Characteristics of recombinant maltogenic amylase from Geobacillus sp. SK70
A thermostable maltogenic amylase producing-bacterium was isolated from a 70°C hot spring at Sungai Klah, Perak, Malaysia and was designated as Geobacillus sp. SK70 based on the 16S rRNA gene analysis. The gene encoding a thermostable maltogenic amylase was expressed in Escherichia coli using pET1...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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National Institute of Science Communication and Information Resources
2017
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| Online Access: | http://psasir.upm.edu.my/id/eprint/61085/1/Characteristics%20of%20recombinant%20maltogenic%20amylase%20from%20Geobacillus%20sp.%20SK70.pdf |
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| author | Sulong, Moohamad Ropaning Leow, Adam Thean Chor Raja Abdul Rahman, Raja Noor Zaliha Basri, Mahiran Salleh, Abu Bakar |
| author_facet | Sulong, Moohamad Ropaning Leow, Adam Thean Chor Raja Abdul Rahman, Raja Noor Zaliha Basri, Mahiran Salleh, Abu Bakar |
| author_sort | Sulong, Moohamad Ropaning |
| collection | UPM |
| description | A thermostable maltogenic amylase producing-bacterium was isolated from a 70°C hot spring at Sungai Klah, Perak, Malaysia and was designated as Geobacillus sp. SK70 based on the 16S rRNA gene analysis. The gene encoding a thermostable maltogenic amylase was expressed in Escherichia coli using pET102 Directional TOPO expression vector, and it is the first ever report on using such expression vector. The highest enzyme activity was obtained after 12 h of post-induction time using 0.02 mM isopropyl β-D-thiogalactopyranoside (IPTG). The enzyme was purified to homogeneity with 8.2-fold and 41% recovery through a single-step using His-Trap HP affinity column chromatography. The optimum temperature and pH of the purified enzyme was at 55°C and pH 7.0, respectively, and showed broad pH stability ranging from pH 5.0 to 10.0. The activity of the purified enzyme was stable in the presence of 1 mM Ca2+;stimulated by 1 mM Mn2+and Zn2+, and 0.1 % (v/v) Tween-20; and inhibited by 1% (v/v) of 2-mercaptolethanol, EDTA and SDS. Thus the enzyme could be considered Ca2+-independent, which demonstrated characteristic unlike other reported maltogenic amylases, and offered good characteristics for industrial applications. |
| first_indexed | 2024-03-06T09:39:38Z |
| format | Article |
| id | upm.eprints-61085 |
| institution | Universiti Putra Malaysia |
| language | English |
| last_indexed | 2024-03-06T09:39:38Z |
| publishDate | 2017 |
| publisher | National Institute of Science Communication and Information Resources |
| record_format | dspace |
| spelling | upm.eprints-610852019-05-06T07:47:03Z http://psasir.upm.edu.my/id/eprint/61085/ Characteristics of recombinant maltogenic amylase from Geobacillus sp. SK70 Sulong, Moohamad Ropaning Leow, Adam Thean Chor Raja Abdul Rahman, Raja Noor Zaliha Basri, Mahiran Salleh, Abu Bakar A thermostable maltogenic amylase producing-bacterium was isolated from a 70°C hot spring at Sungai Klah, Perak, Malaysia and was designated as Geobacillus sp. SK70 based on the 16S rRNA gene analysis. The gene encoding a thermostable maltogenic amylase was expressed in Escherichia coli using pET102 Directional TOPO expression vector, and it is the first ever report on using such expression vector. The highest enzyme activity was obtained after 12 h of post-induction time using 0.02 mM isopropyl β-D-thiogalactopyranoside (IPTG). The enzyme was purified to homogeneity with 8.2-fold and 41% recovery through a single-step using His-Trap HP affinity column chromatography. The optimum temperature and pH of the purified enzyme was at 55°C and pH 7.0, respectively, and showed broad pH stability ranging from pH 5.0 to 10.0. The activity of the purified enzyme was stable in the presence of 1 mM Ca2+;stimulated by 1 mM Mn2+and Zn2+, and 0.1 % (v/v) Tween-20; and inhibited by 1% (v/v) of 2-mercaptolethanol, EDTA and SDS. Thus the enzyme could be considered Ca2+-independent, which demonstrated characteristic unlike other reported maltogenic amylases, and offered good characteristics for industrial applications. National Institute of Science Communication and Information Resources 2017 Article PeerReviewed text en http://psasir.upm.edu.my/id/eprint/61085/1/Characteristics%20of%20recombinant%20maltogenic%20amylase%20from%20Geobacillus%20sp.%20SK70.pdf Sulong, Moohamad Ropaning and Leow, Adam Thean Chor and Raja Abdul Rahman, Raja Noor Zaliha and Basri, Mahiran and Salleh, Abu Bakar (2017) Characteristics of recombinant maltogenic amylase from Geobacillus sp. SK70. Indian Journal of Biotechnology, 16 (1). 91 - 99. ISSN 0972-5849 http://nopr.niscair.res.in/bitstream/123456789/42299/1/IJBT%2016%281%29%2091-99.pdf |
| spellingShingle | Sulong, Moohamad Ropaning Leow, Adam Thean Chor Raja Abdul Rahman, Raja Noor Zaliha Basri, Mahiran Salleh, Abu Bakar Characteristics of recombinant maltogenic amylase from Geobacillus sp. SK70 |
| title | Characteristics of recombinant maltogenic amylase from Geobacillus sp. SK70 |
| title_full | Characteristics of recombinant maltogenic amylase from Geobacillus sp. SK70 |
| title_fullStr | Characteristics of recombinant maltogenic amylase from Geobacillus sp. SK70 |
| title_full_unstemmed | Characteristics of recombinant maltogenic amylase from Geobacillus sp. SK70 |
| title_short | Characteristics of recombinant maltogenic amylase from Geobacillus sp. SK70 |
| title_sort | characteristics of recombinant maltogenic amylase from geobacillus sp sk70 |
| url | http://psasir.upm.edu.my/id/eprint/61085/1/Characteristics%20of%20recombinant%20maltogenic%20amylase%20from%20Geobacillus%20sp.%20SK70.pdf |
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