Table of Contents Production of Maltodextrin and Glucose from Cassava Starch by Enzyme Hydrolysis using Locally Produced �-Amylase and Glucoamylase TERESITA M. ESPINO*, RICHARD D. TAMBALO, JACK DEODATO C. JACOB, FIDES Z. TAMBALO and PAUL ARJUNE OLAYRES National Institute of Molecular Biology and Biotechnology (BIOTECH), University of the Philippines Los Baños, College, Laguna, Philippines 4031 (*Corresponding author; e-mail: [email protected]) � � ���� �������� ��������� �������� ������� ����� ���������� ���� ��� ������������� ����� ����� ���� ���� ����������� ��� ������������� ���� ��������������������������������������������������������������������� ��� ���� ������� ����� ��������� ���� ���� ����������� ������ ����������� ��� ������������� ���� �������� ����� �������� ������� ������ ���� �������� ����������������������������������������������������������������������� ����� ������������ ������� ���������������� ��������� ��� ���� �������� ������� ����� ���� �������� ������ ���������� ���� �������� ���� ��� ��������� �� ����� ������� ������ ��� �������� ������� ���� ��������� ������ ����������������������������������������������������������������������� ��� ���� ����������������� ����������� ��� ���� ��������� ��������� ����� �������� ������� ��������� ����� ���� ���� ���� ��� ����������� ������ ������ ������ ��� ���� ����������� ���� ��������� �������� ��������� ����� �� ������ ��� ���� ������� ���� ��� �������������� ����� �� ��������� ���������� ��� ����������������������������������������������������������������� ��������������������������������������������������������������������� � ������������������������������������������������������������������ Introduction The Philippines, being a major agricultural country has an abundant supply of raw materials that can be utilized for the production of starches. Local sources of starch are grains (e.g. rice, corn, sorghum, wheat) and roots and tubers (sweet potato, cassava, potato, arrowroot). On a worldwide basis, cassava has now become the fourth most important source of calories after rice, sugarcane and maize. Cassava is highly productive which under favorable conditions yields up to 90 tons fresh roots per hectare which is equivalent to 27 tons of dry mass, 90% of which is pure starch. Under harsh environments, particularly where prolonged drought and poor soils are the major constraints for successful production of other staple food crops such as cereals, cassava crops produce reasonable yields [1]. This 949 949 characteristic has earned cassava the name 'famine fighter'. Often, cassava is the only crop people in the subtropics and tropics have to eat when other crops have died from drought. As a crop, cassava is one of the most efficient producers of starch, which constitutes about 85 percent of the storage root tissue dry-matter content. Annual production of cassava starch for industrial uses is about 800,000 tonnes, and originates mainly from Brazil (for the national market) and Thailand (for export to Japan and the EC) [2]. Starch is a high polymeric carbohydrate, with a molecular formula of (C6H10O5)n where n varies from a few hundred to over one million. Starch can be cooked as food or can be used as ingredient in foods. Moreover, it is used by the pharmaceutical, textile, paper and confectionery industries as binders, fillers and additives [3]. Starch can also be used as substrate for the production of high value products such as maltodextrin and glucose. Glucose, also called dextrose, is a moderately sweet sugar found in fruits and vegetables. It is mainly used in the baking, beverage (beer and malt liquor), canning, confectionery and dairy industries. It is also used for the production of intravenous fluids for medical and veterinary purposes [4]. Glucose can also be used for the production of ethanol and lactate, the raw material for biodegradable plastics [5]. Hydrolysis of starch into maltodextrin and glucose could be done either by acid hydrolysis, enzymatic hydrolysis or combination of both. Studies conducted showed that the use of microbial enzymes for starch hydrolysis is rapidly being utilized