Modulation of fried spring roll wrapper quality upon treatment of batter with maltogenic amylase, transglutaminase and bromelain.

BACKGROUND: Fried foods are favored for their unique crispiness, golden color and flavor, but they also face great challenge because of their high oil content, high calories and the existence of compounds such as acrylamide and polycyclic aromatic hydrocarbons. Long-term consumption of fried foods may adversely affect health. Therefore, it is necessary to explore fried foods with lower oil contents and a high quality to meet the demand. RESULTS: A method of enzyme treatment was explored to investigate the effects of maltogenic amylase (MA), transglutaminase (TG) and bromelain (BRO) on the physicochemical properties of the batter and the quality of fried spring roll wrapper (FSRW). The results showed that the MA-, TG- or BRO-treated batters had a significant shear-thinning behavior, especially with an increase in viscosity upon increasing TG contents. FSRW enhanced its fracturability from 419.19 g (Control) to 616.50 g (MA-6 U g-1), 623.49 g (TG-0.75 U g-1) and 644.96 g (BRO-10 U g-1). Meanwhile, in comparison with BRO and MA, TG-0.5 U g-1 endowed batter with the highest density and thermal stability. MA-15 U g-1 and TG-0.5 U g-1 displayed FSRW with uniform and dense pores, and significantly reduced its oil content by 18.05% and 25.02%, respectively. Moreover, compared to MA and TG, BRO-50 U g-1 improved the flavor of FSRW. CONCLUSION: MA, TG or BRO played a key role in affecting the physicochemical properties of the batter and the quality of FSRW. TG-0.5 U g-1 remarkly reduced the oil content of FSRW with a great potential in practical application. © 2024 Society of Chemical Industry.

Function of the mdxR gene encoding a novel regulator for carbohydrate metabolism and sporulation in Bacillus subtilis 168.

The mdxR gene located upstream of mdxD, encoding a maltogenic amylase, has been annotated as a member of LacI-type transcriptional regulator in Bacillus subtilis 168 but its function has not been investigated yet. In this study, expression pattern of the mdxR promoter (PmdxR) and effects of mdxR were investigated to elucidate the function of mdxR. Expression of PmdxR was monitored by the ß-galactosidase activity expressed from the PmdxR-lacZ fusion integrated at the amyE locus on the chromosome. The promoter was induced by starch, ß-cyclomaltodextrin, or maltose at early exponential phase and kept expressed until late stationary phase. However, it was repressed by glucose, sucrose, or glycerol, suggesting that it was under catabolite repression. Furthermore, interactions of MdxR and Spo0A to the DNA fragment carrying PmdxR or PmdxD were detected by mobility-shift assay, implying that MdxR was a novel transcription regulator for both genes, which were regulated also by Spo0A. The mdxR mutant impaired the expressions of mdxD and malL (encoding an α-glucosidase); degraded accumulated glycogen slower than the wild type and the mdxD mutant. Both of the mdxR and the mdxD mutants formed more endospores (50.95% and 47.10%) than the wild type (23.90%). Enhanced sporulation by these mutations could be of industrial interest where sporulation or endospores of B. subtilis matters. These results indicate that MdxR functions as a transcriptional regulator for mdxR, mdxD, and other genes in the gene cluster that is related to the maltose/maltodextrin metabolism. MdxR and MdxD are also involved in glycogen metabolism and sporulation, tentatively by modulating the net energy balance in the cell.

Conversion of waste cooked rice into maltose by a Bacillus subtilis-derived maltogenic amylase and production of maltobionic acid from the prepared maltose using genetically modified Pseudomonas taetrolens.

