Determination of alginate composition by a simple enzymatic assay

�stgaard, Kjetill

doi:10.1007/bf00049064

Cited by 25 publications

(8 citation statements)

References 20 publications

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“…Alginate lyases from bacterial sources have been used in analyzing alginate structure, protoplasting of seaweed, studying Fucus cell wall development, and producing polyM, or polyG blocks [16,17,18,19,20]. Recently alginate oligomers with the specific molecular weight have been shown to have specific bio-activities, such as stimulating the secretion of cytotoxic cytokines from human macrophage [21] or promoting the growth of bifidobacteria [22].…”

Section: Discussionmentioning

confidence: 99%

Cultivable Alginate Lyase-Excreting Bacteria Associated with the Arctic Brown Alga Laminaria

et al. 2012

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Although some alginate lyases have been isolated from marine bacteria, alginate lyases-excreting bacteria from the Arctic alga have not yet been investigated. Here, the diversity of the bacteria associated with the brown alga Laminaria from the Arctic Ocean was investigated for the first time. Sixty five strains belonging to nine genera were recovered from six Laminaria samples, in which Psychrobacter (33/65), Psychromonas (10/65) and Polaribacter (8/65) were the predominant groups. Moreover, 21 alginate lyase-excreting strains were further screened from these Laminaria-associated bacteria. These alginate lyase-excreting strains belong to five genera. Psychromonas (8/21), Psedoalteromonas (6/21) and Polaribacter (4/21) are the predominant genera, and Psychrobacter, Winogradskyella, Psychromonas and Polaribacter were first found to produce alginate lyases. The optimal temperatures for the growth and algiante lyase production of many strains were as low as 10–20 °C, indicating that they are psychrophilic bacteria. The alginate lyases produced by 11 strains showed the highest activity at 20–30 °C, indicating that these enzymes are cold-adapted enzymes. Some strians showed high levels of extracellular alginate lyase activity around 200 U/mL. These results suggest that these algiante lyase-excreting bacteria from the Arctic alga are good materials for studying bacterial cold-adapted alginate lyases.

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Section: Discussionmentioning

confidence: 99%

Cultivable Alginate Lyase-Excreting Bacteria Associated with the Arctic Brown Alga Laminaria

et al. 2012

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“…Aasen et al [1] have developed methods for the large scale purification of the Kleb-sieUa alginate lyase for such applications. The same group proposed the combined use of the Klebsiella guluronate lyase and a polymannuronate lyase from Haliotis tuberculata for a spectrophotometric assay for alginates [87]. The application of the two enzymes on their own and in combination was claimed to provide an analysis of the block structure of alginates.…”

Section: Lyases and Their Products As Biotechnological Tools?mentioning

confidence: 99%

Polysaccharide lyases

Sutherland

1995

FEMS Microbiology Reviews

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Polysaccharide lyases are the products of various microorganisms, bacteriophage and some eukaryotes. All such enzymes cleave a hexose-1,4-alpha- or beta-uronic acid sequence by beta-elimination. They are in some examples, the only known type of enzymes degrading their polyanionic substrates. Although only a small number of these enzymes have been exhaustively studied, the pectin lyases of bacterial origin have proved to be of interesting crystal structure containing a parallel beta-helix domain. Alginate and heparin lyases may yield products with biotechnological potential.

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“…Alginate was quantified enzymatic essentially as described earlier in [12]. The cultures in the replicate used for enzymatic alginate measurements were supplemented with 0.05 mL/L alkalase 2.4 L and neutrase 0.5 L (Novo Nordisk) during cultivation to avoid extracellular degradation of alginate.…”

Section: Cultivation Of the Mutant Strains Enzymatic Alginate Quantimentioning

confidence: 99%

Rapid reagentless quantification of alginate biosynthesis in Pseudomonas fluorescens bacteria mutants using FT-IR spectroscopy coupled to multivariate partial least squares regression

et al. 2012

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Alginate is an important medical and commercial product and currently is isolated from seaweeds. Certain microorganisms also produce alginate and these polymers have the potential to replace seaweed alginates in some applications, mainly because such production will allow much better and more reproducible control of critical qualitative polymer properties. The research conducted here presents the development of a new approach to this problem by analysing a transposon insertion mutant library constructed in an alginate-producing derivative of the Pseudomonas fluorescens strain SBW25. The procedure is based on the non-destructive and reagent-free method of Fourier transform infrared (FT-IR) spectroscopy which is used to generate a complex biochemical infrared fingerprint of the medium after bacterial growth. First, we investigate the potential differences caused by the growth media fructose and glycerol on the bacterial phenotype and alginate synthesis in 193 selected P. fluorescens mutants and show that clear phenotypic differences are observed in the infrared fingerprints. In order to quantify the level of the alginate we also report the construction and interpretation of multivariate partial least squares regression models which were able to quantify alginate levels successfully with typical normalized root-mean-square error in predictions of only approximately 14%. We have demonstrated that this high-throughput approach can be implemented in alginate screens and we believe that this FT-IR spectroscopic methodology, when combined with the most appropriate chemometrics, could easily be modified for the quantification of other valuable microbial products and play a valuable screening role for synthetic biology.

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Determination of alginate composition by a simple enzymatic assay

Cited by 25 publications

References 20 publications

Cultivable Alginate Lyase-Excreting Bacteria Associated with the Arctic Brown Alga Laminaria

Cultivable Alginate Lyase-Excreting Bacteria Associated with the Arctic Brown Alga Laminaria

Polysaccharide lyases

Rapid reagentless quantification of alginate biosynthesis in Pseudomonas fluorescens bacteria mutants using FT-IR spectroscopy coupled to multivariate partial least squares regression

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