Characteristics of native and enzymatically hydrolyzed Zea mays L., Fritillaria ussuriensis Maxim. and Dioscorea opposita Thunb. starches

Comparative studies on glucoamylase hydrolysis of A-type Zea mays L., B-type F. ussuriensis Maxim., and C-type Dioscorea opposita Thunb. were carried out by scanning electron microscope (SEM), X-ray diffractometer (XRD), Fourier transform infrared (FT-IR) spectra and differential scanning calorimetry (DSC). Maize, Fritillaria, Dioscorea starches were hydrolyzed with glucoamylase for 2, 4, 8, 12 and 24 h, respectively. The SEM and XRD results revealed that A-type, B-type starch and C-type starch displayed different hydrolysis mechanisms. A-type starch was digested with enzyme penetrating into starch granules through natural pores on the surface and disrupted the interior of the starch granules. The glucoamylase worked by attacking the surface of B-type starch and forming cracks. When endo-corrosion occurred, the internal part of the granule was corroded through small cracks. However, the glucoamylase primarily attacked the interior of the C-type starch granules and then the exterior of starch granules. FT-IR confirmed that the amorphous regions in the starch granules are firstly hydrolyzed and could be hydrolyzed completely as long as the hydrolysis time is sufficient. The transition temperatures and enthalpy of gelatinization (ΔH_gel) were determined using DSC. According to the gelatinization parameters, it could be further proved amorphous and crystalline structures were hydrolyzed.     

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.     

Morphology and structural characterization of high-amylose rice starch residues hydrolyzed by porcine pancreatic α-amylase

High-amylose starches are attracting considerable attention because of their potential health benefits and industrial uses. Enzyme hydrolysis of starch is involved in many biological and industrial processes. In this paper, starches were isolated from mature grains of high-amylose transgenic rice line (TRS) and its wild type rice cultivar Te-qing (TQ). The morphological and structural changes of starch residues following porcine pancreatic α-amylase (PPA) hydrolysis were characterized by SEM, DSC, XRD, ¹³C CP/MAS NMR, and ATR-FTIR. TQ starch was hydrolyzed faster than TRS starch. PPA pitted the starch granule surface first, then penetrated into the interior and hydrolyzed the granule from the inside out. The peripheral region of subgranule and the surrounding band of TRS starch were more resistant to PPA hydrolysis than the external region of TQ starch. Both amorphous and crystalline (the long- and short-range ordered) structures were simultaneously hydrolyzed in TQ starch. A-type polymorph of TRS C-type starch was hydrolyzed faster than B-type polymorph. The long-range ordered structure was hydrolyzed faster than the amorphous structure, whereas the short-range ordered structure was hydrolyzed slower than the amorphous structure in TRS starch in the initial hydrolysis stage. The above results indicated that B-type crystallinity and short-range ordered structure in the peripheral region of subgranule and the surrounding band of TRS starch increased the resistance of TRS starch to PPA hydrolysis.     

Granular structure and allomorph position in C-type Chinese yam starch granule revealed by SEM,C CP/MAS NMR and XRD

The crystalline microstructure and polymorphism of C-type starch from Chinese yam were evaluated by scanning electron microscope (SEM), ¹³C cross-polarization magic-angle spinning NMR (¹³C CP/MAS NMR) and powder X-ray diffraction (XRD) technique and acid hydrolysis method. Morphological changes during acid hydrolysis showed that the amorphous or the less crystalline areas were essentially located at the center part of C-type starch granules whereas the semi-crystalline and amorphous growth rings were found mainly in the outer part of the granules. ¹³C CP/MAS NMR and XRD results revealed that B-type allomorph was hydrolyzed more rapidly than A-type one. The amorphous or less crystalline areas were predominantly composed of B-allomorph whereas the outer semi-crystalline and amorphous growth rings were mostly composed of A-type allomorph. The A- and B-type allomorph coexisted in the individual C-type starch granule. B-type allomorph basically existed at the center part of the granules which was surrounded by the A-type allomorph in the peripheral part of granules.     

The structure of C-type Rhizoma Dioscorea starch granule revealed by acid hydrolysis method

Scanning electron microscope (SEM), X-ray powder diffraction (XRD) and cross polarisation/magic-angle spinning (CP/MAS) ¹³C nuclear magic resonance (NMR) have been used for the structural characterisation of C-type starch granule during acid hydrolysis. SEM shows that the amorphous areas mainly locate the core part of C-type starch granules, while the crystalline areas mainly exist in the peripheral region of starch granules. XRD analysis reveals that the B-type polymorph present in the C-type starch granule are preferentially degraded or degraded faster than the A-type polymorph. NMR spectra confirm that the amorphous regions in the starch granules are firstly hydrolysed and could be hydrolysed completely as long as the hydrolysis time is sufficient. After 40 days of hydrolysis, the acid-modified starch shows typical A-type characteristics upon analysis of the XRD pattern or the ¹³C CP/MAS NMR spectra.     

Conformation and location of amorphous and semi-crystalline regions in C-type starch granules revealed by SEM,NMR and XRD

The conformations and locations of amorphous and semi-crystalline regions in C-type starch granules from Chinese yam were evaluated by a combination of morphology and spectroscopy studies during acid hydrolysis. Scanning electron micrographs showed that amorphous or less crystalline areas were essentially located in the centre part of C-type starch granules, whereas the semi-crystalline and amorphous growth rings were found mainly in the outer part of the granules. ¹³C cross-polarization magic angle spinning NMR (¹³C CP/MAS NMR) showed that amorphous regions were hydrolyzed faster than the crystalline ones. In addition, B-type polymorphs were shown to be hydrolyzed more rapidly than A-types. Powder X-ray diffraction (XRD) also revealed that the B-polymorph was hydrolyzed more rapidly than the A-type. XRD showed that the amorphous or less crystalline areas were mainly located in the core of starch granules, while the amorphous growth rings are distributed toward the outside of the granules and alternatively arranged with semi-crystalline growth rings. The amorphous or less crystalline areas predominantly consisted of the B-polymorph whereas the outer semi-crystalline and amorphous growth rings were mostly composed of the A-polymorph.     

Morphology and structural properties of high-amylose rice starch residues hydrolysed by amyloglucosidase

High-amylose starches are attracting considerable attention because of their potential health benefits and industrial uses. Enzyme hydrolysis of starch is involved in many biological and industrial processes. In this paper, starches were isolated from high-amylose transgenic rice (TRS) and its wild type rice, Te-qing (TQ). The morphological and structural changes of starch residues following Aspergillus niger amyloglucosidase (AAG) hydrolysis were investigated. AAG hydrolysed TQ starch from the granule surface, and TRS starch from the granule interior. During AAG hydrolysis, the content of amorphous structure increased, the contents of ordered structure and single helix decreased, and gelatinisation enthalpy decreased in TQ and TRS starch residues. The A-type polymorph of TRS C-type starch was hydrolysed faster than the B-type polymorph. The short-range ordered structure and B-type polymorph in the peripheral region of the subgranule and the surrounding band of TRS starch increased the resistance of TRS starch to AAG hydrolysis.     

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.     

The effect of acid and glucoamylase hydrolysis on partial physicochemical properties and characteristics of Dioscorea alata Linn. starch

Dioscorea alata Linn. (D. AL) rich in starch was widely used as food and medicine. This work aimed to compare the effect of acid and glucoamylase on the starch of different reaction time. The physicochemical properties of the native and hydrolyzed starches, such as swelling power (SP), starch solubility (SOL), and paste clarity were also determined in this study. The native and modified starches by acid and glucoamylase were compared and characterized by scanning electron microscope (SEM) and secondary derivative FT‐IR spectroscopy. The morphological characteristics of native D. AL starch granules revealed by SEM varied from round to oval or elliptic with smooth surface. SEM showed that glucoamylase primarily attacked the exterior of starch granules and permeate into the interior by the cracks formed on the surface. Owing to acid hydrolysis, the starch granules were depressed and transformed to bread‐like in shape and then broke into irregular lumps with particles adhered. From secondary derivative FT‐IR spectrum, the intensities of the two peaks from acid and enzyme hydrolysis starches at 1137 and 1066/cm were stronger than native starch. Some new peaks appeared due to hydrolysis. The digested starches exhibited a lower SOL and SP than native starch in water as the temperature increasing. The light transmittance of the digested starches was significantly higher than that of native starch paste and decreased with increase in storage period.     

