Starch Hydrolysis Research Articles

Removal of proteins and lipids affects structure, in vitro digestion and physicochemical properties of rice flour modified by heat-moisture treatment.

The objective of this experiment was to investigate the role of endogenous proteins and lipids in the structural and physicochemical properties of starch in heat-moisture treatment (HMT) rice flour and to reveal their effect on starch digestibility under heat. The findings indicate that, in the absence of endogenous proteins and lipids acting as a physical barrier, especially proteins, the interaction between rice flour and endogenous proteins and lipids diminished. This reduction led to fewer starch-protein inclusion complexes and starch-lipid complexes, altering the granule aggregation structure of rice flour. It resulted in a decrease in particle size, an increase in agglomeration between starch granules, and more surface cracking on rice granules. Under HMT conditions with a moisture content of 30%, slight gelatinization of the starch granules occurred, contributing to an increased starch hydrolysis rate. In addition, the elevated thermal energy effect of HMT enhanced interactions between starch molecular chains. These resulted in a decrease in crystallinity, short-range ordering, and the content of double-helix structure within starch granules. These structural transformations led to higher pasting temperatures, improved hot and cold paste stability, and a decrease in peak viscosity, breakdown, setback, and enthalpy of pasting of the starch granules. The combined analysis of microstructure, physicochemical properties, and in vitro digestion characteristics has enabled us to further enhance our understanding of the interaction mechanisms between endogenous proteins, lipids, and starches during HMT. © 2024 Society of Chemical Industry.

The pseudoenzyme β-amylase9 from Arabidopsis binds to and enhances the activity of α-amylase3: A possible mechanism to promote stress-induced starch degradation.

Starch accumulation in plant tissues provides an important carbon source at night and for regrowth after periods of dormancy and in times of stress. Both ɑ- and β-amylases (AMYs and BAMs, respectively) catalyze starch hydrolysis, but their functional roles are unclear. Moreover, the presence of catalytically inactive amylases that show starch excess phenotypes when deleted presents an interesting series of questions on how starch degradation is regulated. Plants lacking one of these catalytically inactive β-amylases, BAM9, were shown to have enhanced starch accumulation when combined with mutations in BAM1 and BAM3, the primary starch degrading BAMs in response to stress and at night, respectively. Importantly, BAM9 has been reported to be transcriptionally induced by stress through activation of SnRK1. Using yeast two-hybrid experiments, we identified the plastid-localized AMY3 as a potential interaction partner for BAM9. We found that BAM9 interacted with AMY3 in vitro and that BAM9 enhances AMY3 activity 3-fold. Modeling of the AMY3-BAM9 complex revealed a previously undescribed N-terminal structural feature in AMY3 that we call the alpha-alpha hairpin that could serve as a potential interaction site. Additionally, AMY3 lacking the alpha-alpha hairpin is unaffected by BAM9. Structural analysis of AMY3 showed that it can form a homodimer in solution and that BAM9 appears to replace one of the AMY3 monomers to form a heterodimer. Collectively these data suggest that BAM9 is a pseudoamylase that activates AMY3 in response to cellular stress, possibly facilitating starch degradation to provide an additional energy source for stress recovery.

Production of gluconic acid from the washed rice waste water using Au/MgO catalyst – A sustainable route

The majority of the waste produced by the food and agriculture industries is abundant in proteins, carbohydrates, and fats, which can be utilized effectively in other food products or industrial products. Especially, washed rice water (WRW) contains a significant quantity of starch that has been discarded without being utilized properly. In the present investigation, we have successfully upgraded washed rice water into the industrially important intermediate, i.e., gluconic acid, using an Au/MgO catalyst in a single pot reaction. The upgrading strategy was developed in three consecutive phases using two different model reactions: (1) glucose to gluconic acid, (2) hydrolysis of starch into glucose, followed by the oxidation reaction. The results showed that almost 60% gluconic acid was achieved at room temperature with atmospheric pressure. The present investigation highlighted that hydrolysis, followed by oxidation reaction is the most promising route for upgrading WRW to gluconic acid.

Potentiality of Beneficial Microbe Bacillus siamensis GP-P8 for the Suppression of Anthracnose Pathogens and Pepper Plant Growth Promotion.

