|Formulation:||In 3.2 M ammonium sulphate|
|Stability:||> 4 years at 4oC|
|Enzyme Activity:||Alginate Lyase|
|Synonyms:||poly(beta-D-mannuronate) lyase; poly[(1,4)-beta-D-mannuronide] lyase|
|Concentration:||Supplied at ~ 2,500 U/mL|
|Expression:||Recombinant from Sphingomonas sp.|
|Specificity:||endo-acting β-elimination cleavage of the polysaccharide, alginate.|
|Specific Activity:||~ 120 U/mg (40oC, pH 7.2 on sodium alginate)|
|Unit Definition:||One Unit of alginate lyase activity is defined as the amount of enzyme required to produce an increase in absorbance of 1.0 per minute at 235 nm in Tris.HCl buffer (100mM), pH 7.2 at 40oC.|
|Application examples:||Utilised in research, biochemical assays and in vitro binding studies and as a biocatalyst for saccharification of alginate.|
High purity recombinant Alginate lyase (Sphingomonas sp.) for use in research, biochemical enzyme assays and in vitro diagnostic analysis.
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Production of bioethanol from brown algae.
Ravanal, M. C., Camus, C., Buschmann, A. H., Gimpel, J., Olivera-Nappa, Á., Salazar, O. & Lienqueo, M. E. (2019). “Advances in Feedstock Conversion Technologies for Alternative Fuels and Bioproducts”, Woodhead Publishing, pp. 69-88.
As an alternative to the use of edible crops as raw materials or the use of lignocellulosic materials for production of the second-generation biofuels, macroalgae biomass has potential as a sugar resource for the production of third-generation biofuels. As higher amounts of biomass are required for this endeavor, commercial cultivation of brown algae is necessary to avoid overexploitation of natural populations. The macroalgae species contain different kinds of polysaccharides and pretreatment is necessary to make them available using different strategies. The most abundant sugars in brown algae are alginate, mannitol, and glucan; whereby the degradation of these polysaccharides requires specific enzymes for the release of monosaccharides. Monosaccharides are most efficiently fermented into ethanol by engineered microorganisms, but as recently demonstrated, both Saccharomyces cerevisiae and Escherichia coli strains have this capability also. Saccharification and fermentation steps can be carried out in different configurations. This chapter presents a review of each step of the process for bioethanol production from brown algae.Hide Abstract
DMTMM-mediated amidation of alginate oligosaccharides aimed at modulating their interaction with proteins.
Labre, F., Mathieu, S., Chaud, P., Morvan, P. Y., Vallée, R., Helbert, W. & Fort, S. (2018). Carbohydrate Polymers, 184, 427-434.
Alginate oligosaccharides (AOS) with a weight average molecular weight of 5 kDa were efficiently amidated with amino acids and carbohydrates in aqueous media in the presence of 4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholinium chloride (DMTMM). Here, alanine, leucine, serine, as well as mannose and rhamnose, were amidated at high yields with a good control of the degree of substitution (DS). Amino acid- and carbohydrate-grafted AOS showed improved stability against degradation by alginate lyases having different specificities. This enzyme resistance was correlated with the DS: hydrolysis was reduced by 60 to 70% for low DS (0.1), whereas AOS with DS ranging from 0.4 to 0.6 remained unhydrolyzed. Competitive inhibition assays demonstrated multivalent binding of mannose-amidated AOS to concanavalin A lectin. A 178-fold affinity enhancement was observed for AOSMan-0.38 (DS 0.38) over α-methyl-mannoside with an IC50 of 5.6 µM, lending further evidence for the promising potential of AOS as multivalent scaffolds.Hide Abstract
Effect of alginate size, mannuronic/guluronic acid content and pH on particle size, thermodynamics and composition of complexes with β-lactoglobulin.
Stender, E. G., Khan, S., Ipsen, R., Madsen, F., Hägglund, P., Hachem, M. A., Almdal, K., Westh, P. & Svensson, B. (2018). Food Hydrocolloids, 75, 157-163.
Alginate is an anionic polysaccharide capable of forming insoluble particles with proteins. Hence, alginate has potential as a protein carrier. However, the role of physical properties of the polysaccharide, such as degree of polymerization (DPn) and mannuronic/guluronic acid ratio, remains to be fully explored. Particle formation of a high and a low molar mass alginate (ALG) with β-lactoglobulin (BLG) at pH 2-8 depends on the average DPn (HMW-ALG: 1.59·103; LMW-ALG: 0.23·103) and the mannuronic/guluronic acid ratio (1.0; 0.6) as supported by using ManA6 and GulA6 as models. Dynamic light scattering (DLS) showed that particles of BLG with either of the two ALGs have essentially the same hydrodynamic diameter (DH) at pH 3 and 2, while at pH 4 particles of LMW-ALG/BLG have larger DH than of HMW-ALG/BLG. At pH 5-8 no significant particle formation was observed. ManA6 did not form insoluble particles at pH 2-8, while GulA6 formed insoluble particles, albeit only at pH 4. Kd was approximately 10-fold higher for LMW-ALG/BLG than HMW-ALG/BLG and 3 orders of magnitude higher for an alginate trisaccharide/BLG complexation as determined by isothermal titration calorimetry (ITC). The alginate trisaccharide did not form insoluble particles with BLG at pH 3 and 4, though interaction still occurred. δHapp and molar stoichiometry of BLG in the complexes with the two ALGs differed by a factor of 7, as did their DPn, which thus affected the interaction strength, but not the BLG content. At pH 4 the BLG content doubled in the particle due to BLG dimerization. The findings emphasize the importance of DPn, mannuronic/guluronic acid ratio and pH in formulations containing alginate/whey protein particles.Hide Abstract
Jonathan, M. C., Bosch, G., Schols, H. A. & Gruppen, H. (2013). Journal of Agricultural and Food Chemistry, 61(3), 553-560.
This research aimed to develop a method for analyzing specific alginate oligosaccharides (AOS) in a complex matrix such as pig feces. The data obtained were used to study alginate degradation by the microbiota in the large intestine during adaptation, including the individual variation between pigs. A method using an UHPLC system with an ethylene bridged hybrid (BEH) amide column coupled with MSn detection was able to distinguish saturated and unsaturated AOS with DP 2–10. Isomers of unsaturated trimer and tetramer could be separated and annotated. In the feces, saturated and unsaturated AOS were present. The presence of unsaturated AOS indicates that the microbiota produced alginate lyase. The microbiota utilized unsaturated AOS more than saturated AOS. The results also suggested that guluronic acid at the reducing end of AOS inhibits the utilization by microbiota during the first weeks of adaptation. After adaptation, the microbiota was able to utilize a broader range of AOS.Hide Abstract