title: Purification and Properties of Extracellular Lipases with Transesterification Activity and 1,3-Regioselectivity fromRhizomucor mieheiandRhizopus oryzae.
abstract: Rhizomucor mieheiNRRL 5282 andRhizopus oryzaeNRRL 1526 can produce lipases with high synthetic activities in wheat bran-based solid-state culture. In this study, the purification and biochemical characterization of the lipolytic activities of these lipases are presented. SDS-PAGE indicated a molecular mass of about 55 and 35 kDa for the purifiedR. mieheiandRh. oryzaeenzymes, respectively.p-Nitrophenyl palmitate (pNPP) hydrolysis was maximal at 40°C and pH 7.0 for theR. mieheilipase, and at 30°C and pH 5.2 for theRh. oryzaeenzyme. The enzymes showed almost equal affinity topNPP, but theVmaxof theRh. oryzaelipase was about 1.13 times higher than that determined forR. mieheiusing the same substrate. For both enzymes, a dramatic loss of activity was observed in the presence of 5 mM Hg2+, Zn2+, or Mn2+, 10 mMN-bromosuccinimide or sodium dodecyl sulfate, and 5-10% (v/v) of hexanol or butanol. At the same time, they proved to be extraordinarily stable in the presence ofn-hexane, cyclohexane,n-heptane, and isooctane. Moreover, isopentanol up to 10% (v/v) and propionic acid in 1 mM concentrations increased thepNPP hydrolyzing activity ofR. mieheilipase. Both enzymes had 1,3-regioselectivity, and efficiently hydrolyzedp-nitrophenyl (pNP) esters with C8-C16 acids, exhibiting maximum activity towardspNP-caprylate (R. miehei) and pNP-dodecanoate (Rh. oryzae). The purified lipases are promising candidates for various biotechnological applications.
authors: Department of Microbiology, Faculty of Science and Informatics, University of Szeged, H-6726 Szeged, Közép fasor 52, Hungary.
authors: Miklós
authors: M
authors: Takó
authors: Department of Microbiology, Faculty of Science and Informatics, University of Szeged, H-6726 Szeged, Közép fasor 52, Hungary.
authors: Alexandra
authors: A
authors: KotogÁn
authors: Department of Microbiology, Faculty of Science and Informatics, University of Szeged, H-6726 Szeged, Közép fasor 52, Hungary.|HAS-USZ "Momentum" Fungal Pathogenicity Mechanisms Research Group, University of Szeged, H-6726 Szeged, Közép fasor 52, Hungary.
authors: TamÁs
authors: T
authors: Papp
authors: Department of Botany and Microbiology, College of Science, King Saud University, Riyadh-11451, Saudi Arabia.
authors: Shine
authors: S
authors: Kadaikunnan
authors: Department of Botany and Microbiology, College of Science, King Saud University, Riyadh-11451, Saudi Arabia.
authors: Naiyf S
authors: NS
authors: Alharbi
authors: Department of Microbiology, Faculty of Science and Informatics, University of Szeged, H-6726 Szeged, Közép fasor 52, Hungary.
authors: Csaba
authors: C
authors: VÁgvölgyi
mesh_terms: D001974:Bromosuccinimide / Q000494:pharmacology; D000440:Butanols / Q000494:pharmacology; D002210:Caprylates / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D004951:Esterification; D006536:Heptanes / Q000494:pharmacology; D006586:Hexanes / Q000494:pharmacology; D000441:Hexanols / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D007700:Kinetics; D007848:Laurates / Q000494:pharmacology; D008049:Lipase / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D008345:Manganese / Q000494:pharmacology; D008628:Mercury / Q000494:pharmacology; D009578:Nitrobenzenes / Q000494:pharmacology; D010168:Palmitates / Q000378:metabolism; D011422:Propionates / Q000494:pharmacology; D020103:Rhizomucor / Q000201:enzymology* / Q000235:genetics; D012233:Rhizopus / Q000201:enzymology* / Q000235:genetics; D012967:Sodium Dodecyl Sulfate / Q000494:pharmacology; D015032:Zinc / Q000494:pharmacology
chemical_list: C484326:4-nitrophenyl dodecanoate; D000440:Butanols; D002210:Caprylates; D006536:Heptanes; D006586:Hexanes; D000441:Hexanols; D007848:Laurates; D009578:Nitrobenzenes; D010168:Palmitates; D011422:Propionates; C050015:4-nitrophenyl palmitate; C034106:4-nitrophenyloctanoate; C026385:n-hexane; D012967:Sodium Dodecyl Sulfate; D008345:Manganese; C028618:n-heptane; D008049:Lipase; D008628:Mercury; D015032:Zinc; C029658:propionic acid; D001974:Bromosuccinimide
keywords: Zygomycetes; enzyme purification; extracellular lipase; lipolysis characterization; synthetic activity