This study was carried out to screen the antifungal activity against Colletotrichum acutatum, Colletotrichum dematium, and Colletotrichum coccodes. Bacterial isolate GP-P8 from pepper soil was found to be effective against the tested pathogens with an average inhibition rate of 70.7% in in vitro dual culture assays. 16S rRNA gene sequencing analysis result showed that the effective bacterial isolate as Bacillus siamensis. Biochemical characterization of GP-P8 was also performed. According to the results, protease and cellulose, siderophore production, phosphate solubilization, starch hydrolysis, and indole-3-acetic acid production were shown by the GP-P8. Using specific primers, genes involved in the production of antibiotics, such as iturin, fengycin, difficidin, bacilysin, bacillibactin, surfactin, macrolactin, and bacillaene were also detected in B. siamensis GP-P8. Identification and analysis of volatile organic compounds through solid phase microextraction/gas chromatography-mass spectrometry (SPME/GC-MS) revealed that acetoin and 2,3-butanediol were produced by isolate GP-P8. In vivo tests showed that GP-P8 significantly reduced the anthracnose disease caused by C. acutatum, and enhanced the growth of pepper plant. Reverse transcription polymerase chain reaction analysis of pepper fruits revealed that GP-P8 treated pepper plants showed increased expression of immune genes such as CaPR1, CaPR4, CaNPR1, CaMAPK4, CaJA2, and CaERF53. These results strongly suggest that GP-P8 could be a promising biocontrol agent against pepper anthracnose disease and possibly a pepper plant growth-promoting agent.

A new route to the synthesis of 2-hydrazolyl-4-thiazolidinone hybrids, evaluation of α-glucosidase inhibitory activity and molecular modeling insights

One of the multifactorial worldwide health syndromes is diabetes mellitus which is increasing at a disturbing rate. The inhibition of α-glucosidase, an enzyme that catalyzes starch hydrolysis in the intestine, is one helpful therapeutic approach for controlling hyperglycemia related to type-2 diabetes. To discover α-glucosidase inhibitors, some 2-hydrazolyl-4-thiazolidinone hybrids (3a-e) were synthesized from new one-pot reaction procedures. Next, their chemical structures were confirmed by 1H NMR, 13C NMR, and FT-IR spectra, and elemental analysis technique. Then, the α-glucosidase inhibitory activity of the titled compounds was evaluated. Among them, derivatives 3b and 3c revealed the highest activity against α-glucosidase compared to acarbose as a drug. Enzyme kinetic studies of the most active derivative (3b) indicated a competitive inhibition. Finally, molecular modeling studies were accomplished to describe vital interactions of the most potent compounds (3b and 3c) with the α-glucosidase enzyme.

Prevalence and characterization of Xanthomonas axonopodis pv. cyamopsidis causing bacterial blight of clusterbean in the semi-arid region of North Gujarat

The bacterial blight caused by Xanthomonas axonopodis pv. cyamopsidis is the most widespread and destructive disease of clusterbean. The present study aimed to survey the major clusterbean growing areas, expanding in five districts i.e., Banaskantha, Patan, Sabarkantha, Mehsana and Aravali of North Gujarat. The study found that bacterial blight disease is prevalent in Idar, Ambaji, Danta, Sanali, Khedbramha, Shamalaji, Patan, Bhiloda, Visnagar, Deesa and Amirgadh. The highest mean disease incidence (28.44%) was recorded in Idar, Sabarkantha. Based on the gram straining it was observed that the test bacterium was gram-negative and rod-shaped. Further, the biochemical tests, such as starch hydrolysis, gelatin liquefaction, hydrogen sulphide production, catalase, and acid production from different sugars viz., sucrose, maltose, and dextrose showed a positive reaction, except, acid production from mannitol was negative. Based on the morphological, biochemical, and pathogenic characteristics, the pathogen was identified as Xanthomonas axonopodis pv. cyamopsidis.

Characterization of raw starch hydrolysing and detergent-compatible amylase from newly isolated Bacillus subtilis from bakery chimney

Amylases are highly needful product in food, beverage, and detergent industry. With the objective of isolating α-amylases producing strains, samples were collected from local bakery chimneys and kitchens, Chennai, Tamil Nadu, India. The potential α-amylase producers were isolated and screened by Starch-Iodine plate assay method. The optimized parameters such as 1.5% of starch concentration in the media with 7.2-pH at 35 °C for 36 h enhanced the production of α-amylase. The strain showing high potential, identified by 16s rRNA sequencing was found to be Bacillus subtilis. The Bacillus subtilis strain produced extracellular α-amylase with activity of 1330.14 U/mL. Further partial purification by ammonium sulphate precipitation followed by dialysis and column chromatography enhanced the specific activity to 1695.306 U/mg. The molecular weight of enzymes was found to be around 36 kDa. The enzyme stability and activity characterization revealed that the enzyme was thermotolerant which was able to retain more than 70% activity at 70 °C, 80 °C and 90 °C. The purified enzyme was also found to be Ca2+ independent. The hydrolysis of raw starch from rice, wheat and corn flour using the purified α-amylase from Bacillus subtilis showed significant activity of 72.8%, 76.1% and 65.2% with 4 h incubation respectively. The potentiality of the enzyme as an additive with commercial detergent was also determined to be effective. This study favourably attracts the attention towards industrial applications.

