The improvement of lipase secretion and stability by addition of inert compounds into Acinetobacter calcoaceticus cultures.
Acinetobacter calcoaceticus BD413 produces variable amounts of an exocellular lipase that becomes rapidly inactivated upon secretion. To achieve high yield and protect the enzyme, we assayed the addition of several inert compounds to cell-free supernatants, cell fractions, and whole cultures. Glass beads, poly(ethylene glycol) 600, Triton X-100, saccharose, gum arabic, and beta-cyclodextrin were among the compounds tested. beta-Cyclodextrin and gum arabic (and saccharose to a lesser extent) were effective enzyme stabilizers in cell-free supernatants, while gum arabic, glass beads, and Triton X-100 improved lipase secretion from cells, and, therefore, total lipase yield (30-50%, according to the additive). In whole cultures, beta-cyclodextrin was the most effective additive, particularly in combination with glass beads or gum arabic. Indeed, cultures containing beta-cyclodextrin plus gum arabic were able to maintain 95% (+/- 1.5%) of the initial lipase activity for more than 16 h, while control cultures with no additives maintained only 10% (+/- 4%) of the enzyme activity after the same period. In conclusion, the addition of inert compounds in cultures may be considered a useful approach for achieving increased yield and lipase stabilization, amenable for downstream processing.
Martinez
Daniela A
DA
Cátedra de Microbiología Industrial y Biotecnología, Facultad de Farmacia y Bioquímica, Universidad de Buenos Aires, Junín 956 (1113) Buenos Aires, Argentina.
Nudel
B Clara
BC
D016954:Acinetobacter calcoaceticus / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development; D018533:Biomass; D003470:Culture Media; D003505:Cyclodextrins / Q000378:metabolism; D004795:Enzyme Stability; D005898:Glass / Q000737:chemistry; D006170:Gum Arabic / Q000378:metabolism; D007218:Industrial Microbiology; D008049:Lipase / Q000378:metabolism*; D017830:Octoxynol / Q000378:metabolism; D011092:Polyethylene Glycols / Q000378:metabolism; D013395:Sucrose / Q000378:metabolism; D013997:Time Factors; D047392:beta-Cyclodextrins; D001616:beta-Galactosidase / Q000032:analysis
D003470:Culture Media; D003505:Cyclodextrins; D047392:beta-Cyclodextrins; D011092:Polyethylene Glycols; D013395:Sucrose; D006170:Gum Arabic; D017830:Octoxynol; D008049:Lipase; D001616:beta-Galactosidase; C031215:betadex