Comparison of potato amylopectin starches and potato starches — influence of year and variety

Starches from three potato varieties and their respective transformants producing amylopectin starch were studied over a period of 3 years. The gelatinisation, swelling and dispersion properties were studied using differential scanning calorimetry (DSC), X-ray diffraction, swelling capacity measurements and a Brabender Viscograph.

The potato amylopectin starches (PAP) exhibited higher endothermic temperatures as well as higher enthalpies than the normal potato starches (NPS). PAP samples gave rise to an exceptionally sharp viscosity peak during gelatinisation and a relatively low increase in viscosity on cooling. Swelling capacity measurements showed that PAP granules swelled more rapidly, and that the dispersion of the swollen granules occurred at a lower temperature (85°C). Analysis of variance (ANOVA) also revealed that the year influenced the DSC results, and that both year and variety affect some of the Brabender parameters. Furthermore, the PAP and NPS samples were subjected to heat–moisture treatment at three different moisture levels, and the Brabender viscosity properties were studied.     

On the effect of surface active agents and their structure on the temperature-induced changes of normal and waxy wheat starch in aqueous suspension. Part I. Pasting and calorimetric studies

Pasting and calorimetric studies of normal and waxy wheat starch were performed in the presence of a series of ionic (sulphates, trimethyl ammonium bromides) and non-ionic (monoglycerides, maltosides) short (12 carbon atoms) and long (16 carbon atoms) n-alkyl chain surfactants. With the exception of the alkyl ammonium bromides, all of the short chain surfactants lower the pasting temperature (PT) in normal wheat starch, while the long chain surfactants have the opposite effect. Contrary, regardless of their chain length, all ionic surfactants lower the PT in waxy wheat starch while the non-ionic surfactants induce small, sometimes almost negligible changes in the PT. Calorimetric studies revealed the absence of a direct connection between the effect of surfactants on the onset of the starch gelatinization transition and the PT. However, in the presence of all surfactants, except the alkyl ammonium bromides, the PT of normal wheat starch was found to lie within or very close the temperature range within which the dissociation of the amylose–surfactant complexes takes place. Waxy wheat starch, in contrast, pasted at temperatures that fell within the temperature range of the starch gelatinization transition. This is taken as evidence of the existence of a correlation between the PT and the dissociation of the amylose–surfactant complexes.     

Accumulation of multiple-repeat starch-binding domains (SBD2–SBD5) does not reduce amylose content of potato starch granules

This study investigates whether it is possible to produce an amylose-free potato starch by displacing the amylose enzyme, granule-bound starch synthase I (GBSSI), from the starch granule by engineered, high-affinity, multiple-repeat family 20 starch-binding domains (SBD2, SBD3, SBD4, and SBD5). The constructs were introduced in the amylose-containing potato cultivar (cv. Kardal), and the starches of the resulting transformants were compared with those of SBD2-expressing amylose-free (amf) potato clones. It is shown that a correctly sized protein accumulated in the starch granules of the various transformants. The amount of SBD accumulated in starch increased progressively from SBD to SBD3; however, it seemed as if less SBD4 and SBD5 was accumulated. A reduction in amylose content was not achieved in any of the transformants. However, it is shown that SBDn expression can affect physical processes underlying granule assembly, in both genetic potato backgrounds, without altering the primary structure of the constituent starch polymers and the granule melting temperature. Granule size distribution of the starches obtained from transgenic Kardal plants were similar to those from untransformed controls, irrespective of the amount of SBDn accumulated. In the amf background, granule size is severely affected. In both the Kardal and amf background, apparently normal oval-shaped starch granules were composed of multiple smaller ones, as evidenced from the many “Maltese crosses” within these granules. The results are discussed in terms of different binding modes of SBD.     