Effect of different pre-treatment on acrylamide content, nutrition value, starch digestibility and anthocyanin bioaccessibility of purple sweet potato (Ipomoea batata) deep-fried chips

Veggie chips have gained popularity in the European market. These are considered healthier than potato chips by consumers. However, few works evaluate their nutritional and digestibility. The current work aimed to evaluate the effect of four pre-frying treatments (soaking, blanching, pulsed electric field (PEF) and PEF + blanching combination (PEFB)) on the chemical composition, anthocyanins, acrylamide, and digestive behavior (starch hydrolysis and anthocyanins bioaccessibility) of purple sweet potato deep-fried chips. In total 15 independent batches were made, three for each studied treatment (also a control without pretreatment was developed). The studied pretreatments impacted on fat and starch content, especially blanching and PEFB, which caused an increase in fat absorption and break starch, generating maltodextrins. Nineteen anthocyanins were detected, mainly cyanidin and peonidin derivatives, but a drastic loss was observed in blanched, PEF-treated and PEF-B-Treated chips. Acrylamide values ranged from 504.11 to 6350.0- μg/kg, with the highest values reported by untreated chips and the lowest by PEF-B-treated chips (p < 0.05). The anthocyanin's bioaccessibility ranged between 66.57 and 92.88%, with soaked chips that showed the highest values.

Proteus mirabilis and Klebsiella variicola associated with onion bacterial streak and bulb rot

The bulb onion (Allium cepa) is second only to tomatoes in value of vegetable crops cultivated worldwide. Water-soaked lesions, bleaching and blight on leaves and slippery skin, sour skin, soft rot near the neck and centre rot on onion bulbs were observed in surveys of the onion fields, storage as well as fruit and vegetable markets of northwestern provinces of Iran in 2020. Thirty-four samples were collected from onion fields of East Azerbaijan, West Azerbaijan and Zanjan provinces and the central storage of Bonab city. Fifty bacterial strains were isolated from symptomatic samples. Pathogenicity of the strains was confirmed by inoculating on onion bulb cultivars Azarshahr and Ghermez Anari. Twenty-eight strains were able to induce soft rot on the inoculated sites of cut onion bulbs. Onion bulbs turned dark brown at lesion sites and became odorous over time. All pathogenic strains were Gram-negative, facultative aerobic and catalase positive. Strains mainly varied in starch and aesculin hydrolysis, potato rot and hypersensitive reaction. Pathogenic strains separated into four groups based on phenotypic assays. In the phylogenetic tree based on concatenated sequences of gyrB and infB genes, the strains were divided into four clusters containing the type strains of Enterobacter ludwigii, Klebsiella variicola, Proteus mirabilis and Pantoea agglomerans, confirming their identity as belonging to these four species. This is the first report of P. mirabilis as a plant pathogen and of K. variicola as a causal agent of onion bulb rot. Moreover, this is the first report of E. ludwigii and P. agglomerans associated with onion bulb rot in Iran.

Cutting-edge progress in green technologies for resistant starch type 3 and type 5 preparation: An updated review

Resistant starch (RS) is a dietary fiber that resists starch hydrolysis in the small intestine, and is fermented in the colon by microorganisms. RS not only has a broad range of benefits in the food and non-food industries but also has a significance impact on health promotion and prevention of non-communicable diseases. RS types 3 and 5 have been the focus of research from an environment-friendly perspective. RS3 is normally formed by recrystallization after physical modification, whereas RS5 is obtained by the complexation of starch and fatty acids through the thermomechanical methods. This review provides updates and approaches to RS3 and RS5 preparations that promote RS content based on green technologies. This information will be useful for future research on RS development and for identifying preparation methods for functional food.

Effect of exogenous abscisic acid on improving low-phosphorus (P) response in rice (Oryza sativa L.)

