TY - JOUR
T1 - Structural and functional characterization of a novel lipolytic enzyme from a Brazilian Cerrado soil metagenomic library
AU - Istvan, Paula
AU - Souza, Amanda Araújo
AU - Garay, Aisel Valle
AU - dos Santos, Debora Farage Knupp
AU - de Oliveira, Gideane Mendes
AU - Santana, Renata Henrique
AU - Lopes, Fabyano Alvares Cardoso
AU - de Freitas, Sonia Maria
AU - Barbosa, João Alexandre Ribeiro Gonçalves
AU - Krüger, Ricardo Henrique
N1 - Publisher Copyright:
© 2018, Springer Nature B.V.
PY - 2018/10/1
Y1 - 2018/10/1
N2 - Objective: To isolate putative lipase enzymes by screening a Cerrado soil metagenomic library with novel features. Results: Of 6720 clones evaluated, Clone W (10,000 bp) presented lipolytic activity and four predicted coding sequences, one of them LipW. Characterization of a predicted esterase/lipase, LipW, showed 28% sequence identity with an arylesterase from Pseudomonas fluorescens (pdb|3HEA) from protein database (PDB). Phylogenetic analysis showed LipW clustered with family V lipases; however, LipW was clustered in different subclade belonged to family V, suggesting a different subgroup of family V. In addition, LipW presented a difference in family V GH motif, a glycine replaced by a serine in GH motif. Estimated molecular weight and stokes radius values of LipW were 29,338.67–29,411.98 Da and 2.58–2.83 nm, respectively. Optimal enzyme activity was observed at pH 9.0–9.5 and at 40 °C. Circular dichroism analysis estimated secondary structures percentages as approximately 45% α-helix and 15% β-sheet, consistent with the 3D structure predicted by homology. Conclusion: Our results demonstrate the isolation of novel family V lipolytic enzyme with biotechnological applications from a metagenomic library.
AB - Objective: To isolate putative lipase enzymes by screening a Cerrado soil metagenomic library with novel features. Results: Of 6720 clones evaluated, Clone W (10,000 bp) presented lipolytic activity and four predicted coding sequences, one of them LipW. Characterization of a predicted esterase/lipase, LipW, showed 28% sequence identity with an arylesterase from Pseudomonas fluorescens (pdb|3HEA) from protein database (PDB). Phylogenetic analysis showed LipW clustered with family V lipases; however, LipW was clustered in different subclade belonged to family V, suggesting a different subgroup of family V. In addition, LipW presented a difference in family V GH motif, a glycine replaced by a serine in GH motif. Estimated molecular weight and stokes radius values of LipW were 29,338.67–29,411.98 Da and 2.58–2.83 nm, respectively. Optimal enzyme activity was observed at pH 9.0–9.5 and at 40 °C. Circular dichroism analysis estimated secondary structures percentages as approximately 45% α-helix and 15% β-sheet, consistent with the 3D structure predicted by homology. Conclusion: Our results demonstrate the isolation of novel family V lipolytic enzyme with biotechnological applications from a metagenomic library.
KW - Cerrado
KW - Lipolytic enzymes
KW - Metagenome
KW - Soil
UR - http://www.scopus.com/inward/record.url?scp=85050966947&partnerID=8YFLogxK
U2 - 10.1007/s10529-018-2598-0
DO - 10.1007/s10529-018-2598-0
M3 - Article
C2 - 30062528
AN - SCOPUS:85050966947
SN - 0141-5492
VL - 40
SP - 1395
EP - 1406
JO - Biotechnology Letters
JF - Biotechnology Letters
IS - 9-10
ER -