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Starch Update 2011: The 6 th International Conference on Starch Technology P-STARCH-4 Functionality benchmarking of underutilized starches with cassava starch Sirithorn Lertphanich 1, Rungtiva Wansuksri 2, Thierry Tran 3, Guillaume Da 4, Luong Hong Nga 5, Kuakoon Piyachomkwan 2 and Klanarong Sriroth 1 1Department of biotechnology, Faculty of Agro-Industry, Kasetsart University, Bangkok, Thailand 2Cassava and Starch Technology Research Unit, National Center for Genetic Engineering and Biotechnology (BIOTEC), Bangkok, Thailand 3Centre de Coopération Internationale en Recherche Agronomique Pour le Développement (CIRAD), Dpt. Persyst, UMR Qualisud, Montpellier, France 4 CERTES, Université Paris-Est Créteil, 61 avenue du Général de Gaulle, 94000 Créteil, France 5 Hanoi University of Science and Technology (HUST), IBFT, Hanoi, Vietnam Abstract Plant organs represent a highly diverse source of reserve starch but many crops remain underutilized for agro-industries. In this study, structural and functional properties of starches from various storage organs including from tubers, i.e.yam bean ( Pachyrhizus erosus ), from corms, i.e.taro ( Colocasia esculenta ) and ensete ( Enseteventricosum ), from fruits, i.e. water caltrop ( Trapa natans ) and from grains (legumes), i.e. chickpea (Cicer arietinum ) and mungbean ( Vigna radiata ) were characterized and compared with cassava starch. All extracted starches were pure (protein and ash < 0.25% dwb). Taro starch had the smallest granules (2 µm) while ensete granules were the largest (42 µm). Only ensete starch had the B-type polymorph, while mungbean and chickpea starches were C-type and the rest were A- type. Yam bean, water caltrop and ensete starches had amylose contents of 14.15, 19.20 and 20.85%, respectively, similar to cassava starch (17.44%), while taro starch was lower (7.91%) and mungbean and chickpea starches were higher (28.47% and 35.59%. With regard to functional properties, different gelatinization, pasting and rheological properties were observed. Based on peak viscosity as determined by Rapid Visco Analyzer (9.2% starch dsb), three groups of starches were classified; high (cassava and mungbean; 319 and 373 RVU), medium (yam bean, water caltrop and ensete with 222-281 RVU) and low (taro and chickpea with 197 and 176 RVU, respectively). Interestingly, only water caltrop, mungbean and chickpea had a positive value of setback from peak, implying high tendency of gelation. The dynamic rheological analysis also revealed high storage modulus (G ′) values upon cooling for those three starches, while cassava, yam bean and taro had low G ′ values. Taro had the highest susceptibility to α-amylase hydrolysis, while ensete starch granules had the lowest. These results demonstrate various starches with diverse properties, comparatively to cassava starch and can potentially find value-added uses as food ingredients. Keywords: Cassava, ensete, water caltrop, chickpea, yam bean, taro, mungbean, paste, rheology 1. Introduction Starch is the second most abundant polysaccharide after cellulose as an energy reserve in many photosynthesizing plants. It is found in different storage organs including seeds (corn, wheat, rice, barley, sorghum), tubers (potato, yam), roots (cassava), fruits (mango, banana), stems (sago palm), rhizomes (canna), corms (taro).These are employed as an important staple food in many regions and are also used for starch production. Extracted starches are extensively employed in a wide range of food and non-food application, functioning as thickeners, binders, emulsifiers, texture modifiers, gelling agent, film forming agent and so 180 Starch Update 2011: The 6 th International Conference on Starch Technology on. Nowadays, major commercial starches are produced from seeds, i.e. corn, wheat and rice, from tubers, i.e. potato and from roots, i.e. cassava. In tropical areas where high plant biodiversity is recognized, there are still many diversified starch-reserved plants being underutilized. Starches from different botanical origins are likely to have diversified structural and physico-chemical properties. In this study, properties of starches from various underutilized crops grown in tropical regions were characterized and compared with cassava starch. 2. Materials and Methods 2.1. Materials Cassava (Manihot esculenta ), Mungbean (Vigna radiat a),Taro (Colocasia esculenta ), Yam bean (Pachyrhizus erosus ), Chickpea(Cicer arietinum ) were collected from local markets in Thailand. Water Caltrop (Trapa natans ) was taken from Vietnam. Ensete(Enseteventricosum ) was obtained from a local market in Ethiopia. 