Cloning, expression, purification, and characterization of a novel esterase from Lactobacillus plantarum.
Lactobacillus plantarum is an important lactic acid bacterium, usually found as natural inhabitant of food, such as fermented vegetables and meat products. However, little information about lactic acid bacteria, especially concerning L. plantarum, as a source of useful enzymes has been reported. The aim of this study was to clone, express in Escherichia coli, purify, and characterize an esterase from L. plantarum ATCC 8014. The esterase gene (1014 bp) was amplified and cloned in pET14b expression vector to express a His(6)-tagged protein in E. coli. Recombinant L. plantarum esterase was purified by Ni-NTA resin, presenting an apparent molecular mass of about 38 kDa. It presented highest activity at pH 6.0 and 40 degrees C. Also, it presented preference for p-nitrophenyl butyrate, but hydrolyzed more efficiently p-nitrophenyl acetate. Besides, this study shows, for the first time, CD data about secondary structure of an esterase from L. plantarum.
Brod
Fábio Cristiano Angonesi
FC
Department of Food Science and Technology, Universidade Federal de Santa Catarina, Florianópolis, Brazil.
Vernal
Javier
J
Bertoldo
Jean Borges
JB
Terenzi
Hernán
H
Arisi
Ana Carolina Maisonnave
AC
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism*; D002087:Butyrates / Q000378:metabolism; D003001:Cloning, Molecular; D004269:DNA, Bacterial; D004926:Escherichia coli / Q000235:genetics; D004950:Esterases / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism*; D005798:Genes, Bacterial; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D007700:Kinetics; D048191:Lactiplantibacillus plantarum / Q000201:enzymology* / Q000235:genetics; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009596:Nitrophenols / Q000378:metabolism; D010957:Plasmids; D017433:Protein Structure, Secondary / Q000235:genetics; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D013056:Spectrophotometry, Ultraviolet; D013379:Substrate Specificity; D013696:Temperature
D001426:Bacterial Proteins; D002087:Butyrates; D004269:DNA, Bacterial; D009596:Nitrophenols; D011994:Recombinant Proteins; C033592:4-nitrophenyl butyrate; C008642:4-nitrophenyl acetate; D004950:Esterases