Probiotic Enriched Noodles and Bread from Fermented Cassava Flour

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1 Probiotics in Sustainable Food Production: Current Status and Future Prospects - Probiotic Foods 136 Probiotic Enriched Noodles and Bread from Fermented Cassava Flour P.S. Geetha, Susheela A. Thirumaran and I. Maheswari Abstract--- Cassava (Manihot esculenta, Crantz), also called manioc, tapioca or yucca, is one of the most important food crops in the humid tropics and ranks as one of the main crops in the tropical countries. But cassava roots contain some cyanide in the form of cyanogenic glucosides, notably the linamarine, which can constitute a poison for the consumers when roots are processed improperly. The cassava root was cleaned, peeled and chipped and fermented for 48 hours using mixed culture like Saccharomyces cerevisae and two Lactobacillus Spp. Cassava flour, refined wheat flour and defatted soy flour were used in different combinations to produce instant cassava noodles and bread. Chemical (proximate, free fatty acid, peroxide value) and sensory properties of the instant cassava-refined wheat flour-defatted soy flour incorporated noodles were analyzed. The result obtained from the proximate analysis showed that increase in percentage of cassava in the noodle sample gave an increase in the carbohydrate, ash and fiber content respectively. There were significant differences (P<0.05) in the sensory attributes (colour, aroma, appearance, flavor, taste and texture) of the instant fermented cassava - refined wheat flour defatted soybean flour samples. New and alternative nutritious breads, can be produced from fermented cassava flour fortified with 20 per cent defatted soy flour, using locally available margarine and bakers yeast which increase the loaf volume by 25%.Margarine reduced the extent of starch gelatinization rendering them less gummy. The calcium, phosphorus and iron content of bread was increased when compare to control bread. The very important essential amino acids like lysine and metheonine were also comparatively high in fermented cassava flour and defatted soy flour incorporated bread. Organoleptic characteristics revealed the crust and crumb colour and crumb appearance of bread was highly acceptable. The bread was highly acceptable by the consumers and has good keeping qualities.a noodle produced from 100% wheat flour (control) was the most acceptable product by the panelists. The calcium, phosphorus and iron content of noodles was increased when compare to control. The very important essential amino acids like lysine and metheonine were also comparatively high in fermented cassava flour and defatted soy flour incorporated noodles. Cooking time of developed noodles from composite flour (15-18 min) was significantly (p<0.01) higher than cooking time of branded noodles (9.3 min). Mean overall organoleptic score of developed noodle from composite flour was in the range of highly acceptable criteria (20-25). By all means, 20% level of defatted soy flour blend incorporation was found to be acceptable. I. INTRODUCTION R OOT and tuber crops are second only to cereals in importance as a global source of food carbohydrate. Hence, they are aptly categorized as energy rich food crops due to their high starch content. Root and tuber crops are an important component in the diversification of agriculture to provide Food, Nutrition and Environmental Security. The tuber crops are especially significant in many developing countries more particularly in humid tropics where they are staple food to nearly million people. The process of fermentation has been found to bring about number of changes in the biochemical characteristics of the tubers. The disadvantage of lower nutritive value in cassava fermented products can easily be overcome by incorporation of the deficient nutrients before consumption. To overcome the protein and essential amino acids deficiency, the legume flours are incorporated. Defatted soy flour (DSF) is the most widely used soy ingredient in the baking industry, because soy flour can bind and hold large amount of water P.S. Geetha, Assistant Professor, Dept. of FRM, Home Science College & Research Institute, TNAU, Madurai. geethafsn@gmail.com Susheela A.Thirumaran, Former Dean, Home Science College & Research Institute, TNAU, Madurai. I. Maheswari, Project Fellow, Dept. of FRM, Home Science College & Research Institute, TNAU, Madurai.