Natronocalculus amylovorans gen. nov., sp. nov., and Natranaeroarchaeum aerophilus sp. nov., dominant culturable amylolytic natronoarchaea from hypersaline soda lakes in southwestern Siberia

Several pure cultures of alkaliphilic haloaloarchaea were enriched and isolated from hypersaline soda lakes in southwestern Siberia using amylopectin and fructans as substrates. Phylogenomic analysis placed the isolates into two distinct groups within the class Halobacteria. Four isolates forming group 1 were closely related to a recently described Natranaeroarchaeum sulfidigenes and the other three strains forming group 2 represent a novel genus-level phylogenetic lineage. All isolates are saccharolytic archaea growing with various starch-like alpha-glucans including soluble starch, amylopectin, dextrin, glycogen, pullulane and cyclodextrin. In addition, group 1 can use levan while group 2 – inulin (plant storage beta-fructans). Group 1 strains can also grow anaerobically with either glucose or maltose using elemental sulfur as the electron acceptor. Both groups are moderately alkaliphilic with a pH range for growth from 7.2 to 9.3 (optimum between 8.0–8.8) and low Mg-demanding extreme halophiles growing optimally at 4 M total Na⁺. The major respiratory menaquinone is MK-8:8 and the core biphytanyl lipids are dominated by archaeol (C₂₀-C₂₀) and a less abundant extended archaeol (C₂₀-C₂₅) with PG and PGP-Me as polar groups. The four isolates of group 1 are suggested to be classified into a new species as Natranaeroarchaeum aerophilus sp. nov. (type strain AArc-St1-1^T = JCM 32519^T). The three isolates of group 2 are proposed to form a new genus and species for which the name Natronocalculus amylovorans gen. nov., sp. nov. is suggested (type strain AArc-St2^T = JCM 32475^T).     

Crystal Structure of Barley Limit Dextrinase-Limit Dextrinase Inhibitor (LD-LDI) Complex Reveals Insights into Mechanism and Diversity of Cereal Type Inhibitors

Molecular details underlying regulation of starch mobilization in cereal seed endosperm remain unknown despite the paramount role of this process in plant growth. The structure of the complex between the starch debranching enzyme barley limit dextrinase (LD), hydrolyzing α-1,6-glucosidic linkages, and its endogenous inhibitor (LDI) was solved at 2.7 Å. The structure reveals an entirely new and unexpected binding mode of LDI as compared with previously solved complex structures of related cereal type family inhibitors (CTIs) bound to glycoside hydrolases but is structurally analogous to binding of dual specificity CTIs to proteases. Site-directed mutagenesis establishes that a hydrophobic cluster flanked by ionic interactions in the protein-protein interface is vital for the picomolar affinity of LDI to LD as assessed by analysis of binding by using surface plasmon resonance and also supported by LDI inhibition of the enzyme activity. A phylogenetic analysis identified four LDI-like proteins in cereals among the 45 sequences from monocot databases that could be classified as unique CTI sequences. The unprecedented binding mechanism shown here for LDI has likely evolved in cereals from a need for effective inhibition of debranching enzymes having characteristic open active site architecture. The findings give a mechanistic rationale for the potency of LD activity regulation and provide a molecular understanding of the debranching events associated with optimal starch mobilization and utilization during germination. This study unveils a hitherto not recognized structural basis for the features endowing diversity to CTIs.     

非洲紫罗兰叶片脱分化过程中核酸蛋白质和滨粉含量动态的研究

本文测定了非洲紫罗兰（Ｓａｉｎｔｐａｕｌｔａｉｏｎａｎｔｈａ）叶片脱分化过程中核酸、蛋白质和波粉的含量。结果表明：发生脱分化的叶片中的蛋白质和淀粉含量均低于对照,而ＲＮＡ含量则高于对照,ＤＮＡ含量无明显差异。叶片培养的第一天内,发生脱分化的叶片中的蛋白质含量明显下降,对照中的蛋白质含量上升。脱分化过程中,淀粉含量有一个上升、下降、再上升的变化过程,对照中淀粉含量一直上升。     

Increasing maize seed weight by enhancing the cytoplasmic ADP-glucose pyrophosphorylase activity in transgenic maize plants

ADP-glucose pyrophosphorylase (AGPase) plays a key role in regulating starch biosynthesis in cereal seeds and is likely the most important determinant of seed strength. The Escherichia coli mutant glgC gene (glgC16), which encodes a highly active and allosterically insensitive AGPase, was introduced into maize (Zea mays L.) under the control of an endosperm-specific promoter. Developing seeds from transgenic maize plants showed up to 2–4-fold higher levels of AGPase activity in the presence of 5 mM inorganic phosphate (Pi). Transgenic plants with higher cytoplasmic AGPase activity under Pi-inhibitory conditions showed increases (13–25%) in seed weight over the untransformed control. In addition, in all transgenic maize plants, the seeds were fully filled, and the seed number of transgenic plants had no significant difference compared with that of untransformed control. These results indicate that increasing cytoplasmic AGPase activity has a marked effect on sink activity and, in turn, seed weight in transgenic maize plants.     

