CLONING AND EXPRESSION OF PENICILLIN ACYLASE FROM ARTHROBACTER VISCOSUS IN E. COLI

Q: How can I download an article?

To download an article from SID, first log in to the site, search for the article title, and click on the 'Download Article' option.

Q: How can I download an ISI article?

To download an ISI article on SID, enter the keyword or article title in the search bar, view the relevant results, click on the desired article, and select the 'Download Article' option.

Q: How can I access the SID database?

To access the SID database, visit SID.ir, create an account, and log in to access scientific resources.

Q: Is downloading articles from SID free?

Some articles on SID are available for free, while others require payment. Details are specified on the article's page.

EBRAHIMIPOUR GH.H.; BAMBAEI B.; KHODABANDEH M.; TORKINEZHAD F.

مرکز اطلاعات علمی Scientific Information Database (SID) - Trusted Source for Research and Academic Resources

Journal Paper

Paper Information

Journal: IRANIAN JOURNAL OF BIOLOGY Year:2009 | Volume:22 | Issue:1 Page(s): 1-11

Download Full-Text

Persian Verion

View:

2,933

Download:

Cites:

Information Journal Paper

Title

CLONING AND EXPRESSION OF PENICILLIN ACYLASE FROM ARTHROBACTER VISCOSUS IN E. COLI

Author(s)

EBRAHIMIPOUR GH.H. | BAMBAEI B. | KHODABANDEH M. | TORKINEZHAD F. | Issue Writer Certificate

Keywords

ARTHROBACTER VISCOSUSQ3

PENICILLIN G ACYLASEQ3

RECOMBINANT EXPRESSIONQ3

Abstract

Penicillin acylase (penicillin amidase, PAC; EC 3.5.1.11) is the key Enzyme used in the industrial production of b-lactam antibiotics. This enzyme cleaves Penicillin G to 6- amino penicillanic acid (6-APA) and PAA by hydrolysis of acyle side chain. It can process this reaction in both directions (hydrolysis and synthesis). Here, we sought to clone and express the PENICILLIN G ACYLASE gene from A. viscosus in Escherichia coli, to improve expression level. Following PCR amplification of PAC gene from genome of A. viscosus, PAC gene was cloned in pET26(b) expression vector in frame with DNA sequence coding for 6xHis tag presented at 3' of MCS of the vector. After sequencing and confirmation of gene identity with published sequence (L04471), positive clones were screened based on differential response of Serratia marcescens (ATCC27177) to penicillin G and 6-APA (resistance to penicillin G and susceptibility to 6-APA). Furthermore, SDS-PAGE and western blotting with anti.His.HRP. Conjugate antibodies confirmed the autocatalytic processing in cytoplasm to produce the active enzyme. Our results shows the ability of E.coli (BL21) strain in expression of active pac gene from a gram positive bacterium in the cytoplasm.. Furthermore, our results show the benefit of T7 promoter and IPTG induction in obtaining high expression levels in E.coli for PENICILLIN G ACYLASE from a gram positive bacterium.

Cites

No record.

References

No record.

Cite

APA: Copy

EBRAHIMIPOUR, GH.H., BAMBAEI, B., KHODABANDEH, M., & TORKINEZHAD, F.. (2009). CLONING AND EXPRESSION OF PENICILLIN ACYLASE FROM ARTHROBACTER VISCOSUS IN E. COLI. IRANIAN JOURNAL OF BIOLOGY, 22(1), 1-11. SID. https://sid.ir/paper/21425/en

Vancouver: Copy

EBRAHIMIPOUR GH.H., BAMBAEI B., KHODABANDEH M., TORKINEZHAD F.. CLONING AND EXPRESSION OF PENICILLIN ACYLASE FROM ARTHROBACTER VISCOSUS IN E. COLI. IRANIAN JOURNAL OF BIOLOGY[Internet]. 2009;22(1):1-11. Available from: https://sid.ir/paper/21425/en

IEEE: Copy

GH.H. EBRAHIMIPOUR, B. BAMBAEI, M. KHODABANDEH, and F. TORKINEZHAD, “CLONING AND EXPRESSION OF PENICILLIN ACYLASE FROM ARTHROBACTER VISCOSUS IN E. COLI,” IRANIAN JOURNAL OF BIOLOGY, vol. 22, no. 1, pp. 1–11, 2009, [Online]. Available: https://sid.ir/paper/21425/en

Related Journal Papers