Cloning and Expression of Gene Encoding L-Asparaginase from Local AnsZ Bacillus subtilis  

Oleh:

Azara, Rima ; Helianti, Is ; Kusnadi, Joni ; Yunianta

Jenis: Article from Journal - ilmiah nasional - terakreditasi DIKTI
Dalam koleksi: Microbiology Indonesia vol. 08 no. 2 (Jun. 2014), page 41-47.
Topik: Bacillus subtilis; expression; cloning; L-asparaginase (ansZ)
Fulltext: 274-823-1-PB_Ros.pdf (803.41KB)

Isi artikel

L-asparaginase is an enzyme that catalyzes the hydrolysis of L-asparagine into L-aspartic acid and ammonia. In medical aspect, L-asparaginase especially those came from and used as chemotherapy agent of acute lymphoblastic leukemia (ALL). However, new potential organisms possessing Lasparaginase production capacity with a similar therapeutic effect are still required. In strain 168, there are two kinds of L-asparaginase gene, and . The study of the later L-asparaginase ( ) has not been conducted intensively. The aim of this study is, first, to isolate this gene of L-asparaginase ( ) from local and then to express this gene in . Using PCR-cloning method, an open reading frame (ORF) containing 1128 bp was obtained. The ORF has 99% homology with sequence of Lasparaginase from Bsn5. The gene then was subcloned into pET 21d (+) with his -tag in the Cterminal of the gene product and expressed in BL21. L-asparaginase activity analyses showed that recombinant containing recombinant plasmid with open reading frame (ORF) L-asparaginase ( ) from had higher activity than that is not containingORF L-asparaginase ( ). Purificationwith HisPur Ni-NTAPurification Kit increased the specific activity of recombinant L-asparaginase ( ) enzyme 29 fold.

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