Characterization of substrate and product specificity of the purified recombinant glycogen branching enzyme of Rhodothermus obamensis

Roussel, Xavier; Lancelon‐Pin, Christine; Viksø-Nielsen, Anders; Rolland‐Sabaté, Agnès; Grimaud, Florent; Potocki-Véronèse, Gabrielle; Buléon, Alain; Putaux, Jean‐Luc; d’Hulst, Christophe

doi:10.1016/j.bbagen.2012.09.022

Cited by 66 publications

(30 citation statements)

References 31 publications

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“…1D). These results are in agreement with previous investigations (Kittisuban, Lee, Suphantharika and Hamaker, 2014;Roussel et al, 2013) using R. obamensis BE with selected types of starches as substrate. Interestingly, the chains in the BEAMBE modified starches clearly showed a distributional trend characterized by an increase in short chains of DP 3-12 with higher AO substrate content (Fig.…”

Section: Chain-length Distributionsupporting

confidence: 83%

“…These results are in agreement with those obtained by Shinohara et al (2001) who suggested that R. obamensis BE has a six-fold higher activity for amylose than for amylopectin. However, another study (Roussel et al, 2013), using synthetic amylose DP 2-60 as a substrate for R.…”

Section: α-16 Glucosidic Linkagesmentioning

confidence: 99%

“…Starch modification with BE demonstrates an increase in solubility, reduced viscosity and for AM, increase degradative resistance of the product (Blennow et al, 2013). The thermostable BE from R. obamensis has drawn interest in starch modification to efficiently produce highly-branched α-glucans (Roussel et al, 2013;Shinohara et al, 2001). Both amylose and amylopectin have been tested as substrate to identify possible different mechanisms.…”

Section: Introductionmentioning

confidence: 99%

“…However, the substrate used was amylose from starch fractionation ) which had amylopectin contamination or was enzymatic synthesized glucans containing DP 2-60 (Roussel et al, 2013), none of which the chain length was long enough compared to natural amylose. Different types of natural starch differ in their amylose content but there is only limited information on the effect of amylose on BE catalysed formation of α-1,6 glucosidic linkages and cyclo-glucans (Andersson, Rydberg, Larsson, Andersson and Åman, 2002;Roussel et al, 2013;Takata et al, 1997;. In a previous study, we modified cassava starch by using combinations of AM and BE to increase the content of α-1,6 branch points.…”

Section: Introductionmentioning

confidence: 99%

“…AM has been proven to add value to starch including its modification to impose gelatin-like functionality (Hansen et al, 2008) and increased degradative resistance (Jiang, Miao, Ye, Jiang and Zhang, 2014). Branching enzyme (BE, 1,4-α-D-α-glucan:1,4-α-D-α-glucan 6-α-D-(1,4-α-D-α-glucano)-transferase, EC 2.4.1.18, glucosyl hydrolase family 13, GH13, www.CaZy.org) catalyses the formation of branch points in starch and glycogen biosynthesis by the cleavage of existing α-1,4 glucosidic linkages followed by transfer to 1,6 hydroxyl groups to form the tree-like amylopectin and glycogen structures (Roussel et al, 2013;Shinohara et al, 2001). In vitro, BE also catalyses a cyclisation reaction to form cyclo-amylose and cylo-amylopectin cluster (Kelly, Dijkhuizen and Leemhuis, 2009;.…”

Section: Introductionmentioning

confidence: 99%

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Structure of branching enzyme- and amylomaltase modified starch produced from well-defined amylose to amylopectin substrates

