Extraction, Partial Purification and Characterization of Amylase from Apple (Malus Pumila)

International Journal of Food and Nutritional Science, Vol. 5, No. 3, July 2016

8 Pages Posted: 21 Mar 2017

See all articles by Harnek Saini

Harnek Saini

Kurukshetra University

Ritu Saini

Chaudhary Charan Singh - Haryana Agricultural University (CCS HAU)

Anjali Dahiya

Chaudhary Charan Singh - Haryana Agricultural University (CCS HAU)

Shikha Mehta

Chaudhary Charan Singh - Haryana Agricultural University (CCS HAU)

Date Written: July 2, 2016

Abstract

Amylases are among the most important enzymes and are of great significance in present day biotechnology and the spectrum of applications has widened many other fields such as clinical, medical and analytical practices. Malus pumila (apple) was homogenized in buffer for α-amylase extraction.

The crude extract showed 0.2 U/mg specific activity that was when subjected to ammonium sulfate precipitation, 0.4 U/mg specific activity was obtained. After applied to sephadex G-100 for gel filtration chromatography, it indicated the specific activity of 4.76 U/mg with 24-fold purification. NATIVE-PAGE of enzyme showed that unwanted proteins are removed and clear band of enzyme was appeared. The enzyme was partially purified and had a high activity in a broad pH range of 6-8, with the maximal activity occurring at pH 6. The enzyme was stable between the temperatures of 30-50 ºC and the optimum temperature for maximal amylase activity was found to be 35 ºC, after which the enzyme activity dropped. In apple, amylase activity was increased by the addition of Ca2 and Mn2 and Mg2 while Hg2 showed inhibitory effect. The enzyme retained its 90% activity after incubation for 1 h at temperature of 40 ºC.

Keywords: Amylase; Apple; Malus pumila; Purification; Characterization

Suggested Citation

Saini, Harnek and Saini, Ritu and Dahiya, Anjali and Mehta, Shikha, Extraction, Partial Purification and Characterization of Amylase from Apple (Malus Pumila) (July 2, 2016). International Journal of Food and Nutritional Science, Vol. 5, No. 3, July 2016. Available at SSRN: https://ssrn.com/abstract=2935593

Harnek Saini

Kurukshetra University ( email )

Kurukshetra University
kurukshetra (Haryana), Haryana 136119
India

Ritu Saini

Chaudhary Charan Singh - Haryana Agricultural University (CCS HAU)

Hisar
Haryana, 125004
India

Anjali Dahiya (Contact Author)

Chaudhary Charan Singh - Haryana Agricultural University (CCS HAU) ( email )

Hisar
Haryana, 125004
India

Shikha Mehta

Chaudhary Charan Singh - Haryana Agricultural University (CCS HAU)

Hisar
Haryana, 125004
India

Register to save articles to
your library

Register

Paper statistics

Downloads
79
Abstract Views
322
rank
305,657
PlumX Metrics