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Characteristics of recombinant maltogenic amylase from Geobacillus sp. SK70


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


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.

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Additional Metadata

Item Type: Article
Divisions: Faculty of Biotechnology and Biomolecular Sciences
Faculty of Science
Institute of Bioscience
Publisher: National Institute of Science Communication and Information Resources
Keywords: Ca2+-independent; Geobacillus sp.; Maltogenic amylase; thermostable; Zn activated
Depositing User: Ms. Nida Hidayati Ghazali
Date Deposited: 06 May 2019 07:47
Last Modified: 06 May 2019 07:47
URI: http://psasir.upm.edu.my/id/eprint/61085
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