TY - JOUR
T1 - Galactofuranosidase from JHA 19 Streptomyces sp.
T2 - subcloning and biochemical characterization
AU - Seničar, Mateja
AU - Legentil, Laurent
AU - Ferrières, Vincent
AU - Eliseeva, Svetlana V.
AU - Petoud, Stéphane
AU - Takegawa, Kaoru
AU - Lafite, Pierre
AU - Daniellou, Richard
N1 - Funding Information:
This work was supported by the APR IR NeoLect project from the Région Centre-Val de Loire, France . S. V. E. and S. P. acknowledge support from La Ligue Régionale contre le Cancet and from Institut National de la Santé et de la Recherche Médicale (INSERM).
Publisher Copyright:
© 2019 Elsevier Ltd
PY - 2019/7/1
Y1 - 2019/7/1
N2 - Despite the crucial role of the rare galactofuranose (Galf) in many pathogenic micro-organisms and our increased knowledge of its metabolism, there is still a lack of recombinant and efficient galactofuranoside hydrolase available for chemo-enzymatic synthetic purposes of specific galactofuranosyl-conjugates. Subcloning of the Galf-ase from JHA 19 Streptomyces sp. and its further overexpression lead us to the production of this enzyme with a yield of 0.5 mg/L of culture. It exhibits substrate specificity exclusively towards pNP β-D-Galf, giving a KM value of 250 μM, and the highest enzymatic efficiency ever observed of 14 mM−1 s−1. It proved to be stable to temperature up to 60 °C and to at least 4 freeze-thaw's cycles. Thus, Galf-ase demonstrated to be an efficient and stable biocatalyst with greatly improved specificity toward the galactofuranosyl entity, thus paving the way to the further development of transglycosylation and thioligation reactions.
AB - Despite the crucial role of the rare galactofuranose (Galf) in many pathogenic micro-organisms and our increased knowledge of its metabolism, there is still a lack of recombinant and efficient galactofuranoside hydrolase available for chemo-enzymatic synthetic purposes of specific galactofuranosyl-conjugates. Subcloning of the Galf-ase from JHA 19 Streptomyces sp. and its further overexpression lead us to the production of this enzyme with a yield of 0.5 mg/L of culture. It exhibits substrate specificity exclusively towards pNP β-D-Galf, giving a KM value of 250 μM, and the highest enzymatic efficiency ever observed of 14 mM−1 s−1. It proved to be stable to temperature up to 60 °C and to at least 4 freeze-thaw's cycles. Thus, Galf-ase demonstrated to be an efficient and stable biocatalyst with greatly improved specificity toward the galactofuranosyl entity, thus paving the way to the further development of transglycosylation and thioligation reactions.
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U2 - 10.1016/j.carres.2019.05.011
DO - 10.1016/j.carres.2019.05.011
M3 - Article
C2 - 31174175
AN - SCOPUS:85066476456
SN - 0008-6215
VL - 480
SP - 35
EP - 41
JO - Carbohydrate Research
JF - Carbohydrate Research
ER -