Bacillus subtilis overproduces industrially important extracellular enzymes upon the targeted deletion of bacilysin biosynthetic operon

Özcengiz, Gülay
Islerel, E. Tekin
Aktas, C.
Bacilysin being produced by Bacillus subtilis is the smallest peptide antibiotic ever known. It is composed of an N-terminal l-alanine and a modified amino acid at its C-terminal, namely anticapsin. bacABCDEF operon and a monocistronic gene bacG are functional for bacilysin production in the organism, bacABCDFG being needed for the flux from prephenate to anticapsin and then to mature bacilysin while bacE gene within the operon is involved in resistance of the producer by pumping bacilysin out of the cell. Our earlier studies demonstrated that quorum sensing global regulation operates in bacilysin biosynthesis through the action of ComQ/ComX, PhrC (CSF), ComP/ComA and molecular regulation also requires an intact surfactin biosynthetic operon, srfA. We recently performed a dynamic secretome analysis of B. subtilis PY79 and its bac operon-deleted derivative OGU1 by taking 2DE MALDI TOF/MS and LC–MS MS as complementary approaches and identified ca. 200 proteins (extracellular, membrane and wall-associated proteins) differentially expressed between two strains. Since B. subtilis is one of the most important cell-factories with a significant capacity to produce a wide range of extracellular enzymes, of biotechnological interest was a significant increment in levels of the industrially-important extracellular enzymes upon the deletion of bac operon. These enzymes included chitosanase, arabinanase, levanase, lipase, phytase, endonuclease, bacillopeptidase F and minor extracellular protease. In this report, the results of quantitative transcript analysis of the respective csn, abn2, sac, estA, phy, yhcR, bpr and vpr genes as well as enzymatic activities of their products are presented.


Bacilysin biosynthesis by a partially-purified enzyme fraction from Bacillus subtilis
Yazgan, A; Özcengiz, Gülay; Ozcengiz, E; Kilinc, K; Marahiel, MA; Alaeddinoglu, NG (Elsevier BV, 2001-10-04)
Biosynthesis of dipeptide antibiotic bacilysin by a partially purified enzyme prepared from Bacillus subtilis PY79 was studied. Cell material was desintegrated by treatment with lysozyme and sonication and the extract was subjected to ammonium sulfate fractionation. Bacilysin-synthesizing enzyme activity was precipitated between 40% to 70% ammonium sulfate saturation. In vitro enzymatical synthesis of bacilysin was confirmed by performing thin layer chromatographic comparison of the antibiotic formed with t...
Cloning, characterization and heterologous expression of the aspartokinase and aspartate semialdehyde dehydrogenase genes of cephamycin C-producer Streptomyces clavuligerus
Tunca, S; Yilmaz, EI; Piret, J; Liras, P; Özcengiz, Gülay (Elsevier BV, 2004-09-01)
Carbon flow through the lysine branch of the aspartate biosynthetic 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, genes which encode the enzymes catalyzing the first two steps of the aspartate pathway, ask (aspartokinase) and asd (aspartate semialdehyde dehydrogenase), in S. clavuligerus NRRL 3585 were cloned and sequenced. Nucleotide sequencing and codon preference analysis revealed three complete...
Enzymic activity of the K5-type yeast killer toxin and its characterization
Izgu, F; Altinbay, D; Sertkaya, A (Informa UK Limited, 2005-11-01)
K5-type yeast killer toxin secreted by P. anomala NCYC 434 cells has a broad killing spectrum. Competitive inhibiton of killer activity showed that glucans, mainly the beta-1,3 glucan, represent the primary toxin binding site within the cell wall of sensitive cells. Its hydrolytic activity on laminarin in an exo-like fashion revealed that the toxin exerts its killing effect by exo-beta-1,3-glucanase activity. Its specific activity on laminarin was 120U/mg, and the Michaelis constants K-m, and V-max for lami...
Fed-Batch Biomolecule Production by Bacillus subtilis: A State of the Art Review
Ozturk, Sibel; Çalık, Pınar; Ozdamar, Tuncer H. (Elsevier BV, 2016-04-01)
Bacillus subtilis is a highly promising production system for various biomolecules. This review begins with the algorithm of fed-batch operations (FBOs) and then illustrates the approaches to design the initial production medium and/or feed stream. Additionally, the feeding strategies developed with or without feedback control for fed-batch B. subtilis fermentations were compiled with a special emphasis on recombinant protein (r-protein) production. For biomolecule production by wild-type B. subtilis, due t...
Özcengiz, Gülay (Springer Science and Business Media LLC, 1991-01-01)
The dipeptide antibiotic bacilysin, when added externally to the early exponential-phase cultures, markedly limited its own synthesis. It was shown in cell-free extracts that the feedback effect does not involve the inhibition of bacilysin synthetase, the enzyme catalyzing bacilysin formation. We also studied pH and temperature dependence of bacilysin production. Production was highest at about pH 6.8 and at 25-degrees-C.
Citation Formats
G. Özcengiz, E. T. Islerel, and C. Aktas, “Bacillus subtilis overproduces industrially important extracellular enzymes upon the targeted deletion of bacilysin biosynthetic operon,” NEW BIOTECHNOLOGY, pp. 0–0, 2018, Accessed: 00, 2020. [Online]. Available: