Physicochemical,crystalline, and thermal properties of native,oxidized, acid,and enzyme hydrolyzed Chinese yam (Dioscorea opposite Thunb) starch

Native yam starch was modified by oxidation (with sodium hypochlorite), acid, and enzyme (with gluco‐amylase), respectively. Ash content, fat content, protein, and AM contents were reduced following modifications. All the modifications reduced swelling capacity whereas improved the solubility. The SEM of the native starch showed the presence of large oval or spherical to small irregular‐shaped granules, however, oxidation and hydrolysis caused collapse of the granules. From the XRD, the typical C‐type XRD pattern of the native starch changed to the typical A‐type XRD pattern of the three modified starches. The above results also demonstrated that amorphous region of yam starch granules was preferentially degraded than crystalline region of granules. DSC showed that oxidation and acid/enzyme hydrolysis reduced peak temperature of gelatinization (T_p) of native starch significantly. Enthalpy of gelatinization (ΔH) of oxidation and enzyme starch was reduced but it has no obviously change following acid modification.     

Visual Observation of Hydrolyzed Potato Starch Granules by α‐Amylase with Confocal Laser Scanning Microscopy

Porous starch was produced by digestion of freeze‐dried potato starch with α‐amylase from Bacillus sp. The surface structure of the granules became perforated and in the interior of the granules a capsule‐like cavity was formed, i.e. the hydrolyzed starch can be used as an encapsulant. The structure change of the granules was observed with confocal laser scanning microscopy and scanning electron microscopy. The degree of starch hydrolysis could be correlated with the Avrami equation. The activation energy of starch hydrolysis by α‐amylase was 83 kJ/mol.     

Physicochemical,crystalline, and thermal properties of native and enzyme hydrolyzed Pueraria lobata (Willd.) Ohwi and Pueraria thomsonii Benth. starches

Starches isolated from the bulbs of Pueraria lobata (Willd.) Ohwi (PLO) and Pueraria thomsonii Benth. (PTB) were hydrolysed by glucoamylase for different lengths of time (2, 4, 8, 12, and 24 h). The hydrolysis results were compared by scanning electron microscope (SEM), X‐ray power diffractometer (XRD), and differential scanning calorimetry (DSC). The SEM results revealed that both of the PLO and PTB starches showed the same hydrolysis mechanism, which indicated that the glucoamylase primarily attacking the exterior of starch granules and then the interior. The results of XRD revealed the crystalline type of PTB starch changed from C‐type to A‐type with crystallinity reducing from 43.5 to 20.9% during the hydrolysis. Unlike PTB starch, the PLO starch did not show marked changes in crystalline style but lower degree of crystallinity was obtained from 32.4 to 13.7% during the hydrolysis. All the XRD results demonstrated that B‐type polymorph was preferentially degraded than A‐type polymorph in the C‐type starch. The DSC results revealed that both of the PLO and PTB starches showed decreased enthalpy of gelatinization (ΔH_gel) and gelatinization temperature range (R)‐value after hydrolysis, while the gelatinization temperature (T_p) indicated different tendency, initially ranging from 68.6 to 64.3°C and then increasing to 67.8°C for PLO starch. While for PTB starch, the T_p‐value showed progressive reduction from 85.4 to 74.3°C during the whole process.     

Hydrolysis of Tropical Tuber Starches by Bacterial and Pancreatic α-Amylases

Action of porcine pancreatic and Bacillus subtilis α-amylases on native tuber starches of yam (Dioscorea alata), sweet potato (Ipomea batatas) and tannia (Xanthosoma sagittifolium) was studied in comparison with the well known potato and cassava starches. Large differences in enzymes susceptibilities were observed when studied on 24h. Yam starch was 3.5% hydrolysed with 2,8 μkat amylase/g starch, three times less than potato and tannia starches while sweet potato starch was 53% hydrolysed, two times less than cassava starch. Except yam, level of hydrolysis was higher with porcine pancreatic amylase than with the Bacillus subtilis amylase while initial hydrolysis rate was lesser. Microscopic observations and image analysis pointed out that the polyhedric shaped granules of tannia, sweet potato and cassava starches were much more damaged than the spherical ones. Pitting occured preferentially on the edges of the granules and the enzymes penetrated into the starch granule by pores and canals of corrosion. Conversely to other starches, hydrolysis of yam starches evidenced greater differences between action of Bacillus subtilis and pancreatic α-amylases. The enzymes acted by pitting some parts of the granules surface, the number of pores and their size being related to enzyme source.     

Comparison of the morphological,crystalline, and thermal properties of different crystalline types of starches after acid hydrolysis

A‐type maize starch, B‐type Fritillaria ussurensis, and C‐type Rhizoma dioscorea starches were hydrolyzed (32 days) with 2.2 N HCl. Regardless of the crystallinity level, starch with predominant B‐crystalline type was less susceptible to acid degradation than A‐type and C‐type starches, and initial rates of hydrolysis in B‐type was lower than others. The SEM and XRD results revealed that different types of starch displayed different hydrolysis mechanisms. The acid corrosion started from the exterior surface of A‐type and B‐type starches followed by the core of granules. However, the hydrogen ions primarily attacked the interior of the C‐type R. dioscorea starch granules and then the exterior. FT‐IR results confirmed that the amorphous regions in the starch granules were hydrolysed first. After 8–32 days of hydrolysis, the acid‐modified C‐type starch showed typical A‐type characteristics upon analysis of the XRD pattern. The average particle size of hydrolytic starch decreased with increasing hydrolysis time. The thermal results revealed that the hydrolytic starch showed lower ΔH than the native starch, while displaying higher peak width (T_c − T_o) value.     

Physicochemical Characterization and Application of Yam (Dioscorea cayenensis‐rotundata) Starch as a Pharmaceutical Excipient

Yam starch from Dioscorea cayenensis‐rotundata complex was isolated and characterized by scanning electron microscopy (SEM), particle size analysis, X‐ray diffraction, differential scanning calorimetry (DSC), compaction and rheology, and compared to maize (Zea mays) and potato (Solanum tuberosum) starches. Yam starch exhibited a log‐normal distribution of flattened ovoid shaped granules with a mean particle size of 25 µm. X‐ray diffraction showed a C‐type crystalline pattern with the degree of relative crystallinity estimated to be 34%. DSC analysis suggests that the crystalline regions in yam starch are thermally and structurally more stable as in maize and potato. Irrespective of the relative humidity (39, 67, 78% R.H.) yam starch exhibited higher moisture uptakes than maize starch and lower than potato. Intermediate values of swelling power and amylose leaching were obtained for yam as compared to maize and potato. Compaction properties of yam and potato starches were similar. However, compacts from yam presented a relatively lower tensile strength. Aqueous starch systems (4%) of yam and maize starches showed analogous shear‐thinning (pseudo‐plastic) behavior suitably described by the power‐law model. These results support the potential use of yam starch as excipient comparable to potato starch in pharmaceutical solid forms and as thickening agent similar to maize in pharmaceutical applications.     

Characterization of Residues from Partially Hydrolyzed Potato and High Amylose Corn Starches by Pancreatic α‐Amylase

High amylose corn starch (HACS) and potato starch were hydrolyzed by pancreatic α‐amylase in vitro. Residues after hydrolysis were collected and characterized for their physicochemical properties and molecular structure. Compared with raw starches, residues had lower apparent amylose contents and higher resistant starch contents. The gelatinization enthalpy of residues from HACS increased while enthalpy of residues from potato starch decreased from 15.4 to 11.3 J/g. Peak viscosity and breakdown values of the residues from potato starch were markedly decreased but final viscosity values did not show much change. Chain length distribution of debranched amylopectin from the residues indicated that the relative portion of short chain in the residue decreased for both starches. More molecules with intermediate chain length (DP 16—31) were found in residue after 48‐h hydrolysis of potato starch.     

Response surface optimization and characteristics of Indica rice starch‐based fat substitute prepared by α‐amylase

Response surface methodology (RSM) was used to study the effect of enzyme to substrate ratio (11.8–23.6 U α‐amylase/g rice starch), hydrolysis temperature (90–100°C) and pH value (6.0–6.6) on the gel strength of rice starches‐based fat substitute using α‐amylase hydrolysis. The optimum conditions obtained from response surface analysis was 16.52 U/g enzyme dosage, 92°C hydrolysis temperature while the influence of pH was found insignificant in the range tested. Under these optimum conditions, the gel strength of this fat substitute was 113 g/cm², very close to the gel strength of butter of 114 g/cm², while the solubility of the substitute was 1.33 ± 0.01% and the swelling power 4.85 ± 0.02. There were no observable differences in the granular size distribution between the untreated rice starch and the hydrolyzed rice starch. Rheological properties of this rice starch‐based fat substitute implied that it is easier for the substitute to form three‐dimensional networks under 34°C.     

Influence of Selected Parameters of Starch Gelatinization and Hydrolysis on Stability of Amylose‐Lipid Complexes

Dissociation of amylose – lipid complexes (AMLs) upon gelatinization and enzymatic hydrolysis of wheat starch has been examined using differential scanning calorimetry (DSC). Digestion of starch by the thermostable α‐amylase THERMAMYL 120L was carried out under conditions applied for enzymatic hydrolysis of wheat starch in industry, i.e. 5 min incubation at 105°C, followed by 60–90 min hydrolysis at 95°C. For comparison reasons, samples of wheat starch slurries were incubated under the same conditions but in the absence of enzyme. The enthalpy and temperature of AMLs dissociation and the shapes of peaks in the DSC endo‐ and exotherms depended on conditions of starch processing prior to DSC measurements. Wheat starch gelatinization coupled with its digestion by α‐amylase resulted in more pronounced AML degradation as compared to gelatinization in the absence of enzyme. Different shapes of peaks in thermograms and different temperatures of AMLs dissociation and reassociation indicate that different AML polymorphs were generated in the examined samples. Their concentrations depended on conditions (temperature, time and the presence or absence of α‐amylase) of wheat starch treatment.     

Isolation of a Recombinant Bacillus sp. TS‐23 α‐Amylase by Adsorption‐Elution on Raw Starch

A Bacillus sp. TS‐23 α‐amylase produced by recombinant Escherichia coli was adsorbed onto raw starch and the adsorbed enzyme was eluted with maltose or maltodextrin in 50 mM Tris/HCl buffer (pH 8.5). The adsorption‐elution procedure resulted in a yield of 53% α‐amylase activity and sodium dodecyl sulfate‐polyacrylamide gel electrophoresis (SDS/PAGE) analysis showed that the eluted α‐amylase had a molecular mass of approximately 64 kDa. Raw starch could be used repeatedly in the adsorption‐ elution cycle with good reproducibility. Scanning electron microscopy of the isolated corn starch exhibited a smooth appearance of the granules before adsorption and only a small change in appearance after three adsorption‐elution cycles. These results suggest that the raw starch adsorption‐elution technique has a great potential in the isolation of Bacillus sp. TS‐23 α‐amylase from the culture broth of recombinant E. coli.     

Preparation and Characterization of Annealed‐Enzymatically Hydrolyzed Tapioca Starch and the Utilization in Tableting

Tapioca starch was annealed at 60°C for 90 min followed by hydrolysis with α‐amylase at 60°C at various lengths of time (30, 60 and 120 min) to obtain high‐crystalline starches. The reaction products were subjected to spray drying to obtain annealed–enzymatically hydrolyzed–spray dried tapioca starch (SANET) in the form of spherical agglomerated granules. The properties of SANET were compared with those of annealed–spray dried tapioca starch without enzymatic treatment (SANT) and native–spray dried tapioca starch (SNT). Scanning electron micrographs of the starch samples were used to study the morphological changes and to suggest the mode of enzyme attack during hydrolysis. The á‐amylase preferentially attacked the interior of the starch granules, leaving a deep round hole on the starch granule surface. It was found by X‐ray diffraction that both annealing and amylolysis did not alter the A type diffraction pattern. The% relative crystallinity of SANET was raised with increasing hydrolysis time and with decreasing amylose content. High performance size exclusion chromatography (HPSEC) demonstrated the decrease of the degree of polymerization (DP) of the amylose fraction of SANET after prolonged hydrolysis. For the utilization of SANET as tablet filler, it was directly compressed by a tablet compression machine at 4 kN to obtain tablets. The increased relative crystallinity of starch resulted in increased crushing strength and disintegration time, but in a decreased tablet friability.     

Influence of α‐amylolysis on the formation of electron density inhomogeneities on the surface of starch granules

Low‐temperature nitrogen adsorption and SAXS were used to analyze the granule surface of wheat and rice starches before and after an action of Bacillus subtilis α‐amylase. Results obtained by the first method showed that α‐amylolysis caused surface changes arising of new pores or enlarging diameters of pores already existing. According to the SAXS results, the action of enzymes caused complete sharpening of a solid skeletal boundary in the case of wheat starch granules. Partial sharpening of this boundary in the case of rice starch granules could be attributed to the relatively high content of proteins in rice starch.     

Enhanced production of α‐amylase from Bacillus subtilis subsp. spizizenii in solid state fermentation by response surface methodology and its evaluation in the hydrolysis of raw potato starch

This is an attempt to lower the cost of starch hydrolysis by the discovery of new generation α‐amylase. A natural isolate of Bacillus subtilis subsp. spizizenii was capable of producing appreciable amounts of raw potato starch digesting α‐amylase in solid state fermentation of wheat bran. The enzyme productivity has been substantially enhanced by supplementing various nutrients and statistically studying their interactions by response surface methodology. A central composite design for amylase production system elucidated a wheat bran‐based medium supplemented with soybean meal, threonine, and B‐complex vitamins predicting a yield of 521 391 U/g dry solids. The enzyme preparation could effectively digest 5–15% suspension of insoluble potato starch in 6 h revealing the dextrose equivalent of 32–44. The supplementation of a glucoamylase preparation, thereafter, brought about complete saccharification. The yield achieved in the statistically optimized amylase system may be one of the best to date and its capability in directly liquefying raw potato starch granules makes this study novel.     

Effect of Heat‐Moisture Treatment on Structural and Thermal Properties of Rice Starches Differing in Amylose Content

Starches from glutinous rice (1.4% amylose), Jasmine rice (15.0% amylose) and Chiang rice (20.2% amylose) were exposed to heat‐moisture treatment (HMT) at 100 °C for 16 h and at different moisture levels (18, 21, 24 and 27%). The effect of heat‐moisture treatment on structural and thermal properties of these three rice starches was investigated. The HMT did not change the size, shape and surface characteristics of rice starch granules. The A‐type crystalline pattern of rice starches remained unchanged after HMT. The relative crystallinity (RC) and the ratio of short‐range molecular order to amorphous (RSA) of heat‐moisture treated glutinous and Jasmine rice starches decreased with increasing moisture level of the treatments. In contrast, the RC of the treated Chiang rice starch remained practically unchanged. A peak of crystalline V‐amylose‐lipid complexes was clearly presented in all treated Chiang rice starches. The peak became progressively stronger with increasing moisture level of the treatment. Differential scanning calorimetry (DSC) of all treated rice starches showed a shift of the gelatinization temperature to higher values. Increasing moisture level of the treatments increased the onset gelatinization temperature (T_o) but decreased the gelatinization enthalpy (ΔH) of rice starches. A broad gelatinization temperature range (T_c‐T_o) with a biphasic endotherm was found for all treated Chiang rice starches and Jasmine rice starch after HMT₂₇ (HMT at 27% moisture level). Additionally the (T_c‐T_o) of treated Chiang rice starches increased linearly with increasing moisture level of the treatments.     

Suppression of Fusarium graminearium growth by differently structured starch types

We investigated the growth behavior and amylolytic enzymes of Fusarium graminearum cultivated on different types of native starch granules including barley (A‐type crystalline polymorph), potato and Curcuma zedoaria (B‐type crystalline polymorph), cassava (C‐type crystalline polymorph), and high AM maize (A + Vh‐type crystalline polymorphs). F. graminearum grew poorly on B‐type starches and the accumulation of biomass was similar to that obtained for fungi cultivated under carbohydrate starvation conditions. In comparison, three‐ to fivefold higher accumulation of fungal biomass was observed for growth on the A‐, C‐ and A + Vh‐type starches. Fungal glucoamylase and α‐amylase activity increased over time in the presence of native starch granules. Interestingly, resistant B‐type starches induced the highest amylolytic activity indicating that F. graminearum interacts with B‐type granules although only limited degradation occur. Starch degradation products maltose and malto‐oligosacharides was found to increase glucoamylase and α‐amylase activity, whereas glucose acted as a catabolite repressor.     

In vitro Digestibility of Raw Starches Extracted from five Yam (Dioscorea spp.) Species Grown in Jamaica

Starch granules from Round leaf yellow yam, Negro yam, Sweet yam, Bitter yam and Chinese yam grown in Jamaica were isolated and characterized. The amylose content, granular size, crystallinity, and digestibility by α‐amylase were determined. The granules obtained were of three crystalline types. Round leaf yellow yam, Negro yam and Sweet yam were found to be type‐B, while Chinese yam and Bitter yam were type‐C and type‐A, respectively. Round leaf yellow yam had the highest amylose content (26.5%) while Chinese yam had the lowest (11.1%). The granule size varied between 1–3 μm for Chinese yam and 16–42 μm for Round leaf yellow yam. Significant variations in digestibility of the granules were observed. Raw starches from Chinese yam and Bitter yam were the most susceptible to α‐amylase digestion (porcine pancreatic α‐amylase, pH 5.5, 0.02% CaCl₂, 40°C, 24 h) with 21.27 ± 0.01% and 18.11 ± 0.02% degradation, respectively, while Round leaf yellow yam, Negro yam and Sweet yam starches were the least susceptible, with 13.74 ± 0.03%, 14.98 ± 0.08%, and 15.32 ± 0.04% enzymatic degradation, respectively.     

Effect of acid–ethanol on the physicochemical properties of Dioscorea opposita Thunb. and Pueraria thomsonii Benth. starches

Dioscorea opposita Thunb. and Pueraria thomsonii Benth. starches were hydrolyzed with 0.36% HCl under ethanol conditions during 1, 3, 5, 8, and 12 h. Structural and physicochemical characteristics studies of C‐type D. opposita Thunb. and P. thomsonii Benth. starches were carried out using SEM, XRD, and FTIR spectroscopy. From the FTIR result, it was determined that the amorphous areas of starch granules were hydrolyzed successfully. The SEM and XRD results revealed that the interior was preferentially hydrolyzed and the degree of crystallinity increased for D. opposita and P. thomsonii starches. However, the two kinds of starches were hydrolyzed in different ways. The starch granules of D. opposita presented bread‐like in shape after acid hydrolysis. While the P. thomsonii Benth. starch granules were degraded for concaves formed on the surface, and several small particles appeared. Water‐binding capacity and AM content were observed to be lower for both starches. The thermal results revealed that the hydrolyzed starches showed the lower ΔH_gel than the native starches, which can be related to the preferential destruction of amorphous areas in the granules.     

Acid Diffusion into Rice Boluses is Influenced by Rice Type,Variety, and Presence of α‐Amylase

Breakdown of rice during gastric digestion may be influenced by rice structure, presence of salivary α‐amylase, and hydrolysis by gastric acid. During mastication, saliva is mixed with rice, allowing α‐amylase to begin starch hydrolysis. This hydrolysis may continue in the gastric environment depending on the rate at which gastric acid penetrates into the rice bolus. The objective of this study was to determine the acid uptake into rice boluses with and without α‐amylase in saliva. Two types each of brown and white rice (medium and long grain), were formed into a cylindrical‐shaped bolus. Each bolus was sealed on all sides except one to allow one‐dimensional mass transfer, and incubated by immersion in simulated gastric juice at 37 °C under static conditions. Acidity of the boluses was measured by titration after 1 to 96 h of incubation. Effective diffusivity of the gastric juice through the bolus was estimated using MATLAB. Average acidity values ranged from 0.04 mg HCl/g dry matter (medium grain white rice, no incubation) to 10.01 mg HCl/g dry matter (long‐grain brown rice, 72 h incubation). The rice type, presence of α‐amylase, and incubation time all significantly influenced rice bolus acidity (P < 0.001). Effective diffusivity of gastric juice into the bolus was greater in brown rice than in white rice. These results indicate that starch hydrolysis by α‐amylase may continue in the stomach before the gastric acid penetrates the rice bolus, and the rate of acid uptake will depend on the type of rice consumed.     

山药及零余子酸、酶处理淀粉的理化性质

利用酸和淀粉酶、糖化酶分别处理山药及零余子淀粉,利用扫描电镜、X射线衍射及流变仪等测定不同处理组淀粉的理化特性。结果表明：山药和零余子原淀粉颗粒较为完整,没有裂缝,呈卵圆形,表面光滑;酶、酸处理均会导致淀粉降解,酸处理淀粉表面破坏最为严重;零余子原淀粉、酸处理和酶处理的淀粉结晶度分别为32.02%、27.02%和31.13%,而山药原淀粉、酸处理和酶处理淀粉结晶度分别为34.32%、29.27%、33.55%。淀粉糊透光率、凝沉百分比表明零余子原淀粉最易老化,流变特性表明不同处理组的淀粉均具有弱凝胶特性。     

Effect of Enzymatic Hydrolysis of Wheat Starch on Amylose‐Lipid Complexes Stability

In the process of enzymatic hydrolysis of wheat starch to glucose, the presence of amylose‐lipid complexes (AML's) decreases swelling and dissolving capacity and the water binding capacity of starch, thus impeding the access of amylolytic enzymes to the starch granules. The aim of the work was to define the relationship between the stability of AML's and the conditions of the enzymatic hydrolysis of wheat starch such as the kind of enzymatic preparation (only amylolytic or both amylolytic and lipolytic) and time of hydrolysis. Hydrolysates were produced from wheat starch liquefied with bacterial α‐amylase BAN 240L, subjected to further treatment with the enzymatic preparation Spezyme GA 300W, containing glucoamylase and lysophospholipase and, for comparison, only with a glucoamylase preparation (AMG 300L). The effect of amylolytic and lipolytic enzymes on the stability of AML's in the process of wheat starch hydrolysis was estimated. The amount of fatty acids released during hydrolysis was determined with gas‐liquid chromatography (GLC) and by differential scanning calorimetry (DSC) measuring the enthalpy of decomposition of AML's. The investigations revealed a differentiated effect of the individual enzymatic preparations on the degree of degradation of AML's. Amylose‐lipid complexes were more susceptible to the attack of preparation Spezyme GA 300W than to the digestion by α‐amylase BAN 240 L and glucoamylase AMG 300L.