2 Probiotics in Sustainable Food Production: Current Status and Future Prospects - Probiotic Foods 137 through cooking process which is estimated to be more than three times of their weight in water (Porter and Skarra, 1999), It is the prime objective of this paper to anchor this research on the following specific objectives: 1. To standardize processes for cassava fermentation for sour flours. 2. To studying their nutritional and anti-nutritional characteristics 3. To develop the value added products from cassava sour flour II. MATERIALS AND METHODS Investigation were undertaken to study the Processing of value added fermented cassava base pulse flour incorporated products and study their quality in the Department of Food Science and Nutrition, Home Science College and Research Institute, Madurai, India. Fully matured, good quality cassava tubers were procured from the fields of Athur, Salem District in Tamil Nadu. The other items like refined wheat flour (RWF), DSF, black gram flour, vanaspathi, citric acid, ice cream ready-mix powder, corn flour, milk cream, oil and other minor groceries were purchased from the local market. The packaging materials, viz., brown paper bag, gunny bags, polyethylene pack of 100 and 200 gauges thickness (20 x 15 cm.), and plastic container (PET) (250 ml cap) were used for packing the products for conducting the storage study. The equipment used were Brabender farinograph, (OMG, Duisberg, West Germany), Brabender amylograph (Duisberg, West Germany). Extruder (Model Dolly) Bacterial Culture Selected microorganisms of Lactobacillus cellobiosus, Lactobacillus plantarum and Streptococcus lactis in lypolised form were obtained from the National Institute of Dairy Technology Kanpur and used. Processing of Flour Processing of Fermented Sour Cassava Flour (FSCF) The dried chips were powdered and passed through a 30-mesh sieve to get FSCF. Sieve Analysis of Flour The cassava flour, refined wheat flour, defatted soy flour were sieved through a series of sieves viz., Bs. 60, 80, 100, 120 and 200. The percentage of flour, which passed through each sieve, was calculated. Formula for Extruded Products Based on the studies conducted earlier by Thirumaran Susheela (1993), a formula for the extruded products was developed. Cassava flour and Refined Wheat Flour were mixed in three ratios namely 1:3, 1:1

3 Probiotics in Sustainable Food Production: Current Status and Future Prospects - Probiotic Foods 138 and 3:1. To the selected base of 1:3 and 1:1 DSF was incorporated at 15, 20, 25 and 30 per cent levels. Thus the formula for noodles was developed. To the first base the DSF was incorporated at 15, 20, 25 and 30 per cent levels and was reported. Processing of Noodles Fermented Cassava Based Bread (FSCF) Along with FSCF and RWF, DSF was incorporated (15-30 percent) while preparing bread, bun and biscuits. The proportion of ingredients used (formula development) for each treatment. Determination of Farinograph Characteristic of the Flour The procedure suggested by Venkateswara Rao and Haridas Rao (1993) was followed. Farinograph records the resistance offered by a dough to the mixing blades during a prolonged and relative gentile mixing at a constant temperature by transmitting the resistance to the dynometer. The dynometer, in turn is connected to a lever and scale system and to a pen which traces a curve called farinogram. Amylograph characteristics of the flour The procedure suggested by Venkateswara Rao and Haridas RAo (1993) was adopted. Determination of Fat and Water Absorption Fat and water absorption capacity of FSCF blends were measured by the method described by Sharma (1985) with slight modifications. Physical characteristics of the processed cassava products such as Sieve analysis, Gelatinization time and temperature, Rheologcal characteristics of flour were also done. Breaking Strength of Noodles To study the breaking strength of noodles, a simple technique was developed. A noodle strip of 6.00 cm length was taken and its ends were tied horizontally to the supporting stand at about 15cm above the ground level. A known weight (5, 10 and 20g) was dropped on the tied strip at different heights. When the noodle strip breaks, the breaking strength of the noodles was recorded as Y cm. It was analysed periodically. Storage Studies Two numbers of bread and noodles were packed in each packaging material, viz., low density polyethylene bags and high density polyethylene bags and kept in storage for a period of one month at room temperature and their chemical constituents were analysed. Hundred gram of noodles of base-i and Base-II were packed separately in Low Density PolyEthylene (LDPE) and High Density PolyEthylene (HDPE) bags and stored for a period of six months at room temperature (32-36 o C, per cent RH) to study their storage stability.

