Purification and properties of midgut α-amylase isolated from Morimus funereus (Coleoptera: Cerambycidae) larvae
Само за регистроване кориснике
2008
Чланак у часопису (Објављена верзија)
Метаподаци
Приказ свих података о документуАпстракт
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 C...aCl2, 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.
Кључне речи:
Cerambycid beetle / Isoform / Midgut a-amylase / Morimus funereus / Xylophagous larvaeИзвор:
Comparative Biochemistry and Physiology - B Biochemistry and Molecular Biology, 2008, 149, 1, 153-160Издавач:
- Elsevier Science Inc, New York
Финансирање / пројекти:
- Интеракције природних производа и њихових аналога са протеинима и нуклеинским киселинама (RS-142026)
DOI: 10.1016/j.cbpb.2007.09.009
ISSN: 1096-4959
PubMed: 17942357
WoS: 000254219600016
Scopus: 2-s2.0-36849076255
Институција/група
IHTMTY - JOUR AU - Dojnov, Biljana AU - Božić, Nataša AU - Nenadović, V. AU - Ivanović, J. AU - Vujčić, Zoran PY - 2008 UR - https://cer.ihtm.bg.ac.rs/handle/123456789/459 AB - 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. PB - Elsevier Science Inc, New York T2 - Comparative Biochemistry and Physiology - B Biochemistry and Molecular Biology T1 - Purification and properties of midgut α-amylase isolated from Morimus funereus (Coleoptera: Cerambycidae) larvae VL - 149 IS - 1 SP - 153 EP - 160 DO - 10.1016/j.cbpb.2007.09.009 ER -
@article{ author = "Dojnov, Biljana and Božić, Nataša and Nenadović, V. and Ivanović, J. and Vujčić, Zoran", year = "2008", 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.", publisher = "Elsevier Science Inc, New York", journal = "Comparative Biochemistry and Physiology - B Biochemistry and Molecular Biology", title = "Purification and properties of midgut α-amylase isolated from Morimus funereus (Coleoptera: Cerambycidae) larvae", volume = "149", number = "1", pages = "153-160", doi = "10.1016/j.cbpb.2007.09.009" }
Dojnov, B., Božić, N., Nenadović, V., Ivanović, J.,& Vujčić, Z.. (2008). Purification and properties of midgut α-amylase isolated from Morimus funereus (Coleoptera: Cerambycidae) larvae. in Comparative Biochemistry and Physiology - B Biochemistry and Molecular Biology Elsevier Science Inc, New York., 149(1), 153-160. https://doi.org/10.1016/j.cbpb.2007.09.009
Dojnov B, Božić N, Nenadović V, Ivanović J, Vujčić Z. Purification and properties of midgut α-amylase isolated from Morimus funereus (Coleoptera: Cerambycidae) larvae. in Comparative Biochemistry and Physiology - B Biochemistry and Molecular Biology. 2008;149(1):153-160. doi:10.1016/j.cbpb.2007.09.009 .
Dojnov, Biljana, Božić, Nataša, Nenadović, V., Ivanović, J., Vujčić, Zoran, "Purification and properties of midgut α-amylase isolated from Morimus funereus (Coleoptera: Cerambycidae) larvae" in Comparative Biochemistry and Physiology - B Biochemistry and Molecular Biology, 149, no. 1 (2008):153-160, https://doi.org/10.1016/j.cbpb.2007.09.009 . .