Properties of Residual Starches of Sugary-2 and Sugary-2 Opaque-2 Maize Following Amylase Hydrolysis

Properties of residual starches of sugary-2 opaque-2 and sugary-2 maize starch granules hydrolyzed with glucoamylases were investigated. A crude and two crystalline glucoamylases were used. The amylopectin fractions of both starches hydrolyzed easier than that of amylose with all enzymes. Residual starches hydrolyzed by the crude glucoamylase accumulated low-molecular weight materials, which was not observed in residual starches attacked by crystalline glucoamylase. It was suggested that in the crude enzyme the contaminating α-amylase caused the accumulation of the minified fraction. It is also suggested that the crystalline region of sugary-2 opaque-2 starch may consist of a mixture of A-type and B-type patterns. Evidence for this was from observation of the changes in X-ray diffraction patterns of residual starch following amylase and acid hydrolysis.     

体内外消化高直链抗性水稻淀粉的波谱特性

以基因工程培育出来的高直链抗性淀粉水稻(TRS)为材料,利用粉末X射线衍射、傅里叶变换红外光谱分析技术,研究体内外消化TRS淀粉的波谱特性。结果表明：不被消化的抗性淀粉直链淀粉含量显著升高;TRS淀粉为C型晶体淀粉,由A型和B型晶体淀粉组成,其中A型晶体比B型晶体消化降解快,B型晶体具有抗消化降解的作用;TRS淀粉的无定形区域易被消化降解。     

Crystallography,morphology and thermal properties of starches from four different medicinal plants of Fritillaria species

To fully understand the medicinal plant, Fritillaria, and its species, we investigated the physical properties of starch contained in four Fritillaria species, Fritillaria thunbergii Miq., Fritillaria ussurensis Maxim., Fritillaria pallidifloca Schrenk and Fritillaria cirrhosa D.Don, by means of various analytical methods. The crystal type of the former three kinds of Fritillaria starches was in characteristic B-type, which was in agreement with the crystal type of potato starch. However, the cirrhosa F. starch showed a typical C_B-type pattern. The degrees of crystallining of the four Fritillaria starches were about 43.2%, 40.5%, 44.8% and 41.8%, corresponding to thunbergii F. starch, ussurensis F. starch, pallidifloca F. starch and cirrhosa F. starch. The granule sizes of the former two Fritillaria starches ranged from 5 to 40 μm, and were cycloidal or elliptic-shaped. However, the latter two Fritillaria starch granules had granule sizes ranging from 5 to 50 μm, and the granule shape varied from oval to irregular or cuboidal. From the thermogravimetric analysis, it was concluded that the thermal stabilities of the four kinds of starch differed from each other, due to their different structures.     

Characterization of nanoparticles prepared by acid hydrolysis of various starches

Various starches of different AM contents and origins such as wx maize, normal maize, high AM maize, potato, and mungbean starches were hydrolyzed using a H₂SO₄ solution (3.16 M) at 40°C for 7 days, and the starch particles were isolated from the hydrolysates by centrifugation. The hydrolysis rates varied from 61.4 to 90.9% depending on the starch type. Unexpectedly, A‐type starches were more resistant to the acid hydrolysis than B‐type starches. XRD results revealed that the starch particles with B‐crystalline type exhibited a decrease in peak intensity. In addition, in a DSC analysis, the crystals remaining in the B‐type starch particles were readily disrupted in the water dispersion so that no melting endotherm appeared. Electron microscopy confirmed that the starch particles had round or oval shapes with diameters ranging from 40 to 70 nm, which possibly represented the starch blocklets in granules. The acid degraded mainly AM and long AP chains, resulting in increasing the proportion of short chains.     

