Discovery of two novel β-glucosidases from an Amazon soil metagenomic library.
An Amazon soil microbial community metagenomic fosmid library was functionally screened for β-glucosidase activity. Contig analysis of positive clones revealed the presence of two ORFs encoding novel β-glucosidases, AmBGL17 and AmBGL18, from the GH3 and GH1 families, respectively. Both AmBGL17 and AmBGL18 were functionally identified as β-glucosidases. The enzymatic activity of AmBGL17 was further characterized. AmBGL17 was tested with different substrates and showed highest activity on pNPβG substrate with an optimum temperature of 45 °C and an optimum pH of 6. AmBGL17 showed a Vmax of 116 mM s(-1) and Km of 0.30 ± 0.017 mM. This is the first report of β-glucosidases from an Amazon soil microbial community using a metagenomic approach.
Genomic Sciences and Biotechnology Program, Universidade Católica de Brasília, Brasília, Brazil.
Jessica C
JC
Bergmann
Ohana Yonara A
OY
Costa
John M
JM
Gladden
Steven
S
Singer
Richard
R
Heins
Patrik
P
D'haeseleer
Blake A
BA
Simmons
Betania F
BF
Quirino
D044602:Cellulases / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism*; D004795:Enzyme Stability; D015723:Gene Library; D005820:Genetic Testing; D006863:Hydrogen-Ion Concentration; D056186:Metagenomics; D008969:Molecular Sequence Data; D017422:Sequence Analysis, DNA; D012988:Soil Microbiology; D013379:Substrate Specificity; D013696:Temperature
D044602:Cellulases
GH1; GH3; beta-glucosidase; functional screen; metagenomic library