Cloning and characterization of two new thermostable and alkalitolerant α-amylase from the Anoxybacillus species that produce high levels of maltose
Two genes that encode α-amylases from two Anoxybacillus species were cloned and expressed in Escherichia coli. The genes are 1,518 bp long and encode 506 amino acids. Both sequences are 98% similar but are distinct from other well-known α-amylases. Both of the recombinant enzymes, ASKA and ADTA, wer...
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| Format: | Article |
| Language: | English |
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2012
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| Online Access: | http://eprints.utm.my/46693/1/YenYenChai_2012_Cloning%20and%20characterization%20of%20two%20new%20thermostable.pdf |
| _version_ | 1796859004572401664 |
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| author | Chai, Yen Yen Raja Abd. Rahman, Raja Noor Zaliha Md. Illias, Rosli Goh, Kian Mau |
| author_facet | Chai, Yen Yen Raja Abd. Rahman, Raja Noor Zaliha Md. Illias, Rosli Goh, Kian Mau |
| author_sort | Chai, Yen Yen |
| collection | ePrints |
| description | Two genes that encode α-amylases from two Anoxybacillus species were cloned and expressed in Escherichia coli. The genes are 1,518 bp long and encode 506 amino acids. Both sequences are 98% similar but are distinct from other well-known α-amylases. Both of the recombinant enzymes, ASKA and ADTA, were purified using an α-CD–Sepharose column. They exhibited an optimum activity at 60°C and pH 8. Both amylases were stable at pH 6–10. At 60°C in the absence of Ca2+, negligible reduction in activity for up to 48 h was observed. The activity half-life at 65°C was 48 and 3 h for ASKA and ADTA, respectively. In the presence of Ca2+ ions, both amylases were highly stable for at least 48 h and had less than a 10% decrease in activity at 70°C. Both enzymes exhibited similar end-product profiles, and the predominant yield was maltose (69%) from starch hydrolysis. To the best of our knowledge, most α-amylases that produce high levels of maltose are active at an acidic to neutral pH. This is the first report of two thermostable, alkalitolerant recombinant α-amylases from Anoxybacillus that produce high levels of maltose and have an atypical protein sequence compared with known α-amylases. |
| first_indexed | 2024-03-05T19:20:46Z |
| format | Article |
| id | utm.eprints-46693 |
| institution | Universiti Teknologi Malaysia - ePrints |
| language | English |
| last_indexed | 2024-03-05T19:20:46Z |
| publishDate | 2012 |
| record_format | dspace |
| spelling | utm.eprints-466932017-09-18T04:00:31Z http://eprints.utm.my/46693/ Cloning and characterization of two new thermostable and alkalitolerant α-amylase from the Anoxybacillus species that produce high levels of maltose Chai, Yen Yen Raja Abd. Rahman, Raja Noor Zaliha Md. Illias, Rosli Goh, Kian Mau QR Microbiology Two genes that encode α-amylases from two Anoxybacillus species were cloned and expressed in Escherichia coli. The genes are 1,518 bp long and encode 506 amino acids. Both sequences are 98% similar but are distinct from other well-known α-amylases. Both of the recombinant enzymes, ASKA and ADTA, were purified using an α-CD–Sepharose column. They exhibited an optimum activity at 60°C and pH 8. Both amylases were stable at pH 6–10. At 60°C in the absence of Ca2+, negligible reduction in activity for up to 48 h was observed. The activity half-life at 65°C was 48 and 3 h for ASKA and ADTA, respectively. In the presence of Ca2+ ions, both amylases were highly stable for at least 48 h and had less than a 10% decrease in activity at 70°C. Both enzymes exhibited similar end-product profiles, and the predominant yield was maltose (69%) from starch hydrolysis. To the best of our knowledge, most α-amylases that produce high levels of maltose are active at an acidic to neutral pH. This is the first report of two thermostable, alkalitolerant recombinant α-amylases from Anoxybacillus that produce high levels of maltose and have an atypical protein sequence compared with known α-amylases. 2012 Article PeerReviewed application/pdf en http://eprints.utm.my/46693/1/YenYenChai_2012_Cloning%20and%20characterization%20of%20two%20new%20thermostable.pdf Chai, Yen Yen and Raja Abd. Rahman, Raja Noor Zaliha and Md. Illias, Rosli and Goh, Kian Mau (2012) Cloning and characterization of two new thermostable and alkalitolerant α-amylase from the Anoxybacillus species that produce high levels of maltose. Journal of Industrial Microbiology & Biotechnology, 39 . pp. 731-741. ISSN 1367-5435 http://dx.doi.org/10.1007/s10295-011-1074-9 https://doi.org/10.1007/s10295-011-1074-9 |
| spellingShingle | QR Microbiology Chai, Yen Yen Raja Abd. Rahman, Raja Noor Zaliha Md. Illias, Rosli Goh, Kian Mau Cloning and characterization of two new thermostable and alkalitolerant α-amylase from the Anoxybacillus species that produce high levels of maltose |
| title | Cloning and characterization of two new thermostable and alkalitolerant α-amylase from the Anoxybacillus species that produce high levels of maltose |
| title_full | Cloning and characterization of two new thermostable and alkalitolerant α-amylase from the Anoxybacillus species that produce high levels of maltose |
| title_fullStr | Cloning and characterization of two new thermostable and alkalitolerant α-amylase from the Anoxybacillus species that produce high levels of maltose |
| title_full_unstemmed | Cloning and characterization of two new thermostable and alkalitolerant α-amylase from the Anoxybacillus species that produce high levels of maltose |
| title_short | Cloning and characterization of two new thermostable and alkalitolerant α-amylase from the Anoxybacillus species that produce high levels of maltose |
| title_sort | cloning and characterization of two new thermostable and alkalitolerant α amylase from the anoxybacillus species that produce high levels of maltose |
| topic | QR Microbiology |
| url | http://eprints.utm.my/46693/1/YenYenChai_2012_Cloning%20and%20characterization%20of%20two%20new%20thermostable.pdf |
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