In this study, we attempted to produce maltobionic acid (MBA) from waste cooked rice (WCR) using maltose as an intermediate. In our previous study, we produced maltose from WCR using a commercial maltogenic amylase (Maltogenase L). However, in the present study, we used wild-type Bacillus subtilis, which inherently produces maltogenic amylase (AmyE), instead of Maltogenase L to produce maltose from WCR. During cultivation of B. subtilis with WCR, maltose was successfully produced by AmyE in the culture medium. To improve maltose production, we constructed a recombinant B. subtilis strain expressing AmyE and used it for maltose production. Following cultivation of the recombinant B. subtilis strain, the maltose production titer (34.6 g/L) increased approximately 3.6-fold that (9.6 g/L) obtained from the cultivation of wild-type B. subtilis. Using Pseudomonas taetrolens, an efficient MBA-producing bacterium, 28.8 g/L of MBA was produced from the prepared maltose (27.6 g/L). The above results indicated that MBA was successfully produced from WCR via a two-step process, which involved the conversion of WCR into maltose by maltogenic amylase-producing B. subtilis and the production of MBA from the WCR-derived maltose by P. taetrolens.

Phenylalanine 314 is essential for the activity of maltogenic amylase from Corallococcus sp. EGB.

Maltogenic amylase CoMA from Corallococcus sp. strain EGB catalyzes the hydrolysis and transglycosylation of maltooligosaccharides and soluble starch into maltose, the sole hydrolysate. This process yields pure maltose with potentially wide applications. Here, we identified and evaluated the role of phenylalanine 314 (F314), a key amino acid located near the active center, in the catalytic activities of the CoMA. Site-directed mutagenesis analysis showed that the activity of a F314L mutant on potato starch substrate decreased to 26% of that of wild-type protein. Compared with the wild-type, F314L exhibited similar substrate specificity, hydrolysis pattern, pH, and temperature requirements. Circular dichroism spectrum data showed that the F314L mutation did not affect the structure of the folded protein. In addition, kinetic analysis demonstrated that F314L exhibited an increased Km value with lower substrate affinity. Homology modeling showed that the benzene ring structure of F314L was involved in π-π conjugation, which might potentially affect the affinity of CoMA toward starch. Taken together, these data demonstrated that F314 is essential for the hydrolytic activity of the CoMA from Corallococcus sp. strain EGB.

High level food-grade expression of maltogenic amylase in Bacillus subtilis through dal gene auxotrophic selection marker.

As a food-safe microorganism, Bacillus subtilis has been widely utilized in the production of food enzyme, where a food-grade expression system without antibiotic is required. However, there is no mature system for such expression, since the recombinant plasmid in existing food-grade expression system is unstable especially in high-density fermentation. In this study, we constructed a food-grade expression system based on the dal gene auxotrophic selection marker. Specifically, maltogenic amylase (AmyM) was expressed in dal deletion strain without antibiotic, yielding an activity of 519 U/mL. To increase the expression of AmyM, the promoter of amyM (gene encoding AmyM) was optimized. Furthermore, we found that excessive expression of dal gene was detrimental to the stability of plasmid, and the ribosome binding site (RBS) of dal was mutated with the reduced synthesis of D-alanine. After that, AmyM activity increased to 1364 U/mL with the 100 % stability of plasmid. The 3-L fermentor cultivation was performed with the highest value ever reported in food-grade microorganisms, an activity of 2388 U/mL, showing the scale-up production capability of this system. Besides, it is also able to apply the system for other food enzymes, which indicating the great generalizability of this system for different application.

Safety evaluation of the food enzyme glucan 1,4-α-maltohydrolase from the genetically modified Bacillus subtilis strain BABSC.

The food enzyme glucan 1,4-α-maltohydrolase (4-α-d-glucan α-maltohydrolase, EC 3.2.1.133) is produced with the genetically modified Bacillus subtilis strain BABSC by Advanced Enzyme Technologies Ltd. The requirements for the qualified presumption of safety (QPS) approach have not been met. The food enzyme is free from viable cells of the production organism and its DNA. It is intended to be used in baking processes and starch processing for the production of glucose syrups and other starch hydrolysates. Since residual amounts of total organic solids (TOS) are removed, dietary exposure was not calculated for starch processing for the production of glucose syrups and other starch hydrolysates. For baking processes, the dietary exposure was estimated to be up to 0.101 mg TOS/kg body weight per day in European populations. No toxicological studies were provided by the applicant. A search for the similarity of the amino acid sequence of the food enzyme to known allergens was made and one match with a respiratory allergen was found. The Panel considered that the risk of allergic reactions by dietary exposure cannot be excluded, but the likelihood is low. In the absence of appropriate data to fully characterise the production strain, the Panel was unable to conclude on the safety of the food enzyme under the intended conditions of use.

Effect of modification by maltogenic amylase and branching enzyme on the structural and physicochemical properties of sweet potato starch.

Sweet potato starch (SPSt) was treated sequentially with the combination of maltogenic amylase (MA) and branching enzyme (BE) (MA â†’ BE) or BE and MA (BE→MA) to modify its structural and physicochemical properties. Following the MA â†’ BE and BE→MA modifications, the degree of branching was increased from 12.02 % to 44.06 %; whereas, the average chain length (ACL) decreased from 18.02 to 12.32. Fourier-transform infrared spectroscopy and digestive performance analysis indicated that the modifications reduced hydrogen bonds and increased resistant starch in SPSt. Rheological analysis revealed that the storage and loss moduli of the modified samples were lower than those of the control samples, except for starch treated with MA alone. X-ray diffraction measurements suggested that the re-crystallisation peak intensities of the enzyme-modified starches were lower than those of the untreated sample. The retrogradation resistance ability of the analysed samples followed the order: BE→MA-starches > MA â†’ BE-starches > untreated starch. The relationship between the crystallisation rate constant and short branched chains (DP6-9) was well described by linear regression. This study provides a theoretical foundation for retarding the retrogradation of starch, which can improve food quality and extend the shelf-life of enzymatically modified starchy foods.

Maltogenic amylase: Its structure, molecular modification, and effects on starch and starch-based products.

Maltogenic amylase (MAA) (EC3.2.1.133), a member of the glycoside hydrolase family 13 that mainly produces α-maltose, is widely used to extend the shelf life of bread as it softens bread, improves its elasticity, and preserves its flavor without affecting dough processing. Moreover, MAA is used as an improver in flour products. Despite its antiaging properties, the hydrolytic capacity and thermal stability of MAA can't meet the requirements of industrial application. However, genetic engineering techniques used for the molecular modification of MAA can alter its functional properties to meet application-specific requirements. This review briefly introduces the structure and functions of MAA, its application in starch modification, its effects on starch-based products, and its molecular modification to provide better insights for the application of genetically modified MAA in starch modification.

Characterization of an Amylolytic Enzyme from Massilia timonae of the GH13_19 Subfamily with Mixed Maltogenic and CGTase Activity.

This work reports the characterization of an amylolytic enzyme from the bacteria Massilia timonae CTI-57. A gene encoding this protein was expressed from the pTrcHis2B plasmid in Escherichia coli BL21 Star™ (DE3). The purified protein had 64 kDa, and its modeled structure showed a monomer with the conserved α-amylases structure composed of the domain A with the characteristic (ß/α)8-barrel, the small domain B, and the domain C with an antiparallel beta-sheet. Phylogenetic analysis demonstrated that the expressed protein belongs to the GH13_19 subfamily of glycoside hydrolases. The ions Ca2+, Mn2+, Na+, Mg2+, Mo6+, and K+ did activate the purified enzyme, while EDTA and the ions Fe2+, Hg2+, Zn2+, and Cu2+ were strong inhibitors. SDS was also a strong inhibitor. The enzyme's optimal pH and temperature were 7.0 and 45 °C, respectively, and its Tm was 62.2 °C. The KM of the purified enzyme for starch was 13 mg/mL, and the Vmax was 0.24 µmol of reducing sugars released per min. The characterized enzyme presented higher specificity for maltodextrin and starch and produced maltose as the main starch hydrolysis product. This is the first characterized maltose-forming amylolytic enzyme from the GH13_19 subfamily. The purified enzyme produced ß-cyclodextrin from starch and maltodextrin and could be considered a cyclodextrin glucanotransferase (CGTase). This is the first report of a GH13_19 subfamily enzyme with CGTase activity.

Safety evaluation of the food enzyme glucan 1,4-α-maltohydrolase from the genetically modified Bacillus subtilis strain AR-513.

The food enzyme glucan 1,4-α-maltohydrolase (4-α-D-glucan α-maltohydrolase EC 3.2.1.133) is produced with the genetically modified Bacillus subtilis strain AR-513 by AB Enzymes GmbH. 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 four food manufacturing processes: processing of cereals and other grains for the production of baked products, other cereal-based products and glucose syrups and other starch hydrolysates; processing of plant- and fungal-derived products for the production of plant-based analogues of milk and milk products. Since residual amounts of total organic solids (TOS) are removed in the production of glucose syrups and other starch hydrolysates, dietary exposure was not calculated for this process. For the remaining three food processes, exposure to the food enzyme-TOS was estimated to be up to 0.2 µg TOS/kg body weight per day in European populations. As the production strain of B. subtilis strain AR-513 meets the requirements for the qualified presumption of safety (QPS) approach and no issue of concern arose from the production process, no toxicological data were required. A search for the similarity of the amino acid sequence of the food enzyme to known allergens was made and five matches with respiratory allergens and one with a mosquito allergen were found. The Panel considered that, under the intended conditions of use, the risk of allergic reactions by dietary exposure cannot be excluded, but the likelihood is low. Based on the data provided, the Panel concluded that this food enzyme did not give rise to safety concerns, under the intended conditions of use.

Safety evaluation of the food enzyme glucan 1,4-α-maltohydrolase from the genetically modified Bacillus licheniformis strain NZYM-SD.

The food enzyme glucan 1,4-α-maltohydrolase (4-α-d-glucan α-maltohydrolase; 3.2.1.133) is produced with the genetically modified Bacillus licheniformis strain NZYM-SD by Novozymes A/S. The genetic modifications did not give rise to safety concerns. The production strain has been shown to qualify for Qualified Presumption of Safety (QPS) status. The food enzyme is free from viable cells of the production organism and its DNA. The food enzyme is intended to be used in three food manufacturing processes, namely baking processes and brewing processes and starch processing for glucose syrup production and other starch hydrolysates. Since residual amounts of total organic solids (TOS) are removed by the purification steps applied during the production of glucose syrups, dietary exposure was calculated only for baking and brewing processes. Dietary exposure was estimated to be up to 0.57 mg TOS/kg body weight (bw) per day in European populations. Given the QPS status of the production strain and the lack of hazards resulting from the food enzyme manufacturing process, toxicological studies were not considered necessary. Similarity of the amino acid sequence to those of known allergens was searched and four matches were found. The Panel considered that, under the intended conditions of use, the risk of allergic sensitisation and elicitation reactions upon dietary exposure to this food enzyme cannot be excluded, but the likelihood of such reactions to occur is considered to be low. Based on the data provided, the QPS status of the production strain and the absence of issues arising from the production process, the Panel concluded that the food enzyme glucan 1,4-α-maltohydrolase produced with the genetically modified B. licheniformis strain NZYM-SD does not give rise to safety concerns under the intended conditions of use.

Safety evaluation of the food enzyme glucan 1,4-α-maltohydrolase from the genetically modified Bacillus licheniformis strain NZYM-CY.