instead of acid hydrolysis [6]. The enzymes that are primarily involved in starch hydrolysis are amylases and glucoamylases. Considering that starch is universally available, an appropriate technology is urgently needed to produce high value products such as glucose and maltodextrin. Development of economically viable technologies will produce positive chain effect, which will benefit the farmers, food processors and finally consumers. Materials and Methods Materials Food grade cassava starch was purchased from the local market while the �-amylase and glucoamylase used were produced at BIOTECH, UP Los Baños using AM-1 bacterial isolate and the fungal culture, Aspergillus niger BIOTECH 3104, respectively. Glucose and maltodextrin standards were purchased from Sigma Chemical Co. Maltodextrin and glucose production Optimized conditions obtained by Espino et al. [7] were utilized for the laboratory scale production of maltodextrin and glucose from cassava starch using BIOTECH bacterial �–amylase and fungal glucoamylase. Production was carried out using a 30 L bioreactor. 950 950 High performance liquid chromatography (HPLC) analyses of samples Samples for HPLC were analyzed according to the method prescribed by Merck [8] using acetonitrile-water (70:30) as the mobile phase at 1 mL/min. A Lichrosper® 100 NH2 column 4�250 was used in the analysis. Glucose and maltodextrins (maltose, maltotriose, maltotetraose, maltopentaose, maltohexaose and maltoheptaose) were used as standards. All standards used were HPLC grade. Determination of physical and chemical properties of the produced glucose monohydrate Physical properties (melting point, solubility, odor and taste, color of solution and microscopic particle count test) and chemical properties (moisture content, pH, chloride content, specific rotation, sulfate level, acidity, presence of residue on ignition, presence of heavy metal expressed as lead, dextrin content, presence of unhydrolyzed starch, presence of 5-hydroxymethylfurfural and other related substances and dextrose level) of the BIOTECH produced glucose monohydrate from cassava starch were determined to ascertain the quality of the product and for comparison with the established USP standards. Toxicological evaluation of the glucose product The method of AOAC [9] was adapted for the preparation of water and oil extracts to be used in the chick embryo assay for toxicological evaluation. Fertile chicken eggs were purchased from the Institute of Animal Science (IAS), U.P. Los Baños. Total plate count Serial dilutions of the glucose product (DF 5, 10, 20 and 50) were pour-plated in Plate Count Agar (PCA) for the determination of total microbial count. Results and Discussion Maltodextrin and glucose production Optimized conditions obtained by Espino et al. [7] were utilized for the laboratory scale production of maltodextrin and glucose from cassava starch using BIOTECH bacterial �–amylase and fungal glucoamylase. Production was carried out using a 30 L 3.28 8.67 15.58 3.31 solvent 8.71 glucose maltose 15.67 3.31 (b) (a) Fig. 1. High Performance Liquid Chromatograms of standard (a) and BIOTECH-produced glucose derived from cassava starch (b). 951 951 bioreactor. ������� �� ������ ���� ����� ��������� ��� ���� ���������� ����������� ��� ���� �������� ������ ��������� ����� �������� ���� �������� ���������� ������� ��������� ���������������������������������������������������������������������������������� ������ ����� ���������� ����� ���� ����� ���������� ������ ��� ���� ����������� ������������� ��������� ������� ����� ���� �������� ������ ���������� ���� �������� ���� ��� ��������� �������������� ���� �� ��� �������� ������� ���� ��� ��������� ������ ����������������������������������������������������������������������������������� ������ ��� �������� ����� ������������ ������� ��������������� ���������� ����� ���� ������������������������������������������������������������������������� Table 1. Comparison of physical, chemical and microbiological properties of the BIOTECH-produced dextrose monohydrate from cassava starch with the United States Pharmacopoeia (USP) standards. Parameter BIOTECH dextrose monohydrate from cassava starch USP Requirements Identification test Passes Passes Odor Taste Color of solution Melting point, 0C Solubility, g/100 mL H2O Specific rotation, degrees Moisture content, %w/w Residue on ignition, %w/w None Sweetish Colorless 77.67 ± 0.44 None Sweetish Colorless 83.00 96.40 ± 0.94a 47.02b +52.33 ± 6.18 7.85 ± 0.75 0.021 ± 0.00 +52.6 to +53.2 7.5 to 9.5 �0.100 Acidity, as ppm CaCO3 Chloride, ppm 18.38 ± 3.19 8.20 ± 6.37 �5.00 �5.00 Sulfates, ppm Dextrins Soluble starch Lead, ppm pH Particulate matter, particles/mL: (25�m) (10 �m) 5-hydromethylfurfurals and related substances, Abs284nm Microbial limit, CFU/mL Possible toxins Assay for dextrose, %w/v �5 None None 0.56 3.92 ± 0.31 �5 None None <1.00 3.2 to 6.5 5.33 ± 1.78 9.17 ± 4.11 2 12 0.192 ± 0.02 �0.250 0 None 4.75 ± 0.29 0 None 5.00 952 952 953 ������ ��� Preliminary toxicological evaluation of the glucose product from cassava starch using chick embryo assay (AOAC, 1980)a. Number of eggs injectedb Number of live eggs Number of dead eggs % Survivalc 10 10 0 100 10 10 0 100 3 Water-soluble sample 10 9 1 90.0 4 Sterilized coconut oil (CONTROL 3) 10 9 1 90.0 5 Oil-soluble extract 10 9 1 90.0 Treatment Uninjected (CONTROL 1) 2 Sterilized distilled water (CONTROL 2) 1 a Average of two (2) trials. Incubated at 38�C for 14 days after injection. c % survival = number of live eggs X 100 number of eggs injected b Conclusion Two BIOTECH produced enzymes namely AM-1 bacterial �-amylase and AN fungal glucoamylase were used for the production of maltodextrin and glucose from cassava starch. A high 90-95% yield of glucose powder was obtained after lyophilization of the glucose monohydrate syrup. Results showed that majority of the physico-chemical properties of the dextrose produced from cassava starch complied with the USP set of standards. Total plate count of the unfiltered and filtered glucose solutions were <10 CFU/mL and 0, respectively. Results of the chick embryo assay showed that the glucose product is safe from oil soluble and water soluble toxins as shown by the high percent survival of 90-100% recorded for all the injected treatments. Acknowledgement This study was part of the project entitled, “High Value Products from Starches of Rootcrops by Enzyme Biotechnology” funded by the Agribusiness and Marketing Assistance Service (AMAS) Program through Region IV Department of Agriculture. The research on �–amylase was part of the project entitled, “Production of �–amylase for Food and Other Industrial Applications” funded by the Philippine Council for Industry and Energy Research and Development (PCIERD), DOST. 954 954 Table of Contents References 1. El-Sharkawy M.A. (1993). Drought-tolerant cassava for Africa, Asia, and Latin America. Bioscience 43: 441-451. 2. Woolfe, J. A. (1992). Sweet Potato: An Untapped Food Resource, 235 pp. Cambridge University Press. U.S.A. 3. Bujang, K., Adeni, D.S.A., and Jolhiri, P. (2000). Effects of starch concentration and pH on enzymic hydolysis of sago starch. Biotechnol. for Sust.Util. of Biol.Res. in the Trop. 14:32-35. 4. Dordick J.S. (1991). Biocatalysts for Industry, 243 pp. Plenum Press. New York and London. Chromcircle (Version 1.3). 2000. Merck KGaA, 65271 Darmstadt, Germany. 5. James, J. A. and Lee, B. H. (1997). Glucoamylases: Microbial sources, industrial application and molecular biology. A Review. J. Food Biochem. 21:1-52. 6. Coleman, G. and W.H. Elliott, W.H. (1962). Studies on �-amylase formation of Bacillus subtilis. Biochem. J. 83: 256-263. 7. Espino, T.M., Tambalo, R.D., Tambalo, F.Z., and Jacob, J.D.C. (2003). TERMINAL REPORT: High value products from starches of root crops by enzyme biotechnology. Department of Agriculture (DA)-Agribusiness and Marketing Assistance Service (AMAS) Program. Unpublished report. 8. Chromcircle (Version 1.3). (2000). Merck KGaA, 65271 Darmstadt, Germany. 9. Association of Official Analytical Chemists (AOAC). (1980). Official method of analysis of the Assoc. of Official Analytical Chemists. 13th ed. Horowitz (ed.), Washington, D.C., USA. 955 955
© Copyright 2026 Paperzz