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  • Laminarin Quantification in Microalgae with Enzymes from Marine Microbes.

Laminarin Quantification in Microalgae with Enzymes from Marine Microbes.

Bio-protocol (2018-04-20)
Stefan Becker, Jan-Hendrik Hehemann
摘要

The marine beta-glucan laminarin is an abundant storage polysaccharide in microalgae. High production rates and rapid digestion by heterotrophic bacteria turn laminarin into an ideal carbon and energy source, and it is therefore a key player in the marine carbon cycle. As a main storage glucan laminarin also plays a central role in the energy metabolism of the microalgae (Percival and Ross, 1951; Myklestad, 1974; Painter, 1983). We take advantage of enzymes that digest laminarin selectively and can thereby quantify only this polysaccharide in environmental samples. These enzymes hydrolyze laminarin into glucose and oligosaccharides, which are measured with a standard reducing sugar assay to obtain the laminarin concentration. Prior to this assay, the three enzymes need to be produced via heterologous expression and purification. The assay can be used to monitor laminarin concentrations in environmental microalgae, which were concentrated from seawater by filtering, or in samples derived from algal lab cultures.

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Sigma-Aldrich
Triton X-100, laboratory grade
Sigma-Aldrich
脱氧胆酸钠, ≥97% (titration)
Sigma-Aldrich
D -(+)-葡萄糖, ≥99.5% (GC)
Sigma-Aldrich
DL-二硫苏糖醇, ≥98% (HPLC), ≥99.0% (titration)
Sigma-Aldrich
硫酸铵, for molecular biology, ≥99.0%
Sigma-Aldrich
Lysozyme 来源于鸡蛋白, lyophilized powder, protein ≥90 %, ≥40,000 units/mg protein
Sigma-Aldrich
柠檬酸钠二水合物, ≥99%, FG
Sigma-Aldrich
咪唑, for molecular biology, ≥99% (titration)
Sigma-Aldrich
甘油, BioXtra, ≥99% (GC)
Sigma-Aldrich
4-羟基苯肼, ≥97%
Sigma-Aldrich
昆布多糖 来源于掌状海带, polysaccharide substrate for laminarinase
Sigma-Aldrich
琼脂, ash 2.0-4.5%