Phosphorus (P) deficiency is a major constraint to rice productivity worldwide. Exogenous application of abscisic acid (ABA) has not been reported to improve rice response toward P-deficiency. Present study analyzed the biochemical response in the shoots and roots of two rice cultivars, CSR10 (low-P tolerant) and Pusa44 (low-P susceptible) at 36, 72 and 216 h after normal P (NP) (10 ppm) (control), low P (LP) (1 ppm), and LP + ABA given to 12-d old seedlings. Results found that biomass and P-uptake reduced in Pusa44 compared to CSR10 under LP but improved significantly under LP + ABA. P-fractionation was utilized efficiently by CSR10 under LP while LP + ABA improved P-fractionation in both cultivars. LP + ABA improved acid phosphatases in both cultivars. Hexose and sucrose levels were depleted while starch hydrolyzed poorly in Pusa44 compared to CSR10 under LP while LP + ABA enhanced starch hydrolysis and levels of sugars in both cultivars. Starch increased at a late stage in the roots of CSR10 under LP while Pusa44 improved it under LP + ABA. Antioxidant capacity was low while H2O2 was high at early stage in Pusa44 compared to CSR10 under LP. LP + ABA improved antioxidant capacity and reduced H2O2 in both cultivars. Results concluded that ABA is a versatile regulator of not only P-acquisition/P-utilization but also carbohydrates and antioxidant activities under P-deficiency, and exogenous ABA improved low-P tolerance in rice.

New Derivatives of Modified Starch for Food Technology.

The food industry extensively uses chemically modified starches and their hydrolysates, which is mainly due to their emulsification ability. Therefore, it becomes inevitable to develop new starch derivatives, including modified starch hydrolysates, and effective preparation methods to meet the increasing demands of producers, consumers, and technology. This study comprehensively researches the physical, chemical, and functional properties (such as the water-binding capacity, swelling power, solubility, and fat absorption capacity) of chemically modified biopolymers and their enzymatic hydrolysis products. We utilized oxidized and acetylated potato and waxy-corn starches with varying degrees of substitution by carboxyl and acetyl groups in our research. The process of enzymatic hydrolysis was performed in a recirculated membrane reactor (CRMR). Our findings indicated that the physicochemical properties of starch derivatives and their hydrolysates depended on the biological origin of the biopolymer and the type and degree of modification. However, the presence of carboxyl groups in the modified starch molecules is critical and affects the rheological properties and water-binding capacity of the starch preparations. For example, in the case of waxy-corn starch preparations with a lower content of carboxyl groups (i.e., derivatives with a low degree of oxidation), the water-binding capacity (WBC) increases when compared to native starch. The highest WBC value of 206.3% was noted for the doubly modified waxy-corn starch with an oxidation degree of 0.2% and an acetylation degree of 2.5%, while native waxy-corn starch shows a WBC of 161.4%. In contrast, it was observed that preparations with a higher content of carboxyl groups, i.e., derivatives with an oxidation degree of 2.5%, show a lower swelling power compared to native waxy starch.

Comparisons of the amylolytic enzymes and malt starch hydrolysates of two barley cultivars, Hokudai 1 (the first cultivar developed in Japan) and Kitanohoshi (currently used cultivar for beer production).

Starch degradation in malted barley produces yeast-fermentable sugars. In this study, we compared the amylolytic enzymes and composition of the malt starch hydrolysates of two barley cultivars, Hokudai 1 (the first cultivar established in Japan) and Kitanohoshi (the currently used cultivar for beer production). Hokudai 1 malt contained lower activity of amylolytic enzymes than Kitanohoshi malt, although these cultivars contained α-amylase AMY2 and β-amylase Bmy1 as the predominant enzymes. Malt starch hydrolysates of Hokudai 1 contained more limit dextrin and less yeast-fermentable sugars than that of Kitanohoshi. In mixed malt saccharification, a high Hokudai 1 malt ratio increased the limit dextrin levels and decreased the maltotriose and maltose levels. Even though Kitanohoshi malt contained more amylolytic enzymes than Hokudai 1 malt, addition of Kitanohoshi extract containing the amylolytic enzymes did not enhance malt starch degradation of Hokudai 1. Hokudai 1 malt starch was less degradable than Kitanohoshi malt starch.

