Molecular cloning, characterization, and homologous expression of an endochitinase gene from Bacillus thuringiensis serovar morrisoni

Okay, Sezer
Özcengiz, Gülay
The endochitinase gene (chi3023) of Bacillus thuringiensis (Bt) serovar morrisoni strain 3023 was amplified via polymerase chain reaction (PCR) and cloned in Escherichia coli. The ORF of chi3023 (GenBank Accession Number: DQ993175) consists of 2031 nucleotides encoding a 676-residue protein with a calculated molecular mass of 74.5 kDa and a pI value of 6.0. The amino acid sequence of Chi3023 was compared with previously sequenced Bt chitinases and the phylogenetic relationships among them were determined. The deduced N-terminal 34 amino acids of the premature Chi3023 exhibited a typical signal peptide. The E. coli-Bt shuttle vector pHT315 was used for homologous expression of chi3023. Introduction of recombinant pHT315BTC, carrying chi3023 into Bt serovar morrisoni 3023, resulted in a 23-fold increase in endochitinase activity (0.185 U/mg versus 4.256 U/mg).


Cloning and expression of gerE and glmS genes in Escherichia coli and Bacillus subtilis, and investigation of their possible interrelation with bacilysin biosynthesis
Akaysoy, Sergen; Özcengiz, Gülay; Department of Biology (2022-9-15)
Bacillus subtilis is the Gram-positive model bacterium that enzymatically produces the dipeptide antibiotic bacilysin. Bacilysin is the simplest bioactive peptide known composed of L-alanine at its N-terminal and L-anticapsin at its C-terminal. In a former work in our laboratory, a mutant strain of B. subtilis, namely OGU1 was constructed by bacA-targeted pMutin T3 insertion into the parental strain PY79 genome resulting in a genomic organization bacA′::lacZ::erm::bacABCDEF and unable to synthesize bacilysi...
Differential expressions and functions of phosphodiesterase enzymes in different regions of the rat heart
DERİCİ, MEHMET KÜRŞAT; SADİ, GÖKHAN; Cenik, Basar; Güray, Nülüfer Tülün; DEMİREL YILMAZ, EMİNE (Elsevier BV, 2019-02-05)
Phosphodiesterase enzymes (PDEs) are responsible for the adjustment of cyclic nucleotide levels. Alterations in PDE expressions in different tissues cause conflicts between functional and clinical effects of PDE inhibitors. Therefore, the aim of this study was to investigate the gene and protein expressions and the functional role of PDEs in atrium and ventricle of rat heart The expressions of PDEs were examined in cardiac intact tissues and enzymatically isolated cells. The effects of PDE1-5 inhibitors (vi...
Cloning, characterization and heterologous expression of the aspartokinase and aspartate semialdehyde dehydrogenase genes from the cephamycin C producer streptomyces clavuligerus
Tunca, Sedef; Özcengiz, Gülay; Piret, Jacqueline; Department of Biotechnology (2002)
The carbon flow through the lysine branch of aspartate pathway is a rate limiting step in the formation of cephamycin C, a broad spectrum P-lactam antibiotic produced by Streptomyces clavuligerus. In this study, the ask m(aspartokinase) and asd (aspartate semialdehyde dehydrogenase) genes which encode the enzymes catalyzing the first two steps of aspartate pathway were cloned and characterized as the first time from S. clavuligerus NRRL 3585. This was accomplished by constructing four mini-libraries of S. c...
Comparative analysis of product and by-product distributions in defined and complex media in serine alkaline protease production by recombinant Basillus subtilis
Oktar, Ceren; Çalık, Pınar; Department of Chemical Engineering (2003)
In this study, firstly the effects of aspartic acid group amino acids -which were reported to be the potential bottleneck in serine alkaline protease (SAP) synthesis- on SAP production were investigated by substituting at a concentration range of 0-15 mM by using recombinant Bacillus subtilis carrying pHV1434::subC gene. All aspartic acid group amino acids except threonine inhibited SAP activity when CAA= 2.5 mM. The highest SAP activities with asparagine, aspartic acid, lysine, threonine, isoleucine and me...
Molecular cloning, characterization, and expression analysis of a gene encoding a Ran binding protein (RanBP) in Cucumis melo L.
Baloglu, Mehmet Cengiz; Zakharov, Florence Negre; Öktem, Hüseyin Avni; Yücel, Ayşe Meral (2011-01-01)
Ran binding proteins (RanBPs) are highly conserved members of the GTP-binding protein family that are involved in nuclear protein export between the nucleus and the cytoplasm. In this study, a CmRanBP gene from a melon was isolated (Cucumis melo L.) using the RACE (rapid amplification of cDNA ends) method. The 778 basepair long melon, with a RanBP cDNA encoding consisting of 197 amino acids (22.2 kDa protein), was characterized (GenBank accession no: EU853459). The predicted amino acid sequence of CmRanBP w...
Citation Formats
S. Okay and G. Özcengiz, “Molecular cloning, characterization, and homologous expression of an endochitinase gene from Bacillus thuringiensis serovar morrisoni,” TURKISH JOURNAL OF BIOLOGY, pp. 1–7, 2011, Accessed: 00, 2020. [Online]. Available: