The biochemical properties of the alkaline phosphatases (AlPs) produced by Rhizopus microsporus are described. High enzymic levels were produced within 1-2 d in agitated cultures with 1 % wheat bran. Intra- and extracellular AlPs were purified 5.0 and 9.3x, respectively, by DEAE-cellulose and ConA-sepharose chromatography. Molar mass of 118 and 120 kDa was estimated by gel filtration for both forms of phosphatases. SDS-PAGE indicated dimeric structures of 57 kDa for both forms. Mn(2+), Na(+) and Mg(2+) stimulated the activity, while Al(3+) and Zn(2+) activated only the extracellular form. Optimum temperature and pH for both phosphatases were 65 degrees C and pH 8.0, respectively. The enzymes were stable at 50 degrees C for at least 15 min. Hydrolysis of 4-nitrophenyl phosphate exhibited a K (m) 0.28 and 0.22 mmol/L, with upsilon (lim) 5.89 and 4.84 U/mg, for intra- and extracellular phosphatases, respectively. The properties of the reported AlPs may be suitable for biotechnological application.

• MeSH
• alkalická fosfatasa izolace a purifikace   metabolismus   MeSH
• chromatografie agarózová MeSH
• chromatografie DEAE-celulózová MeSH
• elektroforéza v polyakrylamidovém gelu MeSH
• fungální proteiny izolace a purifikace   metabolismus   MeSH
• gelová chromatografie MeSH
• kationty farmakologie   MeSH
• koncentrace vodíkových iontů MeSH
• molekulová hmotnost MeSH
• Rhizopus enzymologie   MeSH
• vysoká teplota MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• alkalická fosfatasa MeSH
• fungální proteiny MeSH
• kationty MeSH