Cover Image

Extraction, purification and characterization of amylase enzyme from white pitaya (Hylocereus undatus (Haworth) britton & rose) peel using aqueous two- phase system

Amylase is one of the most important enzymes largely used in biotechnological and industrial applications. Thirty percent of the World’s enzyme production is accounted by Amylase. Malaysia has around 927.4 ha (363.2 ha production areas) pitaya fruit-growing areas produces about 2,534.2 tons worth ar...

Full description

Saved in:
Bibliographic Details
Main Author: Shad, Zahra
Format: Thesis
Language:English
Published: 2015
Online Access:http://psasir.upm.edu.my/id/eprint/64521/1/FSTM%202015%2023%20edited.pdf
http://psasir.upm.edu.my/id/eprint/64521/
Tags: Add Tag
No Tags, Be the first to tag this record!
id my.upm.eprints.64521
record_format eprints
spelling my.upm.eprints.645212018-11-08T07:43:10Z http://psasir.upm.edu.my/id/eprint/64521/ Extraction, purification and characterization of amylase enzyme from white pitaya (Hylocereus undatus (Haworth) britton & rose) peel using aqueous two- phase system Shad, Zahra Amylase is one of the most important enzymes largely used in biotechnological and industrial applications. Thirty percent of the World’s enzyme production is accounted by Amylase. Malaysia has around 927.4 ha (363.2 ha production areas) pitaya fruit-growing areas produces about 2,534.2 tons worth around US$3.5 million. Peels are one of the byproducts that are obtained from the processing of pitaya. Pitaya peel is mostly a waste material from fruit and beverage industries. Although it consists about 33% of whole fruit by weight and possesses valuable enzymes such as amylase, it is not being presently used commercially and is considered as a waste. It could have been efficiently used for commercial and economical production of natural enzymes. Therefore, this research studied the extraction, purification and characterization of amylase from white pitaya (Hylocereusundatus) peel. Extraction of amylase from peel of white pitaya was optimized using full factorial design (FFD) with three variables, sodium phosphate buffer (pH 4.5-7.5), mixing time (1- 3min) and buffer to sample ratio (1:3-1:5). The purification was carried out using aqueous two phase system (ATPS). The effectiveness of different parameters on purification and selective separation, such as polyethylene glycol (PEG) molecular weight (4000 to 8000), PEG concentration (10 to 18%), sodium citrate concentration (12-20%) and NaCl (2-8%) were optimized using response surface methodology (RSM). The purified amylase enzyme was characterized based on pH, temperature and metal ions, surfactants and oxidizing agents. It was found that optimum condition for amylase extraction was with sodium phosphate buffer pH 6 and buffer to sample ratio of (1:4) for 2 min which yielded the enzyme with specific activity of 5.89 U/mg. The purification of amylase was studied with ATPS method polyethylene glycol/sodium citrate. The optimum purification factor and yield were obtained when 14% (w/w) of polyethylene glycol 6000 g/mol, 16 % (w/w) sodium citrate buffer and 5% of NaCl were used. Amylase purification factor and yield using ATPS were 4.43 and 89.12%, respectively. The optimum temperature and pH activity of amylase were 55 °C and 6, respectively. This enzyme was also stable in the presence of surfactants and oxidizing agents. Moreover, it was found that the activity of amylase was increased in the presence of calcium ions. The unique characteristics of amylase from white pitaya peel indicate the great potential application of the enzyme in food and biotechnology industries. 2015-11 Thesis NonPeerReviewed text en http://psasir.upm.edu.my/id/eprint/64521/1/FSTM%202015%2023%20edited.pdf Shad, Zahra (2015) Extraction, purification and characterization of amylase enzyme from white pitaya (Hylocereus undatus (Haworth) britton & rose) peel using aqueous two- phase system. Masters thesis, Universiti Putra Malaysia.
institution Universiti Putra Malaysia
building UPM Library
collection Institutional Repository
continent Asia
country Malaysia
content_provider Universiti Putra Malaysia