Sorndech¹,

Sagnelli

Meier

et al. 2016

Carbohydrate Polymers

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Highlights •Enzyme modification of starch systems with well-defined amylose:amylopectin ratios provides detailed insight into the effects of substrate branching on enzymatic chain transfer • High amylose substrates produce high branching rates, high M w, short chains and low amylolytic digestion when treated with branching enzyme alone or in combination with amylomaltase.• Not only branching density but also molar mass of the glucan product restricts dietary degradation by steric hindrance towards human pancreatic α-amylase and α-glucosidases. AbstractThermostable branching enzyme (BE, EC 2.4.1.18) from Rhodothermus obamensis in combination with amylomaltase (AM, EC 2.4.1.25) from Thermus thermophilus was used to modify starch structure exploring potentials to extensively increase the number of branch points in starch. Amylose is an important constituent in starch and the effect of amylose on enzyme catalysis was investigated using amylose-only barley starch (AO) and waxy maize starch (WX) in well-defined ratios. All products were analysed for amylopectin chain length distribution, α-1,6 glucosidic linkages content, molar mass distribution and digestibility by using rat intestinal α-glucosidases. For each enzyme treatment series, increased AO content resulted in a higher rate of α-1,6 glucosidic linkage formation but as an effect of the very low initial branching of the AO, the final content of α-1,6 glucosidic linkages was slightly lower as compared to the high amylopectin substrates. However, an increase specifically in short chains was produced at high AO levels. The molar mass distribution for the enzyme treated samples was lower as compared with substrate WX and AO, indicating the presence of hydrolytic activity as well as cyclisation of the substrate. For all samples, increased amylose substrate showed decreased α-and β-amylolysis. Surprisingly, hydrolysis with rat intestinal α-glucosidases was higher with increasing α-1,6 glucosidic linkage content and decreasing M w indicating that steric hindrance towards the α-glucosidases was directed by the molar mass rather that the branching density of the glucan per se. Our data demonstrate that a higher amylose content in the substrate starch efficiently produces α-1,6 glucosidic linkages and that the present of amylose generates a higher M w and more resistant product than amylopectin. The combination of BEAMBE provided somewhat more resistant α-glucan products as compared to BE alone.

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Section: Chain-length Distributionsupporting

confidence: 83%

Section: α-16 Glucosidic Linkagesmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Structure of branching enzyme- and amylomaltase modified starch produced from well-defined amylose to amylopectin substrates

Sorndech¹,

Sagnelli

Meier

et al. 2016

Carbohydrate Polymers

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Polysaccharide Biocatalysis: From Synthesizing Carbohydrate Standards to Establishing Characterization Methods

Ćirić

Petrović

Loos

2014

Macro Chemistry & Physics

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Starch, glycogen, and cellulose are all around us. They are eaten and used on a daily basis but they are not understood completely. Even though these carbohydrates are simple, concerning their repeating unit, they are hard to characterize. In order to try to understand as much as possible about their structure and the relationship between their molecular structure and physical properties, it is very practical to create such polysaccharides, for instance enzymatically, characterize them, and use them as standards for the characterization of natural ones. Therefore, the main objective of this Trend article is to outline different enzymatic routes to such carbohydrates, possibilities for their characterization, and the characterization of natural ones.

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A glycogen branching enzyme from Thermomonospora curvata: Characterization and its action on maize starch

et al. 2015

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A gene (TcGBE) encoding a glycogen branching enzyme from Thermomonospora curvata was expressed in Escherichia coli and purified, and biochemically analyzed. The purified enzyme was approximately 84 kDa, and exhibited a maximum activity at 55°C and pH 8.5. The enzyme showed the highest specificity to amylose with a specific activity of 90.28 U/mg. The recombinant enzyme could act on maize starch, and starch treated with TcGBE was observed to have a lower amylose content and molecular weight. The number of short chains of amylopectin in TcGBEmodified starch with a degree of polymerization (DP) less than 13 increased from 20.19 to 32.59%, and was accompanied by a reduction in longer chains (DP > 25) from 25.58 to 10.03%. In addition, the medium length chains of amylopectin (13 < DP <25) also showed a slight increase from 54.23 to 58.38%. The 1 H NMR spectra revealed an increased ratio of a-1,6-to a-1, 4-glucosidic linkages from 4.1 to 10.3% following treatment with TcGBE for 10 h. Taken together, these results suggested that TcGBE could be used to generate a modified maize starch with a low amylose content and more branched amylopectin of low molecular weight but richer in short and intermediate chains.

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Characterization of substrate and product specificity of the purified recombinant glycogen branching enzyme of Rhodothermus obamensis

Cited by 66 publications

References 31 publications

Structure of branching enzyme- and amylomaltase modified starch produced from well-defined amylose to amylopectin substrates

Structure of branching enzyme- and amylomaltase modified starch produced from well-defined amylose to amylopectin substrates

Polysaccharide Biocatalysis: From Synthesizing Carbohydrate Standards to Establishing Characterization Methods

A glycogen branching enzyme from Thermomonospora curvata: Characterization and its action on maize starch

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