TY - JOUR
T1 - Biochemical characterization of LysVpKK5 endolysin from a marine vibriophage
AU - Melo-López, Felipe Neri
AU - Zermeño-Cervantes, Lina Angélica
AU - Barraza, Aarón
AU - Loera-Muro, Abraham
AU - Cardona-Félix, César Salvador
N1 - Funding Information:
Funding for research was provided by Consejo Nacional de Ciencia y Tecnología from México ( CONACyT México , grant 549477). LAZC acknowledges Doctoral fellowship provided by CONACYT No. 239000 and BEIFI-IPN No. 2466.
Funding Information:
Funding for research was provided by Consejo Nacional de Ciencia y Tecnolog?a from M?xico (CONACyT M?xico, grant 549477). LAZC acknowledges Doctoral fellowship provided by CONACYT No. 239000 and BEIFI-IPN No. 2466.
Publisher Copyright:
© 2021 Elsevier Inc.
PY - 2021/12
Y1 - 2021/12
N2 - Endolysins have been proposed as a potential antibacterial alternative for aquaculture, especially against Vibrio; the bacterial-agents that most frequently cause disease. Although multiple marine vibriophages have been characterized to date, research on vibriophage endolysins is recent. In this study, biochemical characterization of LysVpKK5 endolysin encoded by Vibrio parahaemolyticus-infecting VpKK5 phage was performed. In silico analysis revealed that LysVpKK5 possesses a conserved amidase_2 domain with a zinc-binding motif of high structural similarity to T7 lysozyme (RMSD = 0.107 Å). Contrary to expectations, the activity was inhibited with Zn2+ and was improved with other divalent cations, especially Ca2+. It showed optimal muralytic activity at pH 10, and curiously, no lytic activity at pH ≤ 7 was recorded. As for the thermal stability test, the optimal activity was recorded at 30 °C; the higher residual activity was recorded at 4 °C, and was lost at ≥ 50 °C. On the other hand, increasing NaCl concentrations reduced the activity gradually; the optimal activity was recorded at 50 mM NaCl. On the other hand, the enzymatic activity at 0.5 M NaCl was approx 30% and of approx 50% in seawater. LysVpKK5 endolysin exhibited a higher activity on V. parahaemolyticus ATCC-17802 strain, in comparison with AHPND + strains.
AB - Endolysins have been proposed as a potential antibacterial alternative for aquaculture, especially against Vibrio; the bacterial-agents that most frequently cause disease. Although multiple marine vibriophages have been characterized to date, research on vibriophage endolysins is recent. In this study, biochemical characterization of LysVpKK5 endolysin encoded by Vibrio parahaemolyticus-infecting VpKK5 phage was performed. In silico analysis revealed that LysVpKK5 possesses a conserved amidase_2 domain with a zinc-binding motif of high structural similarity to T7 lysozyme (RMSD = 0.107 Å). Contrary to expectations, the activity was inhibited with Zn2+ and was improved with other divalent cations, especially Ca2+. It showed optimal muralytic activity at pH 10, and curiously, no lytic activity at pH ≤ 7 was recorded. As for the thermal stability test, the optimal activity was recorded at 30 °C; the higher residual activity was recorded at 4 °C, and was lost at ≥ 50 °C. On the other hand, increasing NaCl concentrations reduced the activity gradually; the optimal activity was recorded at 50 mM NaCl. On the other hand, the enzymatic activity at 0.5 M NaCl was approx 30% and of approx 50% in seawater. LysVpKK5 endolysin exhibited a higher activity on V. parahaemolyticus ATCC-17802 strain, in comparison with AHPND + strains.
KW - Antimicrobials
KW - Aquaculture
KW - Bacteriophage
KW - Recombinant protein
KW - Shellfish
UR - http://www.scopus.com/inward/record.url?scp=85114746162&partnerID=8YFLogxK
U2 - 10.1016/j.pep.2021.105971
DO - 10.1016/j.pep.2021.105971
M3 - Artículo
C2 - 34508857
AN - SCOPUS:85114746162
SN - 1046-5928
VL - 188
JO - Protein Expression and Purification
JF - Protein Expression and Purification
M1 - 105971
ER -