醇酸降解淀粉制备糊精的研究

采用醇酸水解大米淀粉、高直链玉米淀粉和木薯淀粉,发现：在相同条件下,正丁醇-盐酸水解的木薯淀粉溶解性最大,且在水解3 d后无显著增加。因此,从颗粒结构、晶体结构和分子结构等方面系统地研究醇酸降解木薯淀粉制备糊精过程。经正丁醇-盐酸水解后,木薯淀粉颗粒破坏程度严重,且随水解时间的增加,大部分木薯淀粉颗粒均失去了原有的颗粒形状;经正丁醇-盐酸水解7 d后,木薯淀粉的粒径下降了42.0%。X-射线衍射结果表明正丁醇-盐酸水解木薯淀粉在2θ为20°处出现了衍射峰,证明淀粉在正丁醇-盐酸水解过程中形成了直链淀粉-正丁醇络合物。高效分子排阻色谱和淀粉-碘全波长扫描分析表明：木薯淀粉经过正丁醇-盐酸水解后分子量在4 h内急剧下降,水解4 h后下降趋势减缓,水解3 d后分子量无显著变化并趋于稳定。上述结果表明,采用正丁醇-盐酸水解木薯淀粉3 d为制备糊精的最佳条件。     

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.     

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.     

Morphology and structure of chestnut starch isolated by alkali and enzymatic methods

The structure and morphology of starch from fruits of two chestnut (Castanea sativa Mill.) varieties, Martainha and Longal, isolated by alkaline (A3S) and enzymatic (ENZ) methods were assessed. Chestnut starch granules were found to be round and oval in shape, consisting of medium/small granules, with a mean granule size ranging between 9 and 13 μm. Isolated chestnut starch appeared to the naked eye as a white powder, with high values of L^∗, and the Longal variety produce starches duller than Martainha. No differences between samples were observed by FTIR analysis. The X-ray patterns of isolated starches are of C-type (more specifically of C_b type) with a relative crystallinity between 31.5% and 39.8%. The ¹³C CP/MAS NMR spectra are similar for both varieties but different for the used isolation methods. The amorphous phase in the starch granules isolated by A3S methods was lower than that of the starch extracted by the ENZ method, making the B-type allomorph in the C-type starch granules more evident than in the A-type. Those differences in the structure of isolated starches are shown by a lower degree of damage, and a higher level of crystallinity of starches isolated by the A3S method, which means that its original structure is less affected or partially destroyed. This study would be helpful to better understand the relationships among structure and functional properties for a eventual industrial application of chestnut starches.     

湿热处理对不同晶型淀粉理化性质及消化性的影响

采用三种不同晶型淀粉即玉米淀粉(A型)、马铃薯淀粉(B型)、豌豆淀粉(C型)为原料,在水分含量为25%、温度120℃条件下湿热处理13 h,研究湿热处理对不同晶型淀粉的理化性质及消化性的影响。研究表明,与原淀粉相比,经湿热处理的三种淀粉的结晶结构均发生了改变,玉米淀粉由A型变为了A+V型,马铃薯淀粉和豌豆淀粉分别由B型和C型变为了A型;三种淀粉颗粒表面均出现了不同程度的破损;三种淀粉的部分颗粒的偏光十字的中心强度有所减弱;三种淀粉样品的糊化温度均升高,但A型和B型淀粉的焓值降低,而C型淀粉的焓值升高;三种淀粉的抗性组分含量均有所升高,抗消化性显著增强,其中C型淀粉变化最明显。     

Physicochemical Property of Huaishan (Rhizoma Dioscorea) and Matai (Eleocharis dulcis) Starches

The physicochemical properties of starch, the main component in tuberous root of Huaishan (Rhizoma Dioscorea) and corm of Matai (Eleocharis dulcis) was investigated and compared with those of a Japanese yam, “Yamanoimo” starch, and potato starch. Mean particle sizes of Huaishan and Matai starches were 24 ± 5 μm and 12 ± 5 μm, respectively. X-Ray diffraction pattern suggests that Huaishan starch was B-type or C-type just close to B-type and Matai starch was A-type or C-type just close to A-type. Apparently, the intermediate component (IntCom) of Huaishan starch, which was obtained by fractionation of the starch into amylose and amylopectin, may still contain amylose and amylopectin. IntCom of Matai starch has an intermediate nature between amylose and amylopectin. It is concluded that the amylopectin molecules of Huaishan starch contain a larger amount of longer branch-ed chains and those of Matai starch contain a larger amount of shorter branched chains. Amylograms of Matai and Huaishan starches suggest that the gelatinized starches are difficult to retrograde. Digestibility of Huaishan starch by an α-amylase was the highest among the tested starches.     

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.