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59670

PURIFICATION, CHARACTERIZATION AND SEQUENCING OF AMYLASE A FROM ALKALIPHLIC BACILLUS LICHENIFORMIS STRAIN-MK7

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Last updated: 22 Jan 2023

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Abstract

Two degenerative primers were designated from the closest Bacillus, Bacillus licheniformis strain DSM13 (ATCC 14580) which has complete genome to get amylase gene from our Bacillus licheniformis strain MK7 (KP322016) by PCR technique. We got about 1500 bp PCR, the sequence analysis revealed that the alkaliphilic cellulase belongs to glycosyl hydrolase family 12A which has 100 % identity to glycoside hydrolase from Bacillus sp. (WP009329478) and has 99 % identity to family 12 xyloglucanase from Bacillus licheniformis (2JEMIA). The nucleotide sequence of the alkaliphilic cellulase gene (cel12A) was deposited to the GenBank under accession number (KP322015). The cel12A gene from B. licheniformis strain MK7 has open reading frame of 1500 bp that codes for 449 amino acid which has a molecular weight of 55KDa protein. The enzyme protein was further purified using ammonium sulphate precipitation and chromatography. The characterization of amylase protein showed that the enzyme has an optimum temperature at (37oC), optimum pH (9.0) and Some metals increase the enzyme activity such as CoCl2 +, K+ and EDTA , but others decrease the activity such as Ag +, Zn2 + and urea ions.

DOI

10.21608/rjab.2016.59670

Keywords

Amylase A, Purification, Characterization, Bacillus licheniformis strain –MK7

Authors

First Name

Mohamed

Last Name

kasem

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-

Affiliation

Microbial biotechnology dept., Genetic Engineering and Biotechnology Research Institute, University of Sadat City (USC), Egypt

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Orcid

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First Name

Aysam

Last Name

Fayed

MiddleName

M.

Affiliation

Molecular biology dept., Genetic Engineering and Biotechnology Research Institute, University of Sadat City (USC), Egypt

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Orcid

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First Name

Samah

Last Name

Eldemery

MiddleName

-

Affiliation

Molecular biology dept., Genetic Engineering and Biotechnology Research Institute, University of Sadat City (USC), Egypt

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Orcid

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First Name

Adel

Last Name

Okba

MiddleName

-

Affiliation

Microbial biotechnology dept., Genetic Engineering and Biotechnology Research Institute, University of Sadat City (USC), Egypt

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Orcid

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First Name

Atef

Last Name

Ibrahim

MiddleName

-

Affiliation

MicrobialBiotechnology Dept., Genetic Engineering & Biotechnology Research Institute (GEBRI), University of Sadat City (USC), Sadat City, Egypt

Email

atef.ibrahim@gebri.usc.edu.eg

City

Sadat City

Orcid

-

Volume

2

Article Issue

Special issue (2) for the first International Conference of Genetic Engineering and Biotechnology

Related Issue

5730

Issue Date

2016-06-01

Receive Date

2019-11-17

Publish Date

2016-06-01

Page Start

11

Page End

20

Print ISSN

2356-9433

Online ISSN

2682-3470

Link

https://rjab.journals.ekb.eg/article_59670.html

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https://rjab.journals.ekb.eg/service?article_code=59670

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2

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Original Article

Type Code

890

Publication Type

Journal

Publication Title

Research Journal of Applied Biotechnology

Publication Link

https://rjab.journals.ekb.eg/

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Article

Created At

22 Jan 2023