Purification and characterization of a new alginate lyase from a marine bacterium Vibrio sp.

Wang, Ya; EnWen, Guo; Yu, Wengong; Han, Feng

doi:10.1007/s10529-012-1134-x

Cited by 42 publications

(32 citation statements)

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“…QY105, Bacillus sp. ATB-1015 strain, and Pseudomonas aeruginosa were in the range (37-41 kDa) [15,22,25] similarly to that reported by the investigated alginate lyase obtained from Pseudomonas stutzeri MSEA04 (40 kDa). …”

Section: Discussionsupporting

confidence: 83%

Purification and characterization of alginate lyase from locally isolated marinePseudomonas stutzeriMSEA04

Beltagy

El-Borai

Lewiz

et al. 2016

Acta Biologica Hungarica

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An alginate lyase with high specific enzyme activity was purified from Pseudomonas stutzeri MSEA04, isolated from marine brown algae. The alginate lyase was purified by precipitation with ammonium sulphate, acetone and ethanol individually. 70% ethanol fraction showed maximum specific activity (133.3 U/mg). This fraction was re-purified by anion exchange chromatography DEAE-Cellulose A-52. The loaded protein was separated into 3 peaks. The second protein peak was the major one which contained 48.2% of the total protein recovered and 79.4% of the total recovered activity. The collected fractions of this peak were subjected to further purification by re-chromatography on Sephadex G-100. Alginate lyase activity was fractionated in the Sephadex column into one major peak, and the specific activity of this fraction reached 116 U/mg. The optimal substrate concentration, pH and temperature for alginate lyase activity were 8 mg/ml, pH 7.5 and 37 °C, respectively. While, K m and V max values were 1.07 mg alginate/ ml and 128.2 U/mg protein, respectively. The enzyme was partially stable below 50 °C, and the activity of the enzyme was strongly enhanced by K + , and strongly inhibited by Ba +2 , Cd +2 , Fe +2 and Zn +2 . The purified enzyme yielded a single band on SDS-PAGE with molecular weight (40.0 kDa).

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Section: Discussionsupporting

confidence: 83%

Purification and characterization of alginate lyase from locally isolated marinePseudomonas stutzeriMSEA04

Beltagy

El-Borai

Lewiz

et al. 2016

Acta Biologica Hungarica

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“…The culmination of these steps results in the ␤-elimination of the glycosidic bond (3,18,20,21), which generates an exposed L-guluronate or a D-mannuronate and a nonreducing end. The nonreducing end contains a 4-deoxy-L-erythro-hex-4-enepyranosyluronate residue that is signified with ⌬ to emphasize its structural difference from both L-guluronate and D-mannuronate (22).Multiple studies have investigated alginate lyases from diverse organisms (9,11,(23)(24)(25)(26)(27)(28)(29). In this study, we investigated the alginate lyases from Vibrio splendidus 12B01 (30).…”

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confidence: 99%

“…Multiple studies have investigated alginate lyases from diverse organisms (9,11,(23)(24)(25)(26)(27)(28)(29). In this study, we investigated the alginate lyases from Vibrio splendidus 12B01 (30).…”

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confidence: 99%

Alginate Lyases from Alginate-Degrading Vibrio splendidus 12B01 Are Endolytic

Badur

Jagtap

Yalamanchili

et al. 2015

Appl Environ Microbiol

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bAlginate lyases are enzymes that degrade alginate through ␤-elimination of the glycosidic bond into smaller oligomers. We investigated the alginate lyases from Vibrio splendidus 12B01, a marine bacterioplankton species that can grow on alginate as its sole carbon source. We identified, purified, and characterized four polysaccharide lyase family 7 alginates lyases, AlyA, AlyB, AlyD, and AlyE, from V. splendidus 12B01. The four lyases were found to have optimal activity between pH 7.5 and 8.5 and at 20 to 25°C, consistent with their use in a marine environment. AlyA, AlyB, AlyD, and AlyE were found to exhibit a turnover number (k cat ) for alginate of 0.60 ؎ 0.02 s ؊1 , 3.7 ؎ 0.3 s ؊1 , 4.5 ؎ 0.5 s ؊1 , and 7.1 ؎ 0.2 s ؊1 , respectively. The K m values of AlyA, AlyB, AlyD, and AlyE toward alginate were 36 ؎ 7 M, 22 ؎ 5 M, 60 ؎ 2 M, and 123 ؎ 6 M, respectively. AlyA and AlyB were found principally to cleave the ␤-1,4 bonds between ␤-D-mannuronate and ␣-L-guluronate and subunits; AlyD and AlyE were found to principally cleave the ␣-1,4 bonds involving ␣-L-guluronate subunits. The four alginate lyases degrade alginate into longer chains of oligomers. Alginate is an abundant polysaccharide found within the cell wall of brown seaweeds (1) and comprises approximately 40% of the dry weight of the plant (2). Alginate is a copolymer consisting of the 1,4-linked epimers ␣-L-guluronate (G) and ␤-Dmannuronate (M). The individual monomeric subunits are organized in short stretches of polyguluronate (polyG), polymannuronate (polyM), or alternating sequences of mannuronate and guluronate (polyMG) (3). Alginate recently has been considered as a source for bioenergy, as it offers several potential advantages over terrestrial biomass. In particular, the farming of algae does not impinge on arable land; hence, it avoids the conflict between food and energy (4, 5). Algae also display higher growth rates than terrestrial plants (6). Finally, algae lack crystalline cellulose and lignin (7,8), bypassing a key obstacle to biofuel production.Alginate lyases are enzymes that degrade alginate through the ␤-elimination of the glycosidic bond into smaller oligomers. These enzymes can be classified based on the specific dyad G-G (EC 4.2.2.11) (9, 10), M-M (EC 4.2.2.3) (11), and M-G/G-M (12) bonds that they cleave. Additionally, alginate lyases can have either endocleaving or exocleaving specificity, with the majority of alginate lyases having an endocleaving preference (3). Along with the substrate specificity, alginate lyases also are characterized by their structure, termed polysaccharide lyase (PL) families. A total of seven PL families have been identified: PL5, PL6, PL7, PL14, PL15, PL17, and PL18 (13, 14). The most prevalent of these families is PL7 (Carbohydrate Active Enzymes database; http://www .cazy.org). The PL7 domain contains a ␤-jelly roll that consists of ␤-sheets in an antiparallel, adjacent barrel forming a cleft (12). This cleft contains three adjacent ␤-sheets that contain the catalytic residues (15). PL7 contains enzymes...

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“…To isolate alginate-degrading bacteria, 1.0% sodium alginate solution was poured over specified 100 cm 2 areas of soil once a week for three months [19] . In addition, another area of the soil was sampled directly (without alginate solution).…”

Section: Screening Of Alginate Lyase-producing Bacteriamentioning

confidence: 99%

“…lginate, a linear unbranched heteropolysaccharide, is composed of (1-4)-linked β-D-mannuronate (M) and α-L-guluronate (G), which are arranged by glycosidic bonds as a polyM block, a polyG block, and a random polyMG block [1,2] . The relative amount and distribution of these two residues vary with species and growth conditions [3] .…”

Section: Introductionmentioning

confidence: 99%

Screening of Alginate Lyase-Producing Bacteria and Optimization of Media Compositions for Extracellular Alginate Lyase Production

Tavafi¹,

Ali²,

Ghadam³

et al. 2017

IBJ

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Background: Alginate is a linear polysaccharide consisting of guluronate (polyG) and mannuronate (polyM) subunits. Methods: In the initial screening of alginate-degrading bacteria from soil, 10 isolates were able to grow on minimal medium containing alginate. The optimization of cell growth and alginate lyase (algL) production was carried out by the addition of 0.8% alginate and 0.2-0.3 M NaCl to the culture medium. Of 10 isolates, one was selected based on its fast growth rate on minimal 9 medium containing 0.4% sodium alginate. The selected bacterium, identified based on morphological and biochemical characteristics, as well as 16S rDNA sequence data, was confirmed to be an isolate belonging to the genus Bacillus and designated as Bacillus sp. TAG8. Results: The results showed the ability of Bacillus sp. TAG8 in utilizing alginate as a sole carbon source. Bacillus sp. TAG8 growth and algL production were augmented with an increase in sodium alginate concentration and also by the addition of 0.2-0.3 M NaCl. Molecular analysis of TAG8 algL gene showed 99% sequence identity with algL of Pseudomonas aeruginosa PAO1. The algL produced by Bacillus sp. TAG8 cleaved both polyM and polyG blocks in alginate molecule, as well as acetylated alginate residues, confirming the bifunctionality of the isolated lyase. Conclusion: The identification of novel algL genes from microbial communities constitutes a new approach for exploring lyases with specific activity against bacterial alginates and may thus contribute to the eradication of persistent biofilms from clinical samples.

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Purification and characterization of a new alginate lyase from a marine bacterium Vibrio sp.

Abstract: The purpose of this study is to find a polyG-preference alginate lyase for the saccharification of alginate combined with our polyM-preference alginate lyases.

Cited by 42 publications

References 13 publications

Purification and characterization of alginate lyase from locally isolated marinePseudomonas stutzeriMSEA04

Purification and characterization of alginate lyase from locally isolated marinePseudomonas stutzeriMSEA04

Alginate Lyases from Alginate-Degrading Vibrio splendidus 12B01 Are Endolytic

Screening of Alginate Lyase-Producing Bacteria and Optimization of Media Compositions for Extracellular Alginate Lyase Production

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