Identification and characterization of a bacterial hyaluronidase and its production in recombinant form.
Hyaluronidases (Hyals) are broadly used in medical applications to facilitate the dispersion and/or absorption of fluids or medications. This study reports the isolation, cloning, and industrial-scale recombinant production, purification and full characterization, including X-ray structure determination at 1.45 Å, of an extracellular Hyal from the nonpathogenic bacterium Streptomyces koganeiensis. The recombinant S. koganeiensis Hyal (rHyal_Sk) has a novel bacterial catalytic domain with high enzymatic activity, compared with commercially available Hyals, and is more thermostable and presents higher proteolytic resistance, with activity over a broad pH range. Moreover, rHyal_Sk exhibits remarkable substrate specificity for hyaluronic acid (HA) and poses no risk of animal cross-infection.
Local Unit Fidia Research Sud, Fidia Farmaceutici S.p.A., Noto, Italy.
Luciano
L
Messina
Laboratorio de Estudios Cristalográficos, Instituto Andaluz de Ciencias de la Tierra (Consejo Superior de Investigaciones Científicas-Universidad de Granada), Armilla, Spain.
Jose A
JA
Gavira
GENYO - Centre for Genomics and Oncological Research: Pfizer/University of Granada/Andalusian Regional Government, PTS Granada, Spain.
Salvatore
S
Pernagallo
NanoGetic S.L., PTS Granada, Armilla, Spain.
Juan D
JD
Unciti-Broceta
GENYO - Centre for Genomics and Oncological Research: Pfizer/University of Granada/Andalusian Regional Government, PTS Granada, Spain.|NanoGetic S.L., PTS Granada, Armilla, Spain.|Facultad de Farmacia, Universidad de Granada, Spain.
Rosario M
RM
Sanchez Martin
GENYO - Centre for Genomics and Oncological Research: Pfizer/University of Granada/Andalusian Regional Government, PTS Granada, Spain.|Facultad de Farmacia, Universidad de Granada, Spain.
Juan J
JJ
Diaz-Mochon
Local Unit Fidia Research Sud, Fidia Farmaceutici S.p.A., Noto, Italy.
Susanna
S
Vaccaro
Laboratorio de Estudios Cristalográficos, Instituto Andaluz de Ciencias de la Tierra (Consejo Superior de Investigaciones Científicas-Universidad de Granada), Armilla, Spain.
Mayte
M
Conejero-Muriel
Laboratorio de Estudios Cristalográficos, Instituto Andaluz de Ciencias de la Tierra (Consejo Superior de Investigaciones Científicas-Universidad de Granada), Armilla, Spain.
Estela
E
Pineda-Molina
Local Unit Fidia Research Sud, Fidia Farmaceutici S.p.A., Noto, Italy.
Salvatore
S
Caruso
Local Unit Fidia Research Sud, Fidia Farmaceutici S.p.A., Noto, Italy.
Luca
L
Musumeci
Local Unit Fidia Research Sud, Fidia Farmaceutici S.p.A., Noto, Italy.
Roberta
R
Di Pasquale
Primm S.r.l, Milano, Italy.
Angela
A
Pontillo
Primm S.r.l, Milano, Italy.
Francesca
F
Sincinelli
Fidia Farmaceutici S.p.A., Abano Terme, Italy.
Mauro
M
Pavan
Fidia Farmaceutici S.p.A., Abano Terme, Italy.
Cynthia
C
Secchieri
D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics; D004795:Enzyme Stability; D006821:Hyaluronoglucosaminidase / Q000737:chemistry* / Q000235:genetics; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics; D013302:Streptomyces / Q000201:enzymology* / Q000235:genetics
D001426:Bacterial Proteins; D011994:Recombinant Proteins; D006821:Hyaluronoglucosaminidase
Streptomyces koganeiensis; hyaluronic acid or hyaluronan; hyaluronidase