Purification and characterization of sucrose synthase isozymes from etiolated rice seedlings

Huang, Der-Yi; Wang, Ai‐Yu

doi:10.1080/15216549800203612

Cited by 9 publications

(8 citation statements)

References 12 publications

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“…Mg 2+ or other cations have been frequently reported to either positively (Morell and Copeland 1985) or negatively (Tsai 1974;Huang and Wang 1998) influence the activity of SuSy in the Suc cleavage direction. To scrutinize the effect of cations on the different SuSy enzymes, the activity was determined in the presence of 200 mM Suc, 5 mM ADP, and varying concentrations of MgCl 2 (Fig.…”

Section: Phylogenetic Analysis and Expression Of Prokaryotic Susy Seqmentioning

confidence: 99%

Identification of sucrose synthase in nonphotosynthetic bacteria and characterization of the recombinant enzymes

Diricks

Bruyn

Daele

et al. 2015

Appl Microbiol Biotechnol

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Sucrose synthase (SuSy) catalyzes the reversible conversion of sucrose and a nucleoside diphosphate into fructose and nucleotide (NDP)-glucose. To date, only SuSy's from plants and cyanobacteria, both photosynthetic organisms, have been characterized. Here, four prokaryotic SuSy enzymes from the nonphotosynthetic organisms Nitrosomonas Europaea (SuSyNe), Acidithiobacillus caldus (SuSyAc), Denitrovibrio acetiphilus (SusyDa), and Melioribacter roseus (SuSyMr) were recombinantly expressed in Escherichia coli and thoroughly characterized. The purified enzymes were found to display high-temperature optima (up to 80 °C), high activities (up to 125 U/mg), and high thermostability (up to 15 min at 60 °C). Furthermore, SuSyAc, SuSyNe, and SuSyDa showed a clear preference for ADP as nucleotide, as opposed to plant SuSy's which prefer UDP. A structural and mutational analysis was performed to elucidate the difference in NDP preference between eukaryotic and prokaryotic SuSy's. Finally, the physiological relevance of this enzyme specificity is discussed in the context of metabolic pathways and genomic organization.

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Section: Phylogenetic Analysis and Expression Of Prokaryotic Susy Seqmentioning

confidence: 99%

Identification of sucrose synthase in nonphotosynthetic bacteria and characterization of the recombinant enzymes

Diricks

Bruyn

Daele

et al. 2015

Appl Microbiol Biotechnol

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“…The method is based on protocols used for the isolation of Sus from different plant species (Sebkova et al, 1995;Huang and Wang, 1998;Tanase and Yamaki, 2000;Schafer et al, 2005). Briefly, maize kernels were ground in liquid nitrogen, and proteins were extracted by incubation in extraction buffer (50 mM imidazole, pH 7.5, 1 mM EDTA, 1 mM PMSF, 1% polyvinylpyrrolidone-360, 2 mM DTT, and 10 mg/mL protease inhibitors) for 45 min at 4°C.…”

Section: Isolation Of Sus From Maize Kernels and Pollen Tubesmentioning

confidence: 99%

Sucrose Synthase Is Associated with the Cell Wall of Tobacco Pollen Tubes

et al. 2008

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Sucrose synthase (Sus; EC 2.4.1.13) is a key enzyme of sucrose metabolism in plant cells, providing carbon for respiration and for the synthesis of cell wall polymers and starch. Since Sus is important for plant cell growth, insights into its structure, localization, and features are useful for defining the relationships between nutrients, growth, and cell morphogenesis. We used the pollen tube of tobacco (Nicotiana tabacum) as a cell model to characterize the main features of Sus with regard to cell growth and cell wall synthesis. Apart from its role during sexual reproduction, the pollen tube is a typical tip-growing cell, and the proper construction of its cell wall is essential for correct shaping and direction of growth. The outer cell wall layer of pollen tubes consists of pectins, but the inner layer is composed of cellulose and callose; both polymers require metabolic precursors in the form of UDP-glucose, which is synthesized by Sus. We identified an 88-kD polypeptide in the soluble, plasma membrane and Golgi fraction of pollen tubes. The protein was also found in association with the cell wall. After purification, the protein showed an enzyme activity similar to that of maize (Zea mays) Sus. Distribution of Sus was affected by brefeldin A and depended on the nutrition status of the pollen tube, because an absence of metabolic sugars in the growth medium caused Sus to distribute differently during tube elongation. Analysis by bidimensional electrophoresis indicated that Sus exists as two isoforms, one of which is phosphorylated and more abundant in the cytoplasm and cell wall and the other of which is not phosphorylated and is specific to the plasma membrane. Results indicate that the protein has a role in the construction of the extracellular matrix and thus in the morphogenesis of pollen tubes.

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“…Attempts at producing amylopectin in vitro, however, have been unsuccessful, producing instead an animal-like glycogen product (Guan et al, 1995). In addition to the exact nature of amylopectin formation, several enzymes involved in starch biosynthesis are also poorly understood because many of these enzymes have multiple isoforms Gao et al, 1996;Huang and Wang, 1998;Sidebottom et al, 1998;Beckles et al, 2001).…”

Section: Introductionmentioning

confidence: 99%

Dissection of Maize Kernel Composition and Starch Production by Candidate Gene Association

et al. 2004

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Cereal starch production forms the basis of subsistence for much of the world's human and domesticated animal populations. Starch concentration and composition in the maize (Zea mays ssp mays) kernel are complex traits controlled by many genes. In this study, an association approach was used to evaluate six maize candidate genes involved in kernel starch biosynthesis: amylose extender1 (ae1), brittle endosperm2 (bt2), shrunken1 (sh1), sh2, sugary1, and waxy1. Major kernel composition traits, such as protein, oil, and starch concentration, were assessed as well as important starch composition quality traits, including pasting properties and amylose levels. Overall, bt2, sh1, and sh2 showed significant associations for kernel composition traits, whereas ae1 and sh2 showed significant associations for starch pasting properties. ae1 and sh1 both associated with amylose levels. Additionally, haplotype analysis of sh2 suggested this gene is involved in starch viscosity properties and amylose content. Despite starch concentration being only moderately heritable for this particular panel of diverse maize inbreds, high resolution was achieved when evaluating these starch candidate genes, and diverse alleles for breeding and further molecular analysis were identified.

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Purification and characterization of sucrose synthase isozymes from etiolated rice seedlings

Cited by 9 publications

References 12 publications

Identification of sucrose synthase in nonphotosynthetic bacteria and characterization of the recombinant enzymes

Identification of sucrose synthase in nonphotosynthetic bacteria and characterization of the recombinant enzymes

Sucrose Synthase Is Associated with the Cell Wall of Tobacco Pollen Tubes

Dissection of Maize Kernel Composition and Starch Production by Candidate Gene Association

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