological properties. Despite this, natural products are not necessarily safe. Based on this, this study analyzed the antioxidant, cytotoxic and mutagenic potential of three extracts derived from acid hydrolysis (AHAS), dried precipitate (DPAS) and hexanic of A. sisalana (HAS). These analyses were performed by in vitro and in vivo methods, using Vero cells, human lymphocytes and mice. Results showed that AHAS 50 and 100 can be considered a useful antineoplastic candidate due to their antioxidant and cytotoxic activity, with no genotoxic/clastogenic potential in Vero cells and mice. Although the comet assay in human lymphocytes has showed that the AHAS 25, AHAS 50 and AHAS 100 can lead to DNA breaks, these extracts did not promote DNA damages in mice bone marrow. Considering the different mutagenic responses obtained with the different methods employed, this study suggest that the metabolizing pathways can produce by-products harmful to health. For this reason, it is mandatory to analyze the mutagenic potential by both in vitro and in vivo techniques, using cells derived from different species and origins.

Lactic acid production from submerged fermentation of broken rice using undefined mixed culture.

The present work aimed to characterize and optimize the submerged fermentation of broken rice for lactic acid (LA) production using undefined mixed culture from dewatered activated sludge. A microorganism with amylolytic activity, which also produces LA, Lactobacillus amylovorus, was used as a control to assess the extent of mixed culture on LA yield. Three level full factorial designs were performed to optimize and define the influence of fermentation temperature (20-50 °C), gelatinization time (30-60 min) and broken rice concentration in culture medium (40-80 g L-1) on LA production in pure and undefined mixed culture. LA production in mixed culture (9.76 g L-1) increased in sixfold respect to pure culture in optimal assessed experimental conditions. The optimal conditions for maximizing LA yield in mixed culture bioprocess were 31 °C temperature, 45 min gelatinization time and 79 g L-1 broken rice concentration in culture medium. This study demonstrated the positive effect of undefined mixed culture from dewatered activated sludge to produce LA from culture medium formulated with broken rice. In addition, this work establishes the basis for an efficient and low-cost bioprocess to manufacture LA from this booming agro-industrial by-product.

Chemical input reduction in the arabinoxylan and lignocellulose alkaline extraction and xylooligosaccharides production.

Lignocellulosic material breakdown by hydrolysis is an important step to open new perspectives for bioenergy and special foods production like prebiotic xylooligosaccharides. Improvement of lignocellulose and arabinoxylan alkaline extraction from sugarcane bagasse and enzymatic hydrolysis were performed. Treatments 1 (10% KOH at 70°C), 3 (5% KOH at 121°C) and ZD method (24% KOH at 35°C) showed solid lignocellulose recovery of respectively 75.2%, 74.2% and 73%. A range of 24.8-27% extracted material with high arabinoxylan content (72.1-76.3%) was obtained with these treatments. Treatment 1 and 3 exhibited great KOH reduction in the method reaction, 54.1% and 76.2%, respectively. Likewise, in treatment 3 there was a decrease in ethanol consumption (40.9%) when compared to ZD method. The extracted arabinoxylan showed susceptibility to enzymatic hydrolysis with high solid loading (7%) since Trichoderma reesei xylanases were advantageous for xylose production (54.9%), while Aspergillus fumigatus xylanases achieved better XOS production (27.1%).

Properties of films obtained from biopolymers of different origins for skin lesions therapy

In this study, the effects of the origin of xanthan used, in combination with chitosan, to prepare films for the treatment of skin lesions were evaluated. The characteristics of the films obtained with xanthan commercially available for the food industry sector and xanthan originated from a fermentation process conducted in a pilot plant were compared. Results showed that the source did not strongly interfere in many of the properties of the films, such as the mechanical properties, cytotoxicity to L929 cells, absorption of simulated body fluid and culture medium, stability in water and saline solution. Hence, even though the properties of biopolymers of different sources might vary, the films prepared with two distinct types of xanthan gum could be considered as potentially safe and similar in terms of relevant characteristics considering the aimed application.

Immobilization of Papain on Chitin and Chitosan and Recycling of Soluble Enzyme for Deflocculation of Saccharomyces cerevisiae from Bioethanol Distilleries.

