Acid hydrolysis and high-performance liquid chromatography of xanthan

Tait, M. I.; Sutherland, Ian W.; Clarke-Sturman, A.J.

doi:10.1016/0144-8617(90)90080-c

Cited by 16 publications

(3 citation statements)

References 9 publications

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“…Uronic acids, for example, are prone to degradation and can only be recovered partially ( 79 ). In contrast to this, dimers of uronic acids and hexoses are fairly stable, resulting in a reduced release and thereby reduced detection of attached hexose ( 78 ).…”

Section: Resultsmentioning

confidence: 99%

Tailor-made exopolysaccharides—CRISPR-Cas9 mediated genome editing in Paenibacillus polymyxa

et al. 2017

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Application of state-of-the-art genome editing tools like CRISPR-Cas9 drastically increase the number of undomesticated micro-organisms amenable to highly efficient and rapid genetic engineering. Adaptation of these tools to new bacterial families can open up entirely new possibilities for these organisms to accelerate as biotechnologically relevant microbial factories, also making new products economically competitive. Here, we report the implementation of a CRISPR-Cas9 based vector system in Paenibacillus polymyxa, enabling fast and reliable genome editing in this host. Homology directed repair allows for highly efficient deletions of single genes and large regions as well as insertions. We used the system to investigate the yet undescribed biosynthesis machinery for exopolysaccharide (EPS) production in P. polymyxa DSM 365, enabling assignment of putative roles to several genes involved in EPS biosynthesis. Using this simple gene deletion strategy, we generated EPS variants that differ from the wild-type polymer not only in terms of monomer composition, but also in terms of their rheological behavior. The developed CRISPR-Cas9 mediated engineering approach will significantly contribute to the understanding and utilization of socially and economically relevant Paenibacillus species and extend the polymer portfolio.

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Section: Resultsmentioning

confidence: 99%

Tailor-made exopolysaccharides—CRISPR-Cas9 mediated genome editing in Paenibacillus polymyxa

et al. 2017

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“…Glucose and galactose as neutral sugars along with glucuronic acid were found in the same percentages (10 to 12%), while mannose, galacturonic acid, and pyruvated mannose were present in lower concentrations, 4.4, 6.3, and 5%, respectively ( Table 3). The presence of pyruvate linked to the mannose was also determined by nuclear magnetic resonance and by comparison with xanthan (38). No mannuronic acid or guluronic acid was detected.…”

Section: Resultsmentioning

confidence: 99%

Description of a new polymer-secreting bacterium from a deep-sea hydrothermal vent, Alteromonas macleodii subsp. fijiensis, and preliminary characterization of the polymer

Raguénès

Pignet

Gauthier

et al. 1996

Appl Environ Microbiol

140

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A deep-sea, aerobic, mesophilic, heterotrophic bacterium was isolated from fluid collected near an active hydrothermal vent. On the basis of phenotypic and phylogenetic analyses and DNA-DNA relatedness, strain ST716 could be assigned to the species Alteromonas macleodii as a new subspecies. This bacterium secreted an unusual high-molecular-weight polysaccharide in the presence of glucose in batch cultures. The viscosity of this exopolysaccharide is of the same order of magnitude as that of xanthan, another bacterial polysaccharide of industrial interest. This polysaccharide, produced during the stationary phase, contained glucose, mannose, pyruvated mannose, and galactose along with galacturonic acid and glucuronic acid.

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“…Prolonging the time or harshening the conditions of hydrolysis would lead to higher degradation of uronic acids. In their work on the degradation kinetics of uronic acids during hydrolysis Tait, Sutherland, and Clarke-Sturman (1990) recommended that uronic acids can also be quantified in the form of their aldobiouronic acid (dimer of uronic acid and a hexose). This provides a significant advantage of milder hydrolysis conditions.…”

Section: Validation Of the Screening Procedures With Defined Polymer Smentioning

confidence: 99%

High throughput exopolysaccharide screening platform: From strain cultivation to monosaccharide composition and carbohydrate fingerprinting in one day

Rühmann

Schmid

Sieber

2015

Carbohydrate Polymers

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Microbial exopolysaccharides (EPS) are multifunctional biogenic polymers, which exist in highly diverse chemical structures. To facilitate a fast determination of the carbohydrate composition of novel isolated strains or modified EPS variants a fast screening and analytical method is required. The platform as realized and described in this article is based on the fast carbohydrate analysis via liquid chromatography coupled with ultra violet and electrospray ionization ion trap detection in 96-well format to detect different sugars, sugar derivatives and substituents such as pyruvate. Monosaccharide analysis from hydrolyzed polysaccharides was validated successfully by 16 commercially available polymers with known structure. The method is sensitive enough to distinguish various types of sphingans which solely differ in small alterations in the monomer composition. Even a quantitative detection of single monomers as present in complex plant polysaccharides like karaya gum, with the lowest recovery, was in accordance with literature. Furthermore, 94 bacterial strains for the validation of the screening platform were completely analyzed and 41 EPS producing strains were efficiently identified. Using the method a carbohydrate-fingerprint of the strains was obtained even allowing a very fast differentiation between strains belonging to the same species. This method can become a valuable tool not only in the fast analysis of strain isolates but also in the targeted screening for polysaccharides containing special rare sugars as well in the screening of strain libraries from genetic engineering for altered structures.

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Acid hydrolysis and high-performance liquid chromatography of xanthan

Cited by 16 publications

References 9 publications

Tailor-made exopolysaccharides—CRISPR-Cas9 mediated genome editing in Paenibacillus polymyxa

Tailor-made exopolysaccharides—CRISPR-Cas9 mediated genome editing in Paenibacillus polymyxa

Description of a new polymer-secreting bacterium from a deep-sea hydrothermal vent, Alteromonas macleodii subsp. fijiensis, and preliminary characterization of the polymer

High throughput exopolysaccharide screening platform: From strain cultivation to monosaccharide composition and carbohydrate fingerprinting in one day

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