2.2 Methods Starch extraction and composition analysis Starch was extracted by grinding peeled samples with water and passing through a 170-mesh sieve. Starch slurry was then collected and settled. After starch sedimentation, water was decanted and starch cake was rewashed twice before drying. Starch purity was determined by an enzyme method (AACC, 1995). If the purity was less than 90%, the sample was further washed with sodium hydroxide solution. The content of protein, fat, fiber and ash were also quantified according to the AOAC methods (AOAC, 1990). Structural properties: Granule morphology and amylose content Granule morphology of starch samples was observed under JEOL scanning electron microscopy (JSM-5310, England) at 10-KV acceleration. The X-ray patterns of starches were obtained with Cu-K∝ radiation using a diffractometer (Joel X-ray diffractometer JDX-3080, Joel, Japan), scanning from 5 ° to 40 ° 2 θ at 0.02 ° 2 θ/min. Iodine affinity measurement by an automatic amperometric Titrator (835 Titrando,Metrohm, Herisau, Switzerland) was performed to quantify the amylose content, according to the method of Takeda & Hizukuri (1987). Viscosity properties Viscosity properties of the starch suspension (9.2 % dwb) were evaluated using a Rapid Visco Analyzer (RVA4, Newport Scientific, Australia). The starch suspentsion was heated from 50 to 95 °C at a rate of 9 °C/min and held at 95 °C for 3 min . After that, starch paste was cooled down to 50 °C at the same rate. The viscosity of starch paste, including peak viscosity (P), viscosity at 95 °C (H), viscosity at 50 °C (C) and gelatinization temperature were recorded. The breakdown (P-H) and setback from peak (C-P) were also reported. Rheological measurements Rheological properties of the starch suspensions (20%w/w) were determined on a rotational Physica MCR 300 rheometer (Physica Messtechnik GmbH, Stuttgart, Germany) with a plate and plate geometry sensor (50 mm diameter, and 1mm gap). A thin layer of light paraffin oil was added to prevent evaporative loss. Temperature ramp sweep test of the samples was performed from 25 to 95 °C, and from 95 to 25 °C at a rate of 1.5 °C/min. Parameters including storage modulus (G ′), loss modulus (G ′′) and loss tangent (tan δ = G ′/G ′′) were monitored at frequency of 1 Hz and a strain of 0.5 % (thelinear viscoelasticity domain). 181 Starch Update 2011: The 6 th International Conference on Starch Technology In vitro digestibility In vitro starch digestibility was analyzed according to the method of Miao et al. (2009). The starch sample (200 mg) suspended and prewarmed in phosphate buffer (15 ml, 0.2 mol/l, pH 5.2,37 oC) were incubated with porcine pancreatic α-amylase (290 U/ml) and amyloglucosidase (15 U/ml)with shaking (150 rpm). Aliquots of hydrolysed solution (0.5 ml) were collected at various reaction times. The glucose content of centrifuged hydrolyzate was determined using the glucose oxidase-peroxidase assay kit (Megazyme, Wicklow, Ireland). 3. Results and Discussion 3.1 Starch composition analysis Extracted starches prepared in this study were quite pure as indicated by very low quantities of impurities, i.e. protein, lipid and ash (in total not greater than 0.5% dwb). The starch contents by enzymatic assay of all extracted samples were greater than 95% dwb. 3.2 Granule morphology and amylose content Starch granule morphology was discrete and species-specific (Table1). Taro starch granules were polyglonal with the smallestsize of 2 µm, followed by yam bean starch with the spherical granules of 8.2 µm in average. Among all starches, ensete starch had the largestangular granules of 42 µm. Granule morphology of mungbean and chickpea starches was quite similar in shape, i.e. oval and round and size (17.7 and 21.0 µm, respectively). The X-ray diffraction displayed an A-type polymorph for cassava, taro, yam bean and water caltrop while ensete starch possessed a B-type crystal polymorph. Similar to typical pea starches, both mungbean and chickpea starches were C-type. The amylose contents of extracted starches were different, ranging from 7.9% in taro starch to 35.6%for chickpea starch (Table 1). 3.3 Pasting properties During heating, insoluble starch granules can absorb water, swell to much larger sizes with some granule disruption and glucan leaching. Accordingly, the starch suspension becomes to paste with changes its flowability. The abrupt increase in paste viscosity of starches from different sources occurred at different temperatures and reached to different maximum viscosity (Table 2). Paste viscosity of ensete starch having the largest granules was developed at the lowest temperature (69.80 °C) while taro starch with the smallest granules had rapid viscosity change at a very high temperature (82.35 °C) with low peak viscosity (197 RVU). The highest peak viscosity of cooked paste was observed in mungbean starch (373 RVU), followed by cassava starch (319 RVU) with high breakdown