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To investigate the functional properties of 10 α-glucan branching enzymes (BEs) from various sources, we determined the chain-length distribution of BE enzymatic products and their phosphorylase-limit dextrins (Φ-LD). All BEs could be classified into either of the three rice BE isozymes: OsBEI, OsBEIIa, or OsBEIIb. Escherichia coli BE (EcoBE) had the same enzymatic properties as OsBEI, while Synechococcus elongatus BE (ScoBE) and Chlorella kessleri BE (ChlBE) had BEIIb-type properties. Human BE (HosBE), yeast BE (SacBE), and two Porphyridium purpureum BEs (PopBE1 and PopBE2) exhibited the OsBEIIa-type properties. Analysis of chain-length profile of Φ-LD of the BE reaction products revealed that EcoBE, ScoBE, PopBE1, and PopBE2 preferred A-chains as acceptors, while OsBEIIb used B-chains more frequently than A-chains. Both EcoBE and ScoBE specifically formed the branch linkages at the third glucose residue from the reducing end of the acceptor chain. The present results provide evidence for the first time that great variation exists as to the preference of BEs for their acceptor chain, either A-chain or B-chain. In addition, EcoBE and ScoBE recognize the location of branching points in their acceptor chain during their branching reaction. Nevertheless, no correlation exists between the primary structure of BE proteins and their enzymatic characteristics. [ABSTRACT FROM AUTHOR]

Up to now, no information on the structural characteristics of glucans involved in the initiation of starch biosynthesis in cereal endosperm has been available. Since amylopectin is a major constituent of starch and largely affects the starch physicochemical properties, we attempted to characterize the fine structures of branched glucans. We isolated small glucan granules with a diameter of approximately 40–80 nm from rice endosperm at the early developmental stages and analyzed the chain-length distribution of the glucans and their phosphorylase-limit dextrins. Compared with normal amylopectin, the proportion of long chains of these glucans with degree of polymerization (DP) ≥ 35 was significantly lower and these long chains did not form a peak. These structural features were commonly found in small glucan granules in three rice cultivars. Based on the present results, we propose that these immature glucans located in small granules in young endosperm are involved in the initiation process of amylopectin biosynthesis in rice endosperm. • Isolation and characterization of immature glucans in very young rice endosperm. • The glucans having structurally an immature cluster compared with amylopectin. • Possible role of immature glucans in the initial starch synthesis in rice endosperm. • New insights into the specific initial stage of starch synthesis in cereal endosperm. [ABSTRACT FROM AUTHOR]

Abstract : Through integrated molecular and biochemical investigations and by using a common mutant line in molecular mapping, we have shown that the SSIIa gene, previously reported to be responsible for the starch granule gelatinization temperature (GT) differences between indica and japonica rice varieties, might also control the GT variations among certain indica varieties. This effect is mediated through differences in the amounts of SSIIa protein bound to starch granules leading to differences in the structure of amylopectin molecules synthesized. For the first time it was shown, that the amylopectin type accounted only for the GT differences between varieties with different SSIIa alleles, but not between varieties carrying a common SSIIa allele. In the latter case, another gene, alk2(t), with a genetic distance of 3.93cM from the Wx gene, was identified as being responsible for the GT variations. Based on the thermal properties and amylopectin chain length profile characteristics, it is postulated that the SSIIa gene has at least three different alleles, one in japonica rice and two in indica rice varieties, whereas alk2(t) has at least two alleles for either low or high GT. Thus, the rich diversity of the GT character in rice very probably results from various combinations of these alleles. [Copyright &y& Elsevier]

Starch debranching enzyme (DBE)-deficient, sugary-1 mutations of rice caused the biosynthesis of more highly branched α-glucan in the form of phytoglycogen and sugary -amylopectin rather than wild-type amylopectin. Phytoglycogen and sugary -amylopectin are located in the inner and outer regions of the rice endosperm, respectively. Detailed analyses of these glucans from a variety of allelic mutants with different severity of sugary phenotype revealed that sugary -amylopectins consisted of more short chains of DP<12 but fewer chains of DP13–24 than did wild-type amylopectin, and the alterations reflected on lower molecular weights, smaller gyration radii, and denser molecules. These changes resulted in lower onset gelatinization temperature, lower peak viscosity, lesser degree of crystallinity, and smaller and cracked granules with rough surface of sugary starch granules as compared with that of the wild-type. These results indicate that changes in the fine structure of amylopectin induced by lesion of DBE greatly influence the morphological and physicochemical properties of starch. Phytoglycogens from sugary-1 mutants were composed of multiple components of smaller molecular weights and gyration radii than sugary -amylopectin and amylopectin. The highly branched phytoglycogens were more tightly packed molecules exhibiting greater dispersed densities than sugary -amylopectin and amylopectin. These results suggest that the levels of DBE in rice endosperm control the biosynthetic pathway of branched α-glucans and produce amylopectin, sugary -amylopectin and phytoglycogen. [Copyright &y& Elsevier]