Abstract
Amylases are important industrial enzymes that have been applied widely in the food, detergent, and pulp industries and fermentation processes. In the present study, a gene encoding an alpha-amylase from the genomic DNA library of Paenibacillus sp. was identified and characterized. The amylase gene designated amy1 was shown to consist of 1,980 bp and shared sequence identity towards α-amylase genes from other Bacillus sp. The deduced amino acid sequence for Amy1 indicated 80 % sequence identity with other Bacillus strains. Heterologous expression of recombinant Amy1 in Escherichia coli BL21(DE3) facilitated the recovery of this protein in soluble form. Enzyme kinetic data revealed Amy1 to have a K m of 23.83 mg/mL and K cat of 48.74 min−1 and K cat /K m of 2 min−1 mg−1 mL−1 for starch. The activity of this protein was found to be enhanced by Mn2+, and furthermore, Amy1 remained active at a broad pH range (4–10) and temperature (30–90 °C). The ability of Amy1 to act on food waste under broad temperature and pH conditions, together with its ability to produce simple sugars, shows many advantages for further application in the food industry.
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Acknowledgments
This work at Konkuk University was partially supported by a National Research Foundation of Korea grant funded by the Korean Government (NRF-2011-619-E0002). This subject is also partially supported by the Korea Ministry of Environment as a “Converging Technology Project (201-101-007)” and as an “Eco-Innovation Project (405-112-0382).” This research was also supported by the 2012 KU Brain Pool of Konkuk University.
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Thangamani Rajesh and Yong Hyun Kim contributed equally to this work.
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Rajesh, T., Kim, Y.H., Choi, YK. et al. Identification and Functional Characterization of an α-Amylase with Broad Temperature and pH Stability from Paenibacillus sp.. Appl Biochem Biotechnol 170, 359–369 (2013). https://doi.org/10.1007/s12010-013-0197-z
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DOI: https://doi.org/10.1007/s12010-013-0197-z