Purification and characterization of extracellular lipases from Pseudomonas monteilii TKU009 by the use of soybeans as the substrate.
A lipase-producing bacterium was isolated and identified as Pseudomonas monteilii TKU009. A lipase (F2) and lipase-like materials (F1) were purified from the culture supernatant of P. monteilii TKU009 with soybean powder as the sole carbon/nitrogen source. The molecular mass of F1 and F2 was estimated to be 44 kDa by SDS-PAGE and gel filtration. The optimum pH, optimum temperature, and pH and thermal stabilities of F2 were 7, 40 degrees C, 8-11, and 50 degrees C; and of F1 were 6, 40 degrees C, 6-7, and 50 degrees C, respectively. F2 was completely inhibited by EDTA and slightly by Mg(2+), Fe(2+), Mn(2+), and SDS. F1 was completely inhibited by EDTA and Fe(2+) and strongly by Zn(2+), Mn(2+), Ca(2+), Mg(2+), and SDS. The activities of both the enzymes were enhanced by the addition of non-ionic surfactants Triton X-100 and Tween 40, especially for F1. F2 preferably acted on substrates with a long chain (C10-C18) of fatty acids, while F1 showed a broad spectrum on those with chain length of C4-C18. The marked activity of F2 in organic solvents makes it an ideal choice for application in a water-restricted medium including organic synthesis.
Wang
San-Lang
SL
Graduate Institute of Life Sciences, Tamkang University, Tamsui, 251, Taiwan. sabulo@mail.tku.edu.tw
Lin
Yu-Ting
YT
Liang
Tzu-Wen
TW
Chio
Sau-Hua
SH
Ming
Li-June
LJ
Wu
Pei-Chen
PC
D001426:Bacterial Proteins / Q000737:chemistry* / Q000302:isolation & purification* / Q000378:metabolism; D003470:Culture Media / Q000378:metabolism; D004795:Enzyme Stability; D005110:Extracellular Space / Q000737:chemistry / Q000201:enzymology*; D008049:Lipase / Q000737:chemistry* / Q000302:isolation & purification* / Q000378:metabolism; D008970:Molecular Weight; D010802:Phylogeny; D011549:Pseudomonas / Q000737:chemistry / Q000145:classification / Q000201:enzymology* / Q000302:isolation & purification; D012988:Soil Microbiology; D013025:Glycine max / Q000378:metabolism*; D013379:Substrate Specificity
D001426:Bacterial Proteins; D003470:Culture Media; D008049:Lipase