The food enzyme glucan 1,4-α-maltohydrolase (4-α-d-glucan α-maltohydrolase; 3.2.1.133) is produced with the genetically modified Bacillus licheniformis strain NZYM-CY by Novozymes A/S. The genetic modifications did not give rise to safety concerns. The production strain has been shown to qualify for Qualified Presumption of Safety (QPS) status. The food enzyme is free from viable cells of the production organism and its DNA. The food enzyme is intended to be used in three food manufacturing processes, namely baking and brewing processes and starch processing for glucose syrup production and other starch hydrolysates. Since residual amounts of total organic solids (TOS) are removed by the purification steps applied during the production of glucose syrups, dietary exposure was calculated only for the baking and brewing processes. Dietary exposure was estimated to be up to 0.45 mg TOS/kg body weight (bw) per day in European populations. Given the QPS status of the production strain and the lack of hazards resulting from the food enzyme manufacturing process, toxicological studies were not considered necessary. Similarity of the amino acid sequence to those of known allergens was searched and four matches were found. The Panel considered that, under the intended conditions of use, the risk of allergic sensitisation and elicitation reactions upon dietary exposure to this food enzyme cannot be excluded, but the likelihood of such reactions to occur is considered to be low. Based on the data provided, the QPS status of the production strain and the absence of issues arising from the production process, the Panel concluded that the food enzyme glucan 1,4-α-maltohydrolase produced with the genetically modified B. licheniformis strain NZYM-CY does not give rise to safety concerns under the intended conditions of use.

Safety evaluation of the food enzyme glucan 1,4-α-maltohydrolase from the genetically modified Bacillus licheniformis strain NZYM-FR.

The food enzyme glucan 1,4-α-maltohydrolase (4-α-d-glucan α-maltohydrolase; 3.2.1.133) is produced with the genetically modified Bacillus licheniformis strain NZYM-FR by Novozymes A/S. The genetic modifications did not give rise to safety concerns. The production strain has been shown to qualify for Qualified Presumption of Safety (QPS) status. The food enzyme is free from viable cells of the production organism and its DNA. The food enzyme is intended to be used in three food manufacturing processes, namely baking and brewing processes and starch processing for glucose syrup production and other starch hydrolysates. Since residual amounts of total organic solids (TOS) are removed by the purification steps applied during the production of glucose syrups, dietary exposure was calculated only for the baking and brewing processes. Dietary exposure was estimated to be up to 0.30 mg TOS/kg body weight (bw) per day in European populations. Given the QPS status of the production strain and the lack of hazards resulting from the food enzyme manufacturing process, toxicological studies were not considered necessary. Similarity of the amino acid sequence to those of known allergens was searched and four matches were found. The Panel considered that, under the intended conditions of use, the risk of allergic sensitisation and elicitation reactions upon dietary exposure to this food enzyme cannot be excluded, but the likelihood of such reactions to occur is considered to be low. Based on the data provided, the QPS status of the production strain and the absence of issues arising from the production process, the Panel concluded that the food enzyme glucan 1,4-α-maltohydrolase produced with the genetically modified B. licheniformis strain NZYM-FR does not give rise to safety concerns under the intended conditions of use.

Improving Aqueous Solubility of Natural Antioxidant Mangiferin through Glycosylation by Maltogenic Amylase from Parageobacillus galactosidasius DSM 18751.

Mangiferin is a natural antioxidant C-glucosidic xanthone originally isolated from the Mangifera indica (mango) plant. Mangiferin exhibits a wide range of pharmaceutical activities. However, mangiferin's poor solubility limits its applications. To resolve this limitation of mangiferin, enzymatic glycosylation of mangiferin to produce more soluble mangiferin glucosides was evaluated. Herein, the recombinant maltogenic amylase (MA; E.C. 3.2.1.133) from a thermophile Parageobacillus galactosidasius DSM 18751T (PgMA) was cloned into Escherichia coli BL21 (DE3) via the expression plasmid pET-Duet-1. The recombinant PgMA was purified via Ni2+ affinity chromatography. To evaluate its transglycosylation activity, 17 molecules, including mangiferin (as sugar acceptors), belonging to triterpenoids, saponins, flavonoids, and polyphenol glycosides, were assayed with ß-CD (as the sugar donor). The results showed that puerarin and mangiferin are suitable sugar acceptors in the transglycosylation reaction. The glycosylation products from mangiferin by PgMA were isolated using preparative high-performance liquid chromatography. Their chemical structures were glucosyl-α-(1→6)-mangiferin and maltosyl-α-(1→6)-mangiferin, determined by mass and nucleic magnetic resonance spectral analysis. The newly identified maltosyl-α-(1→6)-mangiferin showed 5500-fold higher aqueous solubility than that of mangiferin, and both mangiferin glucosides exhibited similar 1,1-diphenyl-2-picrylhydrazyl free radical scavenging activities compared to mangiferin. PgMA is the first MA with glycosylation activity toward mangiferin, meaning mangiferin glucosides have potential future applications.

