Ultra-high performance liquid chromatography-size exclusion chromatography (UPLC-SEC) as an efficient tool for the rapid and highly informative characterisation of biopolymers

Pérez-Moral, Natalia; Plankeele, Jean-Michel; Domoney, Claire; Warren, Frederick J.

doi:10.1016/j.carbpol.2018.05.049

Cited by 17 publications

(17 citation statements)

References 28 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…As expected from the severe granule phenotype, debranched starch from the 237/4-44 mutant had a radically altered distribution of chain lengths relative to wild type. Information on the distribution of chain lengths was obtained from calibrated HPLC-SEC outputs (Perez-Moral et al, 2018;Wu et al, 2014), which provide information about weight distributions of chains as a function of hydrodynamic volume. This approach revealed three major peaks of chains in starch from control plants (protoplastderived wild-type plants; Figure 5).…”

Section: Mutations In Sbe2 or Both Sbe Genes Results In Diverse Starchmentioning

confidence: 99%

“…This approach revealed three major peaks of chains in starch from control plants (protoplastderived wild-type plants; Figure 5). The peak at dp 20 contains amylopectin chains that lie within a single crystalline lamella in the granule matrix, the peak at dp 50 contains long amylopectin chains that span crystalline lamellae, and the peak at dp 2500 contains amylose chains (Perez-Moral et al, 2018). In the 237/4-44 mutant, the short chains were strongly depleted, and the abundance and length of long amylopectin chains were increased ( Figure 5a).…”

Section: Mutations In Sbe2 or Both Sbe Genes Results In Diverse Starchmentioning

confidence: 99%

“…Chain‐length distributions were analysed by HPLC‐SEC following solubilization and enzymatic debranching of purified starch, according to Perez‐Moral et al . (). 1 H‐NMR analysis of branch‐point frequency was performed according to Tizzotti et al .…”

Section: Methodsmentioning

confidence: 97%

See 2 more Smart Citations

Cas9‐mediated mutagenesis of potato starch‐branching enzymes generates a range of tuber starch phenotypes

Tuncel

Corbin

Ahn‐Jarvis

et al. 2019

Plant Biotechnology Journal

Self Cite

116

View full text Add to dashboard Cite

Summary We investigated whether Cas9‐mediated mutagenesis of starch‐branching enzymes (SBEs) in tetraploid potatoes could generate tuber starches with a range of distinct properties. Constructs containing the Cas9 gene and sgRNAs targeting SBE1, SBE2 or both genes were introduced by Agrobacterium‐mediated transformation or by PEG‐mediated delivery into protoplasts. Outcomes included lines with mutations in all or only some of the homoeoalleles of SBE genes and lines in which homoeoalleles carried several different mutations. DNA delivery into protoplasts resulted in mutants with no detectable Cas9 gene, suggesting the absence of foreign DNA. Selected mutants with starch granule abnormalities had reductions in tuber SBE1 and/or SBE2 protein that were broadly in line with expectations from genotype analysis. Strong reduction in both SBE isoforms created an extreme starch phenotype, as reported previously for low‐SBE potato tubers. HPLC‐SEC and 1H NMR revealed a decrease in short amylopectin chains, an increase in long chains and a large reduction in branching frequency relative to wild‐type starch. Mutants with strong reductions in SBE2 protein alone had near‐normal amylopectin chain‐length distributions and only small reductions in branching frequency. However, starch granule initiation was enormously increased: cells contained many granules of <4 μm and granules with multiple hila. Thus, large reductions in both SBEs reduce amylopectin branching during granule growth, whereas reduction in SBE2 alone primarily affects numbers of starch granule initiations. Our results demonstrate that Cas9‐mediated mutagenesis of SBE genes has the potential to generate new, potentially valuable starch properties without integration of foreign DNA into the genome.

