Protease purification from Bacillus amyloliquefaciens B7 using aqueous two phase system
Bacillus amyloliquefaciens B7 was isolated from fish fermented sauce and identified as protease producer. Generally, in downstream processing, purification of enzyme consumes high cost in terms of reagents and equipment used. Moreover, harsh purification methods used might cause denaturation of en...
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| Main Authors: | , , , |
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| Format: | Conference or Workshop Item |
| Language: | English |
| Published: |
Kulliyah of Engineering, International Islamic University Malaysia
2016
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| Subjects: | |
| Online Access: | http://irep.iium.edu.my/51790/1/51790.pdf http://irep.iium.edu.my/51790/ http://www.iium.edu.my/icbioe/2016/ |
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| Summary: | Bacillus amyloliquefaciens B7 was isolated from fish fermented sauce and identified as protease producer.
Generally, in downstream processing, purification of enzyme consumes high cost in terms of reagents and
equipment used. Moreover, harsh purification methods used might cause denaturation of enzymes.
Therefore, there is a high demand for efficient and low-cost extraction and purification methods. Aqueous
two-phase system (ATPS) is an alternative that should be considered as it is simple, rapid separation yet
cause little denaturation. Protease produced by B. amyloliquefaciens B7 was partitioned in two different
ATPS, which are PEG/potassium phosphate and PEG/sodium citrate, and best separation was found in
PEG/sodium citrate based on protease specific activity. Protease activity was determined using casein as the
substrate. Using response surface methodology design, two independent variables which are pH and
temperature were varied during protease purification by aqueous two-phase system composed of PEG
1500/sodium citrate and the highest enzyme activity was found at the interface phase. Optimum purification
method observed is at pH 6, 30OC, 27 % (w/w) of PEG and 34 % (w/w) of Sodium Citrate and the highest
enzyme activity achieved is 0.19± 0.010 U/ml. |
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