Biotinylation of the rhizobial cyclic β-glucans and succinoglycans crucial for symbiosis with legumes.
The cyclic β-glucans and succinoglycans produced by rhizobia are required for nodulation during symbiosis with legume hosts. However, only gene deletion analyses have been used to investigate their biological importance. For future studies on the physiological activity of those during symbiosis, biochemical methods need to be developed with separate carbohydrate compounds. Here, we isolated and purified rhizobial cellular carbohydrates using various chromatographic methods. Purified cyclic β-glucans, cyclosophoraoses, were monofunctionalized with biotin using the following three steps: tosylation, azidation, and amination. The mono-6-amino-cyclosophoraoses were linked with biotinamidohexanoic acid N-hydroxysuccinimide ester. Succinoglycans and monomers were tagged with biotinamidocaproyl hydrazide at the reducing sugar via reductive amination. The resulting biotinylated rhizobial carbohydrates were characterized by Fourier transform infrared and nuclear magnetic resonance spectroscopy, matrix-assisted laser desorption/ionization time-of-flight mass spectroscopy, and electrospray ionization mass spectrometry. The resulting neoglycoconjugates can be used as solid probes to study putative plant receptors and for non-invasive imaging for in vivo tracing.
Department of Bioscience and Biotechnology, Bio/Molecular Informatics Center (BMIC) & Institute for Ubiquitous Information Technology and Applications (CBRU), Konkuk University, 1 Hwayang-dong Gwangjin-gu, Seoul 143-701, Republic of Korea.
Eunae
E
Cho
NaraeBiotech, Incorporation R&D Center, 177, Dangha-ri, Bongdam-eup, Hwaseong-si, Gyeonggi-do 445-892, Republic of Korea.
Chanho
C
Kwon
Department of Bioanalysis, Seoul Medical Science Institute, Seoul 140-809, Republic of Korea.
Sanghoo
S
Lee
Department of Bioscience and Biotechnology, Bio/Molecular Informatics Center (BMIC) & Institute for Ubiquitous Information Technology and Applications (CBRU), Konkuk University, 1 Hwayang-dong Gwangjin-gu, Seoul 143-701, Republic of Korea.
Muhammad Nazir
MN
Tahir
Department of Applied Chemistry, Dongduk Women's University, Seoul 136-714, Republic of Korea.
Seyeon
S
Park
Department of Bioscience and Biotechnology, Bio/Molecular Informatics Center (BMIC) & Institute for Ubiquitous Information Technology and Applications (CBRU), Konkuk University, 1 Hwayang-dong Gwangjin-gu, Seoul 143-701, Republic of Korea. Electronic address: shjung@konkuk.ac.kr.
Seunho
S
Jung
D000586:Amination; D001386:Azides / Q000378:metabolism; D019921:Biotinylation; D007887:Fabaceae / Q000382:microbiology*; D011135:Polysaccharides, Bacterial / Q000378:metabolism*; D012231:Rhizobium / Q000378:metabolism* / Q000502:physiology; D013559:Symbiosis; D047071:beta-Glucans / Q000378:metabolism*
D001386:Azides; D011135:Polysaccharides, Bacterial; D047071:beta-Glucans; C056779:succinoglycan
Biotinylation; Cyclosophoraose; Rhizobiaceae; Succinoglycan; Symbiosis