Isolated cassava cells: Comparison of structure and physicochemical properties with starch and whole flour

Individual cells are the smallest units of the plant tissue structure, and their structure and physicochemical properties are essential for whole food processing. In this study, cassava cells were isolated using acid-alkali, hydrothermal, and pectinase methods, and the differences in microstructure and physicochemical properties among the cells, starch, and whole flour were investigated. Cassava cells isolated using pectinase showed intact individual cells with a higher isolation rate and less damage to the cell wall structure and intracellular composition. The presence of cell walls in intact individual cells inhibited the swelling and leaching of starch, resulting in a lower peak viscosity and higher gelatinization temperature than those of starch. The intact cell structure and non-starch composition enhanced the shear resistance of the gels in the sample. Heat treatment led to the gelatinization of intracellular starch and increased the permeability of the cell wall, destroying the physical barrier function of the cell wall; however, the compact cell matrix and non-starch components can inhibit starch hydrolysis. This study suggests that cells isolated using pectinase can be used to investigate the effect of cell walls on the functional properties of intracellular starch in cassava. The isolated cells provide new insights into the cassava industry.

Effects of adding blueberry residue powder and extrusion processing on microstructure and in vitro digestibility of indica rice flour

Using crushed indica rice flour as the main raw material, after adding blueberry residue powder, the indica rice puffed powder containing blueberry residue (IREPBR) was obtained through extrusion and crushing. The effects of adding blueberry residue and extrusion processing on the microstructure and in vitro digestibility of indica rice flour were studied. The basic component analysis results showed that the fat and total starch content of indica rice powder significantly decreased after adding blueberry residue powder and extrusion puffing, while the total dietary fiber content significantly increased. The results of scanning electron microscope (SEM) showed that there were many pits and bubbles in the extruded indica rice flour after adding blueberry residue powder. Compared with indica rice flour (IRF), its Water Absorption Index (WAI) and Water Solubility Index (WSI) significantly increased, and its thermal properties also changed. XRD and FT-IR analysis showed that after extrusion, the crystalline morphology of indica rice starch changed from A-type to V-type, and the relative crystallinity also changed. In addition, the content of resistant starch (RS) in indica rice flour added with blueberry residue powder and processed by extrusion increased significantly, while the starch hydrolysis rate and estimated blood glucose index (eGI) decreased.

Engineering Bacillus licheniformis as industrial chassis for efficient bioproduction from starch

Starch is an attractive feedstock in biorefinery processes, while the low natural conversion rate of most microorganisms limits its applications. Herein, starch metabolic pathway was systematically investigated using Bacillus licheniformis DW2 as the host organism. Initially, the effects of overexpressing amylolytic enzymes on starch hydrolysis were evaluated. Subsequently, the transmembrane transport system and intracellular degradation module were modified to accelerate the uptake of hydrolysates and their further conversion to glucose-6-phosphate. The DW2-derived strains exhibited robust growth in starch medium, and productivity of bacitracin and subtilisin were improved by 38.5% and 32.6%, with an 32.3% and 22.9% increase of starch conversion rate, respectively. Lastly, the employment of engineering strategies enabled another B. licheniformis WX-02 to produce poly-γ-glutamic acid from starch with a 2.1-fold increase of starch conversion rate. This study not only provided excellent B. licheniformis chassis for sustainable bioproduction from starch, but shed light on researches of substrate utilization.

Cell wall permeability in relation to in vitro starch digestion of pea cotyledon cells

The role of cell wall permeability and rate of starch digestibility in intact cotyledon cells from four different varieties of pea seeds was studied. Pulsed-field gradient nuclear magnetic resonance (PFG NMR) coupled with light and confocal microscopy were employed to evaluate the cotyledon cells' diffusion coefficients and cell wall permeability. The cotyledon cells' diffusion coefficients and cell wall permeability followed a decreasing trend: White/yellow pea > Marrowfat pea > Maple pea > Blue pea. The varying size of internal cavities in the microstructure in the cotyledon cells, as observed by the light and confocal micrographs, may be responsible for this trend. The extent of starch hydrolysis recorded from the cotyledon cells followed the same trend of the cell wall permeability except for Blue pea cotyledon cells. Thus, indicating that the more permeable the cotyledon cell to the starch-degrading enzymes, the higher the extent of intracellular starch hydrolysis. The microstructure changes in the cotyledon cells during digestion also confirmed this observation.

Safety evaluation of the food enzyme lysophospholipase from the genetically modified Trichoderma reesei strain DP-Nyc81.

