Protocols in Current Issue
    Chromatographic Assays for the Enzymatic Degradation of Chitin
    Authors:  Sophanit Mekasha, Tina R. Tuveng, Gustav Vaaje-Kolstad and Vincent G. H. Eijsink, date: 05/05/2021, view: 297, Q&A: 0

    Chitin is an insoluble linear polymer of β(1→4)-linked N-acetylglucosamine. Enzymatic cleavage of chitin chains can be achieved using hydrolytic enzymes, called chitinases, and/or oxidative enzymes, called lytic polysaccharide monooxygenases (LPMOs). These two groups of enzymes have different modes of action and yield different product

    Quantitative Characterization of the Amount and Length of (1,3)-β-D-glucan for Functional and Mechanistic Analysis of Fungal (1,3)-β-D-glucan Synthase
    Authors:  Abhishek Chhetri, Anna Loksztejn and Kenichi Yokoyama, date: 04/20/2021, view: 817, Q&A: 0

    (1,3)-β-d-Glucan synthase (GS) is an essential enzyme for fungal cell wall biosynthesis that catalyzes the synthesis of (1,3)-β-d-glucan, a major and vital component of the cell wall. GS is a proven target of antifungal antibiotics including FDA-approved echinocandin derivatives; however, the function and mechanism of GS remain largely

    Negative Ion Mode nanoLC-ESI-MS/MS Analyses of Permethylated Sulfated Glycans
    Authors:  Shin-Yi Yu, Chu-Wen Cheng and Kay-Hooi Khoo, date: 05/20/2020, view: 1720, Q&A: 0
    [Abstract] We have developed enabling techniques for sulfoglycomics based on MALDI-MS mapping and MS/MS sequencing of permethylated sulfated glycans. We then extended further the analytical workflow to C18 reverse phase (RP)-nanoLC-nanoESI-MS/MS analyses of permethylated sulfated glycans in the negative ion mode. The advantages are that extra sulfates on ...
    Permethylation and Microfractionation of Sulfated Glycans for MS Analysis
    Authors:  Shin-Yi Yu, Sergei Snovida and Kay-Hooi Khoo, date: 05/20/2020, view: 1754, Q&A: 0
    [Abstract] Sulfated glycans are barely detectable in routine mass spectrometry (MS)-based glycomic analysis due to ion suppression by the significantly more abundant neutral glycans in the positive ion mode, and sialylated non-sulfated glycans in the negative ion mode, respectively. Nevertheless, the negative charge imparted by sulfate can be advantageous ...
    Biofilm Formation Assay in Pseudomonas syringae
    Authors:  Xiaolong Shao, Yingpeng Xie, Yingchao Zhang and Xin Deng, date: 05/20/2019, view: 6899, Q&A: 1
    [Abstract] Pseudomonas syringae is a model plant pathogen that infects more than 50 plant species worldwide, thus leading to significant yield loss. Pseudomonas biofilm always adheres to the surfaces of medical devices or host cells, thereby contributing to infection. Biofilm formation can be visualized on numerous matrixes, including ...
    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: 4159, 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 ...
    Determination of Intracellular Osmolytes in Cyanobacterial Cells
    Authors:  Xiaoming Tan, Kuo Song, Cuncun Qiao and Xuefeng Lu, date: 04/20/2018, view: 3753, Q&A: 0
    [Abstract] Most of the cyanobacteria accumulate osmolytes including sucrose, glucosylglycerol, in their cells in response to salt stress. Here we describe a protocol of our laboratory for extraction and quantification of cyanobacterial intracellular sucrose and glucosylglycerol. We have confirmed this protocol was applicable to at least four kinds of ...
    Preparation of the Partially Methylated Alditol Acetates Derived from CS Tetrasaccharides Containing Galactose for the Gas Chromatography/Mass Spectrometry Analysis
    Authors:  Kyohei Higashi and Toshihiko Toida, date: 11/05/2017, view: 7005, Q&A: 0
    [Abstract] Chondroitin sulfate (CS), a member of the glycosaminoglycan (GAG) family of carbohydrates, is composed of linear, sulfated repeating disaccharide sequences of N-acetyl-D-galactosamine (GalNAc) and glucuronic acid (GlcA). Recently, a keratan sulfate (KS) disaccharide [GlcNAc6S(β1-3)Galactose(β1-]-branched CS-E was identified ...
    Observation of Pneumococcal Phase Variation in Colony Morphology
    Authors:  Jing Li, Juanjuan Wang, Fangfang Jiao and Jing-Ren Zhang, date: 08/05/2017, view: 7092, 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 ...
    Separation and Purification of Glycosaminoglycans (GAGs) from Caenorhabditis elegans
    Authors:  Tabea Dierker and Lena Kjellén, date: 08/05/2017, view: 8749, Q&A: 1
    [Abstract] The nematode Caenorhabditis elegans is a popular model organism for studies of developmental biology, neurology, ageing and other fields of basic research. Because many developmental processes are regulated by glycosaminoglyans (GAGs) on cell surfaces and in the extracellular matrix, methods to isolate and analyze C. elegans GAGs ...

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