content_source UPM Institutional Repository
url_provider http://psasir.upm.edu.my/
language English
description Amylase is one of the most important enzymes largely used in biotechnological and industrial applications. Thirty percent of the World’s enzyme production is accounted by Amylase. Malaysia has around 927.4 ha (363.2 ha production areas) pitaya fruit-growing areas produces about 2,534.2 tons worth around US$3.5 million. Peels are one of the byproducts that are obtained from the processing of pitaya. Pitaya peel is mostly a waste material from fruit and beverage industries. Although it consists about 33% of whole fruit by weight and possesses valuable enzymes such as amylase, it is not being presently used commercially and is considered as a waste. It could have been efficiently used for commercial and economical production of natural enzymes. Therefore, this research studied the extraction, purification and characterization of amylase from white pitaya (Hylocereusundatus) peel. Extraction of amylase from peel of white pitaya was optimized using full factorial design (FFD) with three variables, sodium phosphate buffer (pH 4.5-7.5), mixing time (1- 3min) and buffer to sample ratio (1:3-1:5). The purification was carried out using aqueous two phase system (ATPS). The effectiveness of different parameters on purification and selective separation, such as polyethylene glycol (PEG) molecular weight (4000 to 8000), PEG concentration (10 to 18%), sodium citrate concentration (12-20%) and NaCl (2-8%) were optimized using response surface methodology (RSM). The purified amylase enzyme was characterized based on pH, temperature and metal ions, surfactants and oxidizing agents. It was found that optimum condition for amylase extraction was with sodium phosphate buffer pH 6 and buffer to sample ratio of (1:4) for 2 min which yielded the enzyme with specific activity of 5.89 U/mg. The purification of amylase was studied with ATPS method polyethylene glycol/sodium citrate. The optimum purification factor and yield were obtained when 14% (w/w) of polyethylene glycol 6000 g/mol, 16 % (w/w) sodium citrate buffer and 5% of NaCl were used. Amylase purification factor and yield using ATPS were 4.43 and 89.12%, respectively. The optimum temperature and pH activity of amylase were 55 °C and 6, respectively. This enzyme was also stable in the presence of surfactants and oxidizing agents. Moreover, it was found that the activity of amylase was increased in the presence of calcium ions. The unique characteristics of amylase from white pitaya peel indicate the great potential application of the enzyme in food and biotechnology industries.
format Thesis
author Shad, Zahra
spellingShingle Shad, Zahra
Extraction, purification and characterization of amylase enzyme from white pitaya (Hylocereus undatus (Haworth) britton & rose) peel using aqueous two- phase system
author_facet Shad, Zahra
author_sort Shad, Zahra
title Extraction, purification and characterization of amylase enzyme from white pitaya (Hylocereus undatus (Haworth) britton & rose) peel using aqueous two- phase system
title_short Extraction, purification and characterization of amylase enzyme from white pitaya (Hylocereus undatus (Haworth) britton & rose) peel using aqueous two- phase system
title_full Extraction, purification and characterization of amylase enzyme from white pitaya (Hylocereus undatus (Haworth) britton & rose) peel using aqueous two- phase system
title_fullStr Extraction, purification and characterization of amylase enzyme from white pitaya (Hylocereus undatus (Haworth) britton & rose) peel using aqueous two- phase system
title_full_unstemmed Extraction, purification and characterization of amylase enzyme from white pitaya (Hylocereus undatus (Haworth) britton & rose) peel using aqueous two- phase system
title_sort extraction, purification and characterization of amylase enzyme from white pitaya (hylocereus undatus (haworth) britton & rose) peel using aqueous two- phase system
publishDate 2015
url http://psasir.upm.edu.my/id/eprint/64521/1/FSTM%202015%2023%20edited.pdf
http://psasir.upm.edu.my/id/eprint/64521/
_version_ 1643838048515915776
score 13.211869

Similar Items