show abstract

Section: Mutations In Sbe2 or Both Sbe Genes Results In Diverse Starchmentioning

confidence: 99%

Section: Mutations In Sbe2 or Both Sbe Genes Results In Diverse Starchmentioning

confidence: 99%

See 1 more Smart Citation

Cas9‐mediated mutagenesis of potato starch‐branching enzymes generates a range of tuber starch phenotypes

Tuncel

Corbin

Ahn‐Jarvis

et al. 2019

Plant Biotechnology Journal

Self Cite

116

View full text Add to dashboard Cite

show abstract

“…Rheological measurements are also informative, since the apparent viscosity allows estimation of the amount of dissolved macromolecules/polymers (e.g., soluble polysaccharides and starch). Size-exclusion chromatography (SEC) complements this data by measuring their size and molecular structure [88,89]. Furthermore, viscoelasticity measurements provide relative magnitudes of the "solid-like" and "liquid-like" behavior of the sample.…”

Section: Overall Appraisal Of the Food Structurementioning

confidence: 99%

Plant Cell Walls: Impact on Nutrient Bioaccessibility and Digestibility

Holland

Ryden

Edwards

et al. 2020

Foods

104

View full text Add to dashboard Cite

Cell walls are important structural components of plants, affecting both the bioaccessibility and subsequent digestibility of the nutrients that plant-based foods contain. These supramolecular structures are composed of complex heterogeneous networks primarily consisting of cellulose, and hemicellulosic and pectic polysaccharides. The composition and organization of these different polysaccharides vary depending on the type of plant tissue, imparting them with specific physicochemical properties. These properties dictate how the cell walls behave in the human gastrointestinal tract, and how amenable they are to digestion, thereby modulating nutrient release from the plant tissue. This short narrative review presents an overview of our current knowledge on cell walls and how they impact nutrient bioaccessibility and digestibility. Some of the most relevant methods currently used to characterize the food matrix and the cell walls are also described.

show abstract

“…5). There is relatively little intraspecies variation in amylose chain length distributions, but large interspecies variation in the length and abundance of AM1 and AM2 chains (Wang et al ., 2014; Perez‐Moral et al ., 2018).…”

Section: Amylose Structurementioning

confidence: 99%

Amylose in starch: towards an understanding of biosynthesis, structure and function

Seung

2020

New Phytologist

147

View full text Add to dashboard Cite

Summary Starch granules are composed of two distinct glucose polymers – amylose and amylopectin. Amylose constitutes 5–35% of most natural starches and has a major influence over starch properties in foods. Its synthesis and storage occurs within the semicrystalline amylopectin matrix of starch granules, this poses a great challenge for biochemical and structural analyses. However, the last two decades have seen vast progress in understanding amylose synthesis, including new insights into the action of GRANULE BOUND STARCH SYNTHASE (GBSS), the major glucosyltransferase that synthesises amylose, and the discovery of PROTEIN TARGETING TO STARCH1 (PTST1) that targets GBSS to starch granules. Advances in analytical techniques have resolved the fine structure of amylose, raising new questions on how structure is determined during biosynthesis. Furthermore, the discovery of wild plants that do not produce amylose revives a long‐standing question of why starch granules contain amylose, rather than amylopectin alone. Overall, these findings contribute towards a full understanding of amylose biosynthesis, structure and function that will be essential for future approaches to improve starch quality in crops.

show abstract

Ultra-high performance liquid chromatography-size exclusion chromatography (UPLC-SEC) as an efficient tool for the rapid and highly informative characterisation of biopolymers

Abstract: Graphical abstract

Cited by 17 publications

References 28 publications

Cas9‐mediated mutagenesis of potato starch‐branching enzymes generates a range of tuber starch phenotypes

Cas9‐mediated mutagenesis of potato starch‐branching enzymes generates a range of tuber starch phenotypes

Plant Cell Walls: Impact on Nutrient Bioaccessibility and Digestibility

Amylose in starch: towards an understanding of biosynthesis, structure and function

Contact Info

Product

Resources

About