An unusual pattern of carbohydrate utilization in Corallococcus (Myxococcus) coralloides (Myxobacterales)

The myxobacterium, Corallococcus (Myxococcus) coralloides strain Cc c127, could not utilize mono- and disaccharides, but maltotriose and the polysaccharides starch, amylose, amylopectin, and pullulan stimulated growth. Radioactive CO₂ was set free from ¹⁴C-labeled starch. When starch was degraded, small amounts of maltose and glucose accumulated in the culture supernatant. Maltotriose, however, appeared only temporarily. Outside the cells, the trisaccharide could not be split into glucose and maltose. Pullulan was hydrolyzed exclusively into a trisaccharide which during growth was immediately consumed. Hexokinase, glucose-6-phosphate dehydrogenase, 6-phosphogluconate dehydrogenase and phosphoglucomutase could readily be demonstrated in cell extracts, but fructose-1,6-diphosphate aldolase was present with low activity only. The data suggest that intracellular glucose is metabolized mainly via the pentose phosphate pathway.Prof. Dr. Gerhard Drews gratefully dedicated to his 60th birthday     

New starches from traditional Chinese medicine (TCM)--Chinese yam (Dioscorea opposita Thunb.) cultivars

The starches separated from two different Dioscorea opposita Thunb. cultivars were investigated for morphological, thermal and crystal properties. The shape of starch granules separated from different D. opposita Thunb. cultivars varied from round to oval or irregular. The surface of the starch granules appeared to be smooth without any fissures. The average particle diameter of starches from different D. opposita Thunb. cultivars was 40.3 and 38.7mum for D. 47 and D. SXY starch, respectively. The transition temperatures (T(o), T(p) and T(c)) and enthalpy of gelatinization (DeltaH(gel)) were determined using differential scanning calorimetry (DSC). The D. SXY starch showed the lower T(o) (74.2 degrees C) and the broader R (12.4). T(p) and T(c) of starch from D. 47 were higher than that of D. SXY starch. DeltaH(gel) values (11.37J/g) of D. 47 was higher than that of D. SXY starch (10.78J/g). The crystal type of starches separated from two different D. opposita cultivars was a typical C-type pattern. The degree of crystallinity of two D. opposita cultivars starches was about 45.9% and 31.5%, respectively.     

Interactive effects of phosphate deficiency, sucrose and light/dark conditions on gene expression of UDP-glucose pyrophosphorylase in Arabidopsis

The effects of inorganic phosphate (Pi) status, light/dark and sucrose on expression of UDP-glucose pyrophosphorylase (UGPase) gene (Ugp), which is involved in sucrose/ polysaccharides metabolism, were investigated using Arabidopsis wild-type (wt) plants and mutants impaired in Pi and carbohydrate status. Generally, P-deficiency resulted in increased Ugp expression and enhanced UGPase activity and protein content, as found for wt plants grown on P-deficient and complete nutrient solution, as well as for pho1 (P-deficient) mutants. Ugp was highly expressed in darkened leaves of pho1, but not wt plants; daily light exposure enhanced Ugp expression both in wt and pho mutants. The pho1 and pho2 (Pi-accumulating) mutations had little or no effect on leaf contents of glucose and fructose, regardless of light/dark conditions, whereas pho1 plants had much higher levels of sucrose and starch in the dark than pho2 and wt plants. The Ugp was up-regulated when leaves were fed with sucrose in wt plants, but the expression in pho2 background was much less sensitive to sucrose supply than in wt and pho1 plants. Expression of Ugp in pgm1 and sex1 mutants (impaired in starch/sugar content) was not dependent on starch content, and not tightly correlated with soluble sugar status. Okadaic acid (OKA) effectively blocked the P-starvation and sucrose-dependent expression of Ugp in excised leaves, whereas staurosporine (STA) had only a small effect on both processes (especially in -P leaves), suggesting that P-starvation and sucrose effects on Ugp are transmitted by pathways that may share similar components with respect to their (in) sensitivity to OKA and STA. The results of this study suggest that Ugp expression is modulated by an interaction of signals derived from P-deficiency status, sucrose content and dark/light conditions, and that light/sucrose and P-deficiency may have additive effects on Ugp expression.