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Generation of Giant Unilamellar Vesicles (GUVs) Using Polyacrylamide Gels

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Determination of Storage (Starch/Glycogen) and Total Saccharides Content in Algae and Cyanobacteria by a Phenol-Sulfuric Acid Method

Authors: Tomáš Zavřel, Petra Očenášová, Maria A. Sinetova and Jan Červený, date: 08/05/2018, view: 5833, Q&A: 0

[Abstract] This is a protocol for quantitative determination of storage and total carbohydrates in algae and cyanobacteria. The protocol is simple, fast and sensitive and it requires only few standard chemicals. Great advantage of this protocol is that both storage and total saccharides can be determined in the cellular pellets that were already used for ...

Enzymatic Activity Assay for Invertase in Synechocystis Cells

Authors: Xiaoming Tan, Kuo Song and Xuefeng Lu, date: 05/20/2018, view: 4571, Q&A: 0

[Abstract] Invertase can catalyze the hydrolysis of sucrose, and is widely distributed in cells of cyanobacteria and plants. Being responsible for the first step for sucrose metabolism, invertase plays important physiological roles and its enzymatic activity is frequently needed to be determined. All the methods for determination of the invertase activity ...

Glycogen and Extracellular Glucose Estimation from Cyanobacteria Synechocystis sp. PCC 6803

Authors: Md. Rezaul Islam Khan, Yushu Wang, Shajia Afrin, Lin He and Gang Ma, date: 05/05/2018, view: 3906, Q&A: 0

[Abstract] Cyanobacteria, which have the extraordinary ability to grow using sunlight and carbon dioxide, are emerging as a green host to produce value-added products. Exploitation of this highly promising host to make products may depend on the ability to modulate the glucose metabolic pathway; it is the key metabolic pathway that generates intermediates ...

Laminarin Quantification in Microalgae with Enzymes from Marine Microbes

Authors: Stefan Becker and Jan-Hendrik Hehemann, date: 04/20/2018, view: 4466, Q&A: 0

[Abstract] 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 ...

Bacterial Cell Wall Precursor Phosphatase Assays Using Thin-layer Chromatography (TLC) and High Pressure Liquid Chromatography (HPLC)

Authors: Manuel Pazos, Christian Otten and Waldemar Vollmer, date: 03/20/2018, view: 3934, Q&A: 0

[Abstract] Peptidoglycan encases the bacterial cytoplasmic membrane to protect the cell from lysis due to the turgor. The final steps of peptidoglycan synthesis require a membrane-anchored substrate called lipid II, in which the peptidoglycan subunit is linked to the carrier lipid undecaprenol via a pyrophosphate moiety. Lipid II is the target of ...

Observation of Pneumococcal Phase Variation in Colony Morphology

Authors: Jing Li, Juanjuan Wang, Fangfang Jiao and Jing-Ren Zhang, date: 08/05/2017, view: 6746, Q&A: 0

[Abstract] Streptococcus pneumoniae (pneumococcus) is an important human pathogen that causes pneumonia, meningitis, sepsis, and otitis media. This bacterium normally resides in the nasopharynx as a commensal, but sometimes disseminates to sterile sites of humans and causes local or systemic inflammation. This biphasic behavior of S. pneumoniae ...

Digestion of Peptidoglycan and Analysis of Soluble Fragments

Authors: Ryan E. Schaub and Joseph P. Dillard, date: 08/05/2017, view: 8982, Q&A: 0

[Abstract] Peptidoglycan (murein) is a vital component of the cell wall of nearly all bacteria, composed of sugars linked by short peptides. This protocol describes the purification of macromolecular peptidoglycan from cultured bacteria and the analysis of enzyme-digested peptidoglycan fragments using high performance liquid chromatography (HPLC). Digested ...

Measurement of the Galactanase Activity of the GanB Galactanase Protein from Bacillus subtilis

Author: Hildegard Watzlawick, date: 04/05/2017, view: 5872, Q&A: 0

[Abstract] The activity of the endo-β-1,4-galactanase GanB from B. subtilis on the high molecular weight β-1,4-galactan was determined quantitatively by the measurement of the increase of the reducing power or with the dyed substrate Azo-galactan. The generated degradation products were analyzed using thinlayer-chromatography (TLC) or ...

Chitin Extraction and Content Measurement in Magnaporthe oryzae

Authors: Xinyu Liu and Zhengguang Zhang, date: 03/05/2017, view: 6100, Q&A: 0

[Abstract] Chitin is a linear polysaccharide composed of β (1→4)-linked N-acetylglucosamine (GlcNAc) residues. In fungi, chitin is an important component of the cell wall. Here, we provide a protocol to measure the chitin content of fungal cells using Magnaporthe oryzae as an example.

Determination of Keto-deoxy-d-manno-8-octanoic acid (KDO) from Lipopolysaccharide of Escherichia coli

Authors: M. R. Sunayana and Manjula Reddy, date: 12/20/2015, view: 13869, Q&A: 0

[Abstract] 2-Keto-3-deoxy-octonate (KDO) is an essential constituent of lipopolysaccharide (LPS) that forms the outermost leaflet of Gram-negative bacterial outer membrane. LPS is mainly composed of lipid A, O-antigen and a core oligosaccharide. Two molecules of KDO are present per one molecule of LPS. A proper level of LPS is required to maintain the outer ...

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