Yeast flocculation (Saccharomyces cerevisiae) is one of the most important problems in fuel ethanol production. Yeast flocculation causes operational difficulties and increase in the ethanol cost. Proteolytic enzymes can solve this problem since it does not depend on these changes. The recycling of soluble papain and the immobilization of this enzyme on chitin or chitosan were studied. Some cross-linking agents were evaluated in the action of proteolytic activity of papain. The glutaraldehyde (0.1-10% w·v(-1)), polyethyleneimine (0.5% v·v(-1)), and tripolyphosphate (1-10% w·v(-1)) inactivated the enzyme in this range, respectively. Glutaraldehyde inhibited all treatments of papain immobilization. The chitosan cross-linked with TPP in 5 h of reaction showed the yield of active immobilized enzyme of 15.7% and 6.07% in chitosan treated with 0.1% PEI. Although these immobilizations have been possible, these levels have not been enough to cause deflocculation of yeast cells. Free enzyme was efficient for yeast deflocculation in dosages of 3 to 4 g·L(-1). Recycling of soluble papain by centrifugation was effective for 14 cycles with yeast suspension in time perfectly compatible to industrial conditions. The reuse of proteases applied after yeast suspension by additional yeast centrifugation could be an alternative to cost reduction of these enzymes.

Combining xanthan and chitosan membranes to multipotent mesenchymal stromal cells as bioactive dressings for dermo-epidermal wounds.

The association between tridimensional scaffolds to cells of interest has provided excellent perspectives for obtaining viable complex tissues in vitro, such as skin, resulting in impressive advances in the field of tissue engineering applied to regenerative therapies. The use of multipotent mesenchymal stromal cells in the treatment of dermo-epidermal wounds is particularly promising due to several relevant properties of these cells, such as high capacity of proliferation in culture, potential of differentiation in multiple skin cell types, important paracrine and immunomodulatory effects, among others. Membranes of chitosan complexed with xanthan may be potentially useful as scaffolds for multipotent mesenchymal stromal cells, given that they present suitable physico-chemical characteristics and have adequate tridimensional structure for the adhesion, growth, and maintenance of cell function. Therefore, the purpose of this work was to assess the applicability of bioactive dressings associating dense and porous chitosan-xanthan membranes to multipotent mesenchymal stromal cells for the treatment of skin wounds. The membranes showed to be non-mutagenic and allowed efficient adhesion and proliferation of the mesenchymal stromal cells in vitro. In vivo assays performed with mesenchymal stromal cells grown on the surface of the dense membranes showed acceleration of wound healing in Wistar rats, thus indicating that the use of this cell-scaffold association for tissue engineering purposes is feasible and attractive.

Chemical inhibition of the contaminant Lactobacillus fermentum from distilleries producing fuel bioethanol

The purpose of this study was to determine the Minimum Inhibitory Concentration (MIC) of pure or mixed chemicals for Saccharomyces cerevisiae and Lactobacillus fermentum in the samples isolated from distilleries with serious bacterial contamination problems. The biocides, which showed the best results were: 3,4,4' trichlorocarbanilide (TCC), tested at pH 4.0 (MIC = 3.12 mg/l), TCC with benzethonium chloride (CBe) at pH 6.0 (MIC = 3.12 mg/l) and TCC mixed with benzalkonium chloride (CBa) at pH 6.0 (MIC = 1.53 mg /l). If CBa was used in sugar cane milling in 1:1 ratio with TCC, a 8 times reduction of CBa was possible. This formulation also should be tested in fermentation steps since it was more difficult for the bacterium to develop resistance to biocide. There was no inhibition of S. cerevisiae and there were only antibiotics as an option to bacterial control of fuel ethanol fermentation by S. cerevisiae.

Applications of the hexanic fraction of Agave sisalana Perrine ex Engelm (Asparagaceae): control of inflammation and pain screening.

The present study evaluated the anti-inflammatory and analgesic properties of Agave sisalana Perrine in classic models of inflammation and pain. The hexanic fraction of A. sisalana (HFAS) was obtained by acid hydrolysis followed by hexanic reflux. Anti-inflammatory properties were examined in three acute mouse models (xylene ear oedema, hind paw oedema and pleurisy) and a chronic mouse model (granuloma cotton pellet). The antinociceptive potential was evaluated in chemical (acetic-acid) and thermal (tail-flick and hot-plate test) models of pain. When given orally, HFAS (5, 10, 25 and 50 mg/kg) reduced ear oedema (p < 0.0001; 52%, 71%, 62% and 42%, respectively). HFAS also reduced hind paw oedema at doses of 10 mg/kg and 25 mg/kg (p < 0.05; 42% and 58%, respectively) and pleurisy at doses of 10 mg/kg and 25 mg/kg (41% and 50%, respectively). In a chronic model, HFAS reduced inflammation by 46% and 58% at doses of 10 mg/kg and 25 mg/kg, respectively. Moreover, this fraction showed analgesic properties against the abdominal writhing in an acetic acid model (at doses of 5-25 mg/kg) with inhibitory rates of 24%, 54% and 48%. The HFAS also showed an increased latency time in the hot-plate (23% and 28%) and tail-flick tests (61% and 66%) for the 25 mg/kg and 50 mg/kg doses, respectively. These results suggest that HFAS has anti-inflammatory and analgesic properties.