The food enzyme lysophospholipase (2-lysophosphatidylcholine acylhydrolase, EC 3.1.1.5) is produced with the genetically modified Trichoderma reesei strain DP-Nyc81 by Genencor International B.V. The genetic modifications do not give rise to safety concerns. The food enzyme is free from viable cells of the production organism and its DNA. It is intended to be used in the processing of cereals and other grains for the production of glucose syrups and other starch hydrolysates. Since residual amounts of food enzyme-total organic solids are removed during these food manufacturing processes, dietary exposure was not calculated and toxicological studies were considered unnecessary. A search for the identity of the amino acid sequence of the food enzyme to known allergens was made and no match was found. The Panel considered that the risk of allergic reactions upon dietary exposure cannot be excluded, but the likelihood is low. Based on the data provided, the Panel concluded that this food enzyme does not give rise to safety concerns, under the intended conditions of use.

Artificial antibody-antigen-directed immobilization of α-amylase to hydrolyze starch for cascade reduction of 2-nitro-4-methylphenol to 2-amino-4-methylphenol

Nitrophenol is a hazardous substance that poses a threat to the environment and human health, and its treatment has attracted widespread attention. The purpose of this study is to establish an environmentally friendly α-amylase system for the hydrolysis of starch to reduce nitrophenol to aminophenol through cascade reactions. The α-amylase system was obtained through artificial antibody-antigen-directed immobilization, including the synthesis of artificial antibodies, synthesis of artificial antigens, and affinity assembly. In this process, catechol and protocatechuic aldehyde were used to prepare artificial antibodies and artificial antigens respectively through polymerization and Schiff base reactions. Then, artificial antibodies captured the catechol in the artificial antigen structure to form immobilized α-amylases. Compared with free α-amylase, the immobilized α-amylase showed a good reusability and excellent regenerative ability. Subsequently, the immobilized α-amylase were used in the reaction of catalyzing starch hydrolysis to synthesize 2-amino-4-methylphenol, and the yield of 2-amino-4-methylphenol was 58.88 ± 0.19 %. After 5 consecutive catalytic reactions, a yield of 47.61 ± 1.27 % can still be achieved.

Production of Amylase Enzyme by Aspergillus and Fusarium Species using Sugar Cane Bagasse

Study’s Novelty/Excerpt This study presents insights into the cost-effective production of amylase enzymes using sugar cane bagasse as a substrate, showcasing a significant advancement over traditional synthetic starch substrates. By optimizing production parameters such as incubation time, substrate and inoculum concentrations, pH, and temperature, the research demonstrates the superior amylase yield of Aspergillus niger, Aspergillus flavus, and Fusarium spp. under specific conditions. This work not only highlights the potential of using waste materials for industrial enzyme production but also emphasizes the need for further exploration of the isolates' identities and the industrial applications of the extracted enzymes. Full Abstract Amylases are hydrolytic enzymes produced by some microorganisms and utilized in the hydrolysis of starch. This study aimed to determine the ability of certain fungal isolates isolated from wastes to synthesize the amylase enzyme utilizing synthetic soluble starch and sugar cane bagasse as substrates. Aspergillus niger, Aspergillus flavus, and Fusarium spp., previously identified to have amylolytic activity, were used for the research. The submerged fermentation process was used to produce amylase, with basal medium and sugar cane bagasse as substrates. The incubation time, substrate and inoculum concentrations, pH, and temperature were all optimized. The dinitrosalicylic acid reagent (DNS) technique was used to determine the activity of the amylase generated. Initial production of amylase using soluble starch (20 g (w/v)) as substrate at room temperature and pH 7.0 shows that all the isolates produce amylase better when their concentration (3%) is high but have different incubation periods of 96 hr for Aspergillus niger (8.65±0.21 U/mL/minutes) and Fusarium spp (7.15±0.07 U/mL/minutes), while Aspergillus flavus (7.30±0.14 U/mL/minutes) requires an extended incubation period of 144 hr to produce the product. Further production using sugar cane bagasse and optimization of production parameters of the isolates reveals that Aspergillus niger (4.35±0.07 U/mL/minutes) has an optimum incubation period of 120 hours, an inoculum concentration and substrate concentration of 2% each, and a pH of 6, Aspergillus flavus (6.40±0.28 U/mL/minutes) has an optimum incubation period of 144 hours, with inoculum and substrate concentration of 3% each at neutral pH, Fusarium spp (6.80±0.28 U/mL/minutes) has an optimum incubation period of 168hr., inoculum concentration of 3%, substrate concentration of 2% and a neutral pH-7 and all the isolates have their best yield at temperature of 30oC. In conclusion, using cheap and readily available sugar cane bagasse can be a more cost-effective option for expensive synthetic starch substrates used in amylase synthesis. The study suggests further investigation into the isolates' identities and the extracted enzyme's industrial application.