An enzyme-mediated controlled release system for curcumin based on cyclodextrin/cyclodextrin degrading enzyme.

In this study, an enzyme-triggered system based on ß-cyclodextrin (ß-CD) has been developed to achieve controlled release of hydrophobic drugs in the presence of maltogenic amylase (MAase). The inclusion complex formation of curcumin (CUR), as a model anticancer compound, with ß-CD was characterized by fluorescence and Fourier transform infrared (FTIR) spectroscopy. CUR was loaded into ß-CD with an encapsulation efficiency of approximately 30 %. The in vitro profiles of CUR release from ß-CD showed that 100 % of the drug was released after one hour incubation in the presence of MAase with cyclodextrin degrading activity. Fluorescence microscopy images indicate a significantly greater cellular uptake of CUR using ß-CD-CUR/MAase system compared to ß-CD-CUR inclusion complex without MAase. The ß-CD-CUR/MAase system exhibited lower IC50 values and greater anti-proliferative effects in comparison with free CUR and ß-CD-CUR in MCF-7 and Huh-7 cancer cells. The results from fluorescence microscopy and flow cytometric assay using the acridine orange/ethidium bromide and Annexin V-PE/7-AAD staining suggest that the ß-CD-CUR/MAase system exhibited higher cytotoxic and apoptotic effects on cancer cells compared to other formulations. This triggered release of CUR in the presence of MAase is owing to the ß-CD degradation by MAase resulting ring opening and chain scission in ß-CD. We demonstrate that this enzyme-mediated controlled release system has a potential application for controlled release of poorly water-soluble drugs or hydrophobic compounds such as CUR.

A new maltogenic amylase from Bacillus licheniformis R-53 significantly improves bread quality and extends shelf life.

Maltogenic amylase suppressed starch retrogradation in baked products. Here, a maltogenic amylase-producing strain of bacteria was screened and identified as Bacillus licheniformis R-53. Its coding gene was cloned and over-expressed in Bacillus subtilis WB600. Recombinant maltogenic amylase BLMA exhibited activity of 3235 U/mg under optimal conditions (60 °C and pH 6.5), with a good thermostability and pH stability. Mixolab experiment showed that a concentration of 60 ppm BLMA significantly improved the operating characteristics of dough. Baking test indicated the recombinant BLMA reduced bread hardness by 2.12 times compared with the control. Compared with maltogenic amylase from Novozymes (Novamyl 3D BG) and Angel Yeast Co. Ltd. (MAM100), BLMA has better effect on improving the bread volume, and almost the same effect on reducing hardness, improving elasticity and maintaining sensory as Novamyl 3D BG. Adding BLMA improved bread quality, increased bread volume and decreased hardness during storage, thus extending its shelf life.

Influence of germinated brown rice-based flour modified by MAse on type 2 diabetic mice and HepG2 cell cytotoxic capacity.

