A rapid, sensitive, simple plate assay for detection of microbial alginate lyase activity

Sawant, Shailesh S.; Salunke, Bipinchandra K.; Kim, Beom Soo

doi:10.1016/j.enzmictec.2015.05.003

Cited by 44 publications

(22 citation statements)

References 40 publications

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“…Detection of alginate lyase activity in cultures grown on solid media was performed, with modifications59. Strains were grown on PIA medium for 72 h. Biomass was suspended in saline and cells were pelleted by centrifugation.…”

Section: Methodsmentioning

confidence: 99%

Biological function of a polysaccharide degrading enzyme in the periplasm

Wang

Moradali

Goudarztalejerdi

et al. 2016

Sci Rep

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Carbohydrate polymers are industrially and medically important. For instance, a polysaccharide, alginate (from seaweed), is widely used in food, textile and pharmaceutical industries. Certain bacteria also produce alginate through membrane spanning multi-protein complexes. Using Pseudomonas aeruginosa as a model organism, we investigated the biological function of an alginate degrading enzyme, AlgL, in alginate production and biofilm formation. We showed that AlgL negatively impacts alginate production through its enzymatic activity. We also demonstrated that deletion of AlgL does not interfere with polymer length control, epimerization degree or stability of the biosynthesis complex, arguing that AlgL is a free periplasmic protein dispensable for alginate production. This was further supported by our protein-stability and interaction experiments. Interestingly, over-production of AlgL interfered with polymer length control, suggesting that AlgL could be loosely associated with the biosynthesis complex. In addition, chromosomal expression of algL enhanced alginate O-acetylation; both attachment and dispersal stages of the bacterial biofilm lifecycle were sensitive to the level of O-acetylation. Since this modification also protects the pathogen against host defences and enhances other virulence factors, chromosomal expression of algL could be important for the pathogenicity of this organism. Overall, this work improves our understanding of bacterial alginate production and provides new knowledge for alginate production and disease control.

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Section: Methodsmentioning

confidence: 99%

Biological function of a polysaccharide degrading enzyme in the periplasm

Wang

Moradali

Goudarztalejerdi

et al. 2016

Sci Rep

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show abstract

“…The new plate was incubated at 30 °C to form visible colonies, while the original plates were stained with Gram’s iodine solution (Sigma, St. Louis, USA, catalogue HT902). The clear zone around the strain colony indicated that the strain secreted alginate lyase into the medium (Sawant et al 2015). The number of strains with clear zone was recorded and these strains were selected for further analysis.…”

Section: Methodsmentioning

confidence: 99%

“…The existing methods are still low efficient for revealing alginate lyase-excreting microbes at large scale. Recently, Sawant et al (2015) reported that Gram’s iodine was an excellent chromogenic agent for revealing alginate lyase-producing colonies on agar plates. According Sawant’s result, Gram’s iodine could form distinct clear zones around the alginate degrading microbial colonies within 2–3 min.…”

Section: Introductionmentioning

confidence: 99%

Screening of alginate lyase-excreting microorganisms from the surface of brown algae

et al. 2017

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Alginate lyase is a biocatalyst that degrades alginate to produce oligosaccharides, which have many bioactive functions and could be used as renewable biofuels. Here we report a simple and sensitive plate assay for screening alginate lyase-excreting microorganisms from brown algae. Brown algae Laminaria japonica, Sargassum horneri and Sargassum siliquatrum were cultured in sterile water. Bacteria growing on the surface of seaweeds were identified and their capacity of excreting alginate lyase was analyzed. A total of 196 strains were recovered from the three different algae samples and 12 different bacterial strains were identified capable of excreting alginate lyases. Sequence analysis of the 16S rRNA gene revealed that these alginate lyase-excreting strains belong to eight genera: Paenibacillus (4/12), Bacillus (2/12), Leclercia (1/12), Isoptericola (1/12), Planomicrobium (1/12), Pseudomonas (1/12), Lysinibacillus (1/12) and Sphingomonas (1/12). Further analysis showed that the LJ-3 strain (Bacillus halosaccharovorans) had the highest enzyme activity. To our best knowledge, this is the first report regarding alginate lyase-excreting strains in Paenibacillus, Planomicrobium and Leclercia. We believe that our method used in this study is relatively easy and reliable for large-scale screening of alginate lyase-excreting microorganisms.Electronic supplementary materialThe online version of this article (doi:10.1186/s13568-017-0361-x) contains supplementary material, which is available to authorized users.

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“…Alginate lyase plate assay. Detection of alginate lyase activity in cultures grown on solid media was performed, with modifications 59 . Strains were grown on PIA medium for 72 h. Biomass was suspended in saline and cells were pelleted by centrifugation.…”

mentioning

confidence: 99%

Biological function of a polysaccharide degrading enzyme in the periplasm

Wang¹,

Moradali²,

Goudarztalejerdi³

et al. 2020

Preprint

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© The Author(s) 2016. Carbohydrate polymers are industrially and medically important. For instance, a polysaccharide, alginate (from seaweed), is widely used in food, textile and pharmaceutical industries. Certain bacteria also produce alginate through membrane spanning multi-protein complexes. Using Pseudomonas aeruginosa as a model organism, we investigated the biological function of an alginate degrading enzyme, AlgL, in alginate production and biofilm formation. We showed that AlgL negatively impacts alginate production through its enzymatic activity. We also demonstrated that deletion of AlgL does not interfere with polymer length control, epimerization degree or stability of the biosynthesis complex, arguing that AlgL is a free periplasmic protein dispensable for alginate production. This was further supported by our protein-stability and interaction experiments. Interestingly, over-production of AlgL interfered with polymer length control, suggesting that AlgL could be loosely associated with the biosynthesis complex. In addition, chromosomal expression of algL enhanced alginate O-acetylation; both attachment and dispersal stages of the bacterial biofilm lifecycle were sensitive to the level of O-acetylation. Since this modification also protects the pathogen against host defences and enhances other virulence factors, chromosomal expression of algL could be important for the pathogenicity of this organism. Overall, this work improves our understanding of bacterial alginate production and provides new knowledge for alginate production and disease control.

show abstract

A rapid, sensitive, simple plate assay for detection of microbial alginate lyase activity

Cited by 44 publications

References 40 publications

Biological function of a polysaccharide degrading enzyme in the periplasm

Biological function of a polysaccharide degrading enzyme in the periplasm

Screening of alginate lyase-excreting microorganisms from the surface of brown algae

Biological function of a polysaccharide degrading enzyme in the periplasm

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