Applications of the hexanic fraction of Agave sisalana Perrine ex Engelm (Asparagaceae): control of inflammation and pain screening

The present study evaluated the anti-inflammatory and analgesic properties of Agave sisalana Perrine in classic models of inflammation and pain. The hexanic fraction of A. sisalana (HFAS) was obtained by acid hydrolysis followed by hexanic reflux. Anti-inflammatory properties were examined in three acute mouse models (xylene ear oedema, hind paw oedema and pleurisy) and a chronic mouse model (granuloma cotton pellet). The antinociceptive potential was evaluated in chemical (acetic-acid) and thermal (tail-flick and hot-plate test) models of pain. When given orally, HFAS (5, 10, 25 and 50 mg/kg) reduced ear oedema (p < 0.0001; 52%, 71%, 62% and 42%, respectively). HFAS also reduced hind paw oedema at doses of 10 mg/kg and 25 mg/kg (p < 0.05; 42% and 58%, respectively) and pleurisy at doses of 10 mg/kg and 25 mg/kg (41% and 50%, respectively). In a chronic model, HFAS reduced inflammation by 46% and 58% at doses of 10 mg/kg and 25 mg/kg, respectively. Moreover, this fraction showed analgesic properties against the abdominal writhing in an acetic acid model (at doses of 5-25 mg/kg) with inhibitory rates of 24%, 54% and 48%. The HFAS also showed an increased latency time in the hot-plate (23% and 28%) and tail-flick tests (61% and 66%) for the 25 mg/kg and 50 mg/kg doses, respectively. These results suggest that HFAS has anti-inflammatory and analgesic properties.

Improvement in RNA extraction from S. cerevisie by optimization in the autolysis and NH3 hydrolysis

The optimization of autolysis of Saccharomyces cerevisiae from brewery was studied aiming at the maximum ribonucleic acid extraction and yeast extract production. The best conditions for yeast autolysis was 55.2ºC, pH= 5.1 and 9.8 percent NaCl for 24h of processing, without the NH3 use. In these conditions, the RNA yield was 89.7 percent, resulting in 51.3 percent of dehydrated yeast extract with 57.9 percent protein. The use of 12.2 percent NH3 at 60ºC after autolysis (8h) and plasmolysis (8h) was not viable due to the reduction in the RNA yield from 89.7to78.4 percent. On the other hand, the thermal shock at 60ºC for 15 minutes prior to autolysis provided an increase in the yield from 89.7 to 91.4 percent. The autolysis, including NaCl plasmolysis in the optimized conditions was efficient, economic and with short time, thus usable for industrial purpose to obtain more valuable products such as yeast extract enriched in RNA and/or protein, for different applications.

New Culture Medium to Xanthan Production by Xanthomonas campestris pv. campestris.

Xanthan is a important biopolymer for commercial purpose and it is produced in two stages by Xanthomonas campestris. In the first one, the bacterium is cultivated in the complex medium enriched in nitrogen and the biomass produced is used as inoculum for the next stage in which the gum is produced in another medium. In this work a new medium for the first stage is proposed in place of currently used YM medium. Different formulated growth media were studied and the correspondent biomass produced was analysed as inoculum for the second stage. The inoculum and gum were produced by batch process in shaker at 27°C in pH 6.0 and at 30°C in pH 7.0, respectively. The gum was precipitated with ethanol (3:1 v/v). The dryed biomass and xathan gum produced were determined by drying in oven at 105 and 40°C, respectively. The viscosity of the fermentation broth and 1% gum solution in water were determined in Brookfield viscometer. The formulated medium presented the increase in gum production (30%), broth (136%) and 1% gum solution viscosity (60%) compared to YM, besides the inferior cost. The results showed the importance of the quality of the inoculum from the first stage of the culture which influenced on the gum viscosity in the second stage.

Sugarcane molasses and yeast powder used in the Fructooligosaccharides production by Aspergillus japonicus-FCL 119T and Aspergillus niger ATCC 20611.

Different concentrations of sucrose (3-25% w/v) and peptone (2-5% w/v) were studied in the formulation of media during the cultivation of Aspergillus japonicus-FCL 119T and Aspergillus niger ATCC 20611. Moreover, cane molasses (3.5-17.5% w/v total sugar) and yeast powder (1.5-5% w/v) were used as alternative nutrients for both strains' cultivation. These media were formulated for analysis of cellular growth, beta-Fructosyltransferase and Fructooligosaccharides (FOS) production. Transfructosylating activity (U ( t )) and FOS production were analyzed by HPLC. The highest enzyme production by both the strains was 3% (w/v) sucrose and 3% (w/v) peptone, or 3.5% (w/v) total sugars present in cane molasses and 1.5% (w/v) yeast powder. Cane molasses and yeast powder were as good as sucrose and peptone in the enzyme and FOS (around 60% w/w) production by studied strains.