This study aimed to discover whether using maltogenic amylase (MAse) to modify starch in germinated brown rice flour may enhance slow digestion starch and release more bioactive compounds (BCs) content. To achieve this aim, the starch was modified with four levels of MAse (0 U, 133 U, 266 U and 399 U MAse/g flour) for 1 hr at pH 5 and then spray-dried to make modified flour. The biochemical impacts of the products were then accessed in normal and type 2 diabetic mice for 4 weeks. The result showed that when the starch was modified by MAse 266 U/g, a significant reduction of rapidly digested starch to 22.35% from 61.56%, an increase in slowly digested starch to 33.09% while resistant starch as 2.92% corresponding to the increase of γ-amino butyric acid to 528.1 ± 44.1 mg/L and 120.6 ± 10.9 mg/L of ferulic acid. The extract from modified flour showed very strong cytotoxic activity against HepG2 cell (>80% inhibition). The result in vivo showed that the type-2 diabetic mice fed with this modified product could better improve the stability of the glycemic index. Also, atherosclerotic plaque assessment further supports these findings. The results indicated that BCs released considerably couple with the changes in starch properties caused by MAse enhanced the effectiveness of this product to diabetes as well as positive effect on cytotoxic activity against HepG2 cell.

Investigation of starch functionality and digestibility in white wheat bread produced from a recipe containing added maltogenic amylase or amylomaltase.

In the crumb of fresh white wheat bread, starch is fully gelatinized. Its molecular and three-dimensional structure are major factors limiting the rate of its digestion. The aim of this study was to in situ modify starch during bread making with starch-modifying enzymes (maltogenic amylase and amylomaltase) and to investigate the impact thereof on bread characteristics, starch retrogradation and digestibility. Maltogenic amylase treatment increased the relative content of short amylopectin chains (degree of polymerization ≤ 8). This resulted in lower starch retrogradation and crumb firmness upon storage, and reduced extent (up to 18%) of in vitro starch digestion for fresh and stored breads. Amylomaltase only modestly shortened amylose chains and had no measurable impact on amylopectin structure. Modification with this enzyme led to slower bread crumb firming but did not influence starch digestibility.

Optimization of encapsulation of maltogenic amylase into a mixture of maltodextrin and beeswax and its application in gluten-free bread.

In this work maltogenic amylase (MAase) was encapsulated into the mixture of maltodexrin and beeswax (BW) for retarding staling of gluten-free bread. The effects of maltogenic amylase (MAase) concentration (8.2, 45, and 82 mg/ml), maltodextrin with dextrose equivalent (DE) 4-7 (MD) (1, 2.5, and 4%) and, BW (1, 2.5, and 4%) on the encapsulation efficiency (EE) of encapsulated enzyme were optimized using RSM. The optimized formulation was MAase with 8.2 mg/ml, MD 4% and BW 1%, leading to the highest EE (79.35%), thus chosen for subsequent experiments. The prepared particles were 1,190.50 nm with PDI of 0.336 and zeta potential of -8.30 mV. Surface morphologies of produced particles were almost spherical with layered appearance. Batter with this formulation led to higher cross over point in frequency sweep than free enzyme-loaded batter. Lower weight loss, higher volume index, darker crust color, whiter crumb color, more aerated microstructure, less hardness in crumb, and higher sensorial acceptability on the first day and during storage period in the breads containing encapsulated MAase was observed.

Impact of maltogenic α-amylase on the structure of potato starch and its retrogradation properties.

Structural modification of starch using efficient α-amylases to improve its properties is an established method in the starch industry. In our previous research, the novel maltogenic α-amylase CoMA that catalyzes multi-molecular reactions has been identified. In this study, the impact of CoMA on the structure and retrogradation properties of potato starch was evaluated. CoMA cleaves internal starch chains to change the proportion of amylose and amylopectin in starch. Following treatment, visible pores and microporous on the surface of starch granules were observed from SEM analysis. CoMA modification led to increased insoluble blue complex formation and hydrolysis to shorten the outer chains, which was found to reduce the development rate of starch according to network interactions from the dynamic rheological analysis. Furthermore, maltose accumulation with water competition was also deduced to be involved in the inhibition of retrogradation. Its activities in the cleavage of internal starch granules, shortening of outer chains of starch, and maltose formation make CoMA a powerful agent for the inhibition of starch retrogradation with a very low effective dose of 0.5 mg/kg, which may find potential applications in the starch processing industry.