4 Probiotics in Sustainable Food Production: Current Status and Future Prospects - Probiotic Foods 139 Cooking Quality of Extruded Products Cooking quality of the stored noodles was tested. The parameters such as cooking time, cooked volume, cooked weight and water absorption were studied by following the method described by Singh et al. (1980). III. RESULTS AND DISCUSSION Farinograph studies in base-i and base-ii, 25:75, 50:50 and per cent incorporation of DSF blends were evaluated. Water absorption index of the control (T0) was observed as 63.0 ml. in base-i, T1 and T5 values noted were 85 and 95 ml each. In base-ii, both T6 and T10 had 90 and 100 ml each (Fig.1 to 4) Fig. 1 Fig. 2 Fig. 3 Fig. 4 Fermentation influenced the functional properties exhibited by the soybean flour, particularly in swelling, water and oil absorption. The control flour with a crude protein content of 10 per cent showed a water absorption capacity of 13 per cent. While the cassava flour with a crude protein content of about two per cent showed a water absorption value of 21 per cent. It is evident that though protein had been associated with water binding properties in flours, the non-protein components play an important role. The presence of starch, which is hydrophilic polymer, in the cassava may have contributed appreciably to the high water absorption capacity exhibited by the flour despite its low protein content. The bakery product bread was packed in LDPE and HDPE packaging materials and stored at room temperature to study the storage behaviours, viz., chemical changes, microbial load and consumer acceptability. The statistically analysed data are presented and discussed. PHYSICAL Characteristics of Bread and Noodles The bulk density of control bread was g / cc. For various treatments it ranged between to g/cc for bread. The data clearly indicated that addition of DSF resulted in the increase in weight and reduction in volume in bread. The cooking quality was observed for water absorption, cooked volume, cooking time, solid loss and rehydration characteristics. Initial samples of control noodles showed a value of ml, which changed to 318.oo and ml for the samples irrespective of packaging materials at the end of storage. The mean values for DSF incorporated processed noodles ranged from to ml for base-i and to ml for base-ii. The cooked volume of the noodles increased as the level of noodles was 213 whereas for base-i it ranged from 228 to 290 and fro base-ii 220 to 283 ml. The mean cooking time ranged from to for base-ii to minutes. The solid loss of the noodles increased when the DSF level were increased. Maximum solid loss was up to g per cent. The rehydration ratio also increased with the increase in DSF substitution. The mean value of base-i and base-ii was ranged from 9.70, to and to ml respectively. All the cooking qualities showed highly significant difference between treatments and packaging materials during storage period.

5 Probiotics in Sustainable Food Production: Current Status and Future Prospects - Probiotic Foods 140 Plate 2: Bread Plate 1: Noodles Chemical Analysis The moisture content of bread samples were gradually found to increase when the level of DSF was increased. The moisture content of control bread and bun was and per cent while it ranged between and per cent in base-i, and and per cent in base II in bread. The moisture content of the bun ranged between and per cent in base I and and per cent in base- II. A gradual increase in the moisture content was observed in all the noodle samples packed in different packaging materials stored at room temperature throughout the storage period. It ranged between , to and to for control base I and base-ii respectively. The protein contents of the control bread was g per cent. The values for base-i and base-ii ranged from to and 7.03 to g percent. It was noticed that at 30 per cent level of DSF, the protein contents of bread increased by about per cent when compared to control. The mean protein content of the noodles showed increased values with increased DSF incorporation levels. The 25 and 30 per cent incorporations showed minimal differences and there was no significant difference. The T4 and T5 in base -I blends and T9 and T10 in base-ii blends were on par with each other and the values ranged from to and to g per cent respectively. The lowest level of and per cent was exhibited by 25:75 and 50:50 plain cassava and refined wheat flour processed noodles. Fig.5: Changes in Protein (g%) Content of Bread during Storage The total sugar content of the bread was for control; to for base-i and to g per cent for base-ii. The data on total sugar content of freshly prepared noodles showed a steady increase and had mean values of for control, to for base-i and to g per cent for base-ii. A highly significant difference in the total sugar content of noodles was observed in respect of treatments and storage period. The initial reducing sugar content of bread for control sample was 5.40 per cent. For the base-i bread, the values ranged from 4.50 to 7.51 g per cent and base-ii, the values ranged from 8.51 to g per cent. As the difference in increase of reducing sugar was very meager, it would not have any influence in baking quality. The initially no change was observed in the reducing sugar content of the noodles. A slight increase was noticed from 60 th day of noodles storage packed in HDPE and LDPE packs. The reducing sugar content of noodles ranged from 6.96 to for base-i and 6.58 to g per cent for base-ii. The data revealed a highly significant difference in the reducing sugar content of noodles between treatments, packaging materials and period of storage.

