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Purification and properties of midgut α-amylase isolated from Morimus funereus (Coleoptera: Cerambycidae) larvae
dc.creator | Dojnov, Biljana | |
dc.creator | Božić, Nataša | |
dc.creator | Nenadović, V. | |
dc.creator | Ivanović, J. | |
dc.creator | Vujčić, Zoran | |
dc.date.accessioned | 2019-01-30T17:18:41Z | |
dc.date.available | 2019-01-30T17:18:41Z | |
dc.date.issued | 2008 | |
dc.identifier.issn | 1096-4959 | |
dc.identifier.uri | https://cer.ihtm.bg.ac.rs/handle/123456789/459 | |
dc.description.abstract | Using soluble starch as a substrate five isoforms of α-amylase were identified in a crude extract of Morimus funereus larvae. The main α-amylase (termed AMF-3) was purified by gel filtration chromatography and anion exchange chromatography to obtain a single band on sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). Its enzymatic purity was confirmed by an in-gel activity assay after SDS-PAGE. The purity of AMF-3 was increased 112-fold with a 15.4% yield. AMF-3 had apparent molecular masses of 33 and 31 kDa when analysed using SDS-PAGE and Superdex 75 FPLC gel filtration chromatography, respectively and a calculated isoelectric point of 3.2. Purified AMF-3 showed maximal activity at pH 5.2 and had an optimum activity temperature of 45 °C. AMF-3 retained over 90% of its maximum activity at temperatures from 45 to 60 °C. AMF-3 exhibited a high affinity towards soluble starch with a Km value of 0.43 mg/mL. Maximal AMF-3 activity was achieved in the presence of 0.1 mM CaCl2, while at higher concentrations its activity decreased. AMF-3 activity increased with increasing NaCl concentration. AMF-3 activity was significantly inhibited by α-amylase wheat inhibitor. Using a number of raw starch substrates maximum AMF-3 activity was achieved with horse-radish starch, in contrast to undetectable activity towards potato starch. | en |
dc.publisher | Elsevier Science Inc, New York | |
dc.relation | info:eu-repo/grantAgreement/MESTD/MPN2006-2010/142026/RS// | |
dc.rights | restrictedAccess | |
dc.source | Comparative Biochemistry and Physiology - B Biochemistry and Molecular Biology | |
dc.subject | Cerambycid beetle | en |
dc.subject | Isoform | en |
dc.subject | Midgut a-amylase | en |
dc.subject | Morimus funereus | en |
dc.subject | Xylophagous larvae | en |
dc.title | Purification and properties of midgut α-amylase isolated from Morimus funereus (Coleoptera: Cerambycidae) larvae | en |
dc.type | article | |
dc.rights.license | ARR | |
dcterms.abstract | Ненадовић, В.; Божић, Наташа; Дојнов, Биљана; Ивановић, Ј.; Вујчић, З.; | |
dc.citation.volume | 149 | |
dc.citation.issue | 1 | |
dc.citation.spage | 153 | |
dc.citation.epage | 160 | |
dc.citation.other | 149(1): 153-160 | |
dc.citation.rank | M22 | |
dc.identifier.pmid | 17942357 | |
dc.identifier.doi | 10.1016/j.cbpb.2007.09.009 | |
dc.identifier.scopus | 2-s2.0-36849076255 | |
dc.identifier.wos | 000254219600016 | |
dc.type.version | publishedVersion |