6 Probiotics in Sustainable Food Production: Current Status and Future Prospects - Probiotic Foods 141 The calcium content of bread was increased gradually at each incremental addition of DSF. The calcium content of control bread was mg per cent. In treatments, the values ranged from to and to g per cent in base-i and base-ii respectively. Phosphorus content of bread also showed an increasing trend. The iron content slightly ranged high when compared to control for bread. The control bread showed values of 8.50, while the same in base-i was 5.90 to and in base-ii 4.00 to mg per cent. The calcium content in general increased after storage. It might be due to the breakdown of complex compounds and decrease of moisture content. It ranged from 44 to 118 for base-i and 57 to 142 mg per cent for base-ii noodles. The minimum was exhibited by plain RWF noodles (24 mg per cent) and the maximum by DSF incorporated noodles. A range of to for base-i and to mg per cent for base-ii were observed. The control RWF noodles exhibited a value of mg per cent. Phosphoorus content also increased except in T 1 and T6. This could also be attributed as to the calcium. The minimum iron content was exhibited by RWF noodles whereas the maximum (6.28 mg per cent) by 30 per cent DSF incorporated noodles. Similarly the value ranged between 2.06 to 4.16 for base-i and 3.24 to 6.28 mg per cent for base-ii. Under shelf condition of the mineral content of noodles remained unchanged in some of the treatment whereas it was high and slightly reduced in others. The lysine content of bread was mg per cent in control, whereas the same for bread in base-i and base-ii respectively ranged from to and to The methionine content of bread showed the values of mg per cent for control, to for base-i and to mg per cent for base-ii respectively. The amino acids content showed that the lysine and methionine contents respectively ranged from and for control samples, to and to mg/g N of lysine for base-i and base-ii and to and to mg/g N of methionine for base-i and base-ii processed noodles. The lysine content increased and methionine content decreased compared to control. Microbial Load Fig.6 Fig.7 Fig. 6 & 7: Amino Acid Pattern in Noodles (Methionine and lysine) The initial fungal and yeast count for bread was analysed by using 10-4 dilution. The counts ranged between 00 and 03 according to treatment irrespective of the packaging materials. Presence of yeast population was noted in all the stored noodle samples. The maximum and minimum count of fungi was found to be 4 x 10-4 and 1 x 10-4 cells /g which were within the tolerable limits. Organoleptic Characteristics The bread supplemented up to 20 per cent DSF was well liked by 93 per cent of the respondents. The overall acceptability value of noodles was found to be 8.56 for base-i and 8.30 for base-ii processed noodles. The shelf life of DSF incorporated bread 4 days and all the treatment of the noodles can be stored up to 6 months.

7 Probiotics in Sustainable Food Production: Current Status and Future Prospects - Probiotic Foods 142 IV. CONCLUSION From this study, it is concluded that cassava fermentation is an advantageous proposition in reducing the toxicity due to cyanide, improving the functional properties of resultant starch and imparting flavor and taste. The cost of protein rich legume incorporated noodles was lesser compared to control noodles and can be advocated for commercial productions. The present study also established that the noodles available in the market also found to be much higher than the different indigenous types of extruded noodles. RECOMMENDATIONS Supplementation of fermented cassava with protein rich food can definitely be used to upgrade cassava as a Value Added Food Product (VAFP), pave way for consumption to a much larger extent and thereby Nutritional Security can be sustained among under privileged people especially in the under developed countries. This can be achieved through identification of more number of protein rich locally available energy rich native crops. REFERENCES [1] Acquistucei, R The Maillard reaction in pasta, Preliminary classification by multivariate techniques. Cer. Chem. 20(7): [2] Akingbala, J.O., G.B.Ogunticen, and O.A.Sobanke, Physico-chemical properties of yam flour substituted with soy flour. Plant food and human nutrition, 48: [3] Goyle, A and M.Mathur, Sensory evaluation and acceptability of breads prepared from refined wheat and soy bean flours. The Indian Journal of Nutritional dietetics, 36: [4] Samuel, A., Studies on the development of cereal millets pulse based bakery products. M.Sc. Thesis (unpub), Home Science College, Agricultural College & Research Institute, TamilNadu Agricultural University, Madurai. [5] Dixit, H., S.S.Shukla, S.Shrivastava and Y.K.Sharma, Studies on soy dhal flour blends and the shelf life of soy dhal. Ind. J. Nutr. Dietet. 23(10): [6] Falcone and Phillips, Development of extruded products based with locally available cereal and pulse flour. Food and Nutrition Bulletin, 17(20): [7] Indrani, D.G., D.Savithri and G.Venkateshwara Rao, Effect of defatted soy flour on the quality of bun. FSTA., 32(10): 670. [8] Mestress, C., P.Collanna and A.Baleon Characteristics of starch network within rice flours noodles and mung bean starch vermicelli. Starche/Starke, 68(9): [9] Mponda, G.T., Cassava and soybean flours and resting time effects on sensory and physical characteristics of reconstituted mionda. J. Sci. Food and Agriculture, 96(2): [10] Porter, M.A and L.L.Skarra, Reducing cost through inclusion of soy flour in breads. Cereal Food World 44(9): [11] Roulet, M.C., Solubilisation of sugar beet pulp cell wall poly sacchrides by extrusion cooking. Lebnasm. Wiss Unite Technology. (Paris) 24: [12] Sahare, G.M., P.M.Kolicha and S.S.Kadam, Effect of incorporation of defatted soybean meal on organoleptic nutritional properties of bakery products. Beverage and Food World, 26(6): 36.