Consistent with the extensive degradation ability on a wide range of macromolecules, the s. The production of regio and stereoselectively hydroxylated steroids is of high pharmaceutical interest and can be achieved by cytochrome p450based biocatalysts. Physical map of the sorangium cellulosum so ce56 chromosome and distribution of cytochrome p450, ferredoxin fdx, and ferredoxin reductase fdr genes. Sequences from three bacteroidetes classes, cytophagia, flavobacteria and sphingobacteria, were distributed into two clades separated by sequences from bacteroidia and some. A bacterial negative transcription regulator binding on an. Users can perform simple and advanced searches based on annotations relating to sequence, structure and function. Sorangium cellulosum so ce56 cells were grown either in m.
Additionally, the corresponding terpene synthases tss. Regio or stereoselective functionalizations of terpenes are one of the main goals of synthetic organic chemistry, which are possible through radical reactions but are not selective enough to introduce the desired chiral alcohol function into those compounds. Other names i polyangium compositum thaxter 1904 sorangium cellulosum imshenetski and solntseva 1936 imshenetski and solntseva 1937 sorangium compositum jahn 1924 sorangium compositum krzemieniewska and krzemieniewski 1937 sorangium nigrescens krzemieniewska and krzemieniewski 1937 more. Microorganisms free fulltext survey of biosynthetic gene. The wild type of the so ce56 strain produces high amounts of chivosazole, under the given. As a model, the crystal structure of cyp260b1 from s. An operon consisting of three open reading frames, annotated in silico as methylmalonylcoa mmcoa epimerase, mmcoa mutase mcm, and meab, was identified in the sequencing project of the myxobacterium sorangium cellulosum so ce56.
Chir was found to serve as a pleiotropic regulator, which not only positively regulated the chivosazol biosynthesis, but also affected the fruiting body development in sorangium cellulosum so ce56. Oct 01, 2010 a novel pyrroloquinoline quinone dependent glucose dehydrogenase like enzyme pqq gdh was isolated from sorangium cellulosum so ce56. Oct 14, 2008 serthrtyr kinases, which together comprise a major class of regulatory proteins in eukaryotes, were not believed to play an important role in prokaryotes until recently. Complete genome sequence of the myxobacterium sorangium. Sorangium cellulosum strain so ce56 is an obligate anaerobic, gramnegative bacterium.
Under stressful conditions this motility, as in other myxobacteria, the cells congregate to form fruiting bodies and differentiate into myxospores. Nitrogen limitation seems to be an important trigger for chivosazol production in s. However, the conversion of progesterone prog by cyp260a1 is very unselective. A novel coldadapted lipase from sorangium cellulosum. Jul 20, 2010 read regioselective hydroxylation of norisoprenoids by cyp109d1 from sorangium cellulosum so ce56, applied microbiology and biotechnology on deepdyve, the largest online rental service for scholarly research with thousands of academic publications available at your fingertips. However, our analysis of 626 prokaryotic genomes reveals that eukaryoticlike protein kinases elks are found in nearly twothirds of the sequenced strains. Regioselective hydroxylation of norisoprenoids by cyp109d1. At the time of this study, complete genome sequence information was available for m. The crystal structure of cyp260a1 consisted of four molecules in the asymmetric unit fig. To test whether poor heterologous expression was the cause of the apparent lack of kmr activity in e. Catalyzes the conversion of 2e,6efarnesyl diphosphate fpp to yield the bicyclic sesquiterpene eremophilene via a 1,10cyclization, which requires the abstraction of the. Sorangium cellulosum is a soildwelling gramnegative bacterium of the group myxobacteria.
Expression of the enzymes and synthesis of their products for nmr analysis and x. The cyp260a1 gene from the previously used plasmid encoding the long protein form was amplified. Ewen,1 dominik pistorius,2 olena perlova,2 norio kagawa,3 alexander o. As p450s are able to perform stereo and regioselective hydroxylation of ster. Genome mining in sorangium cellulosum so ce56 europe pmc. However, it codes only for three type i terpene synthases tss. Terpenoids can be found in almost all forms of life. Complete genome sequence of haliangium ochraceum type strain. A bacterial negative transcription regulator binding on an inverted. In this work, a search for new substrates was performed, combined with product characterization and a structural analysis of substratebound complexes using xray crystallography and computational docking. The novel bacterium shows typical myxobacterial characteristics such as gramnegative, rodshaped vegetative cells, swarming colonies, fruiting bodylike aggregates and bacteriolytic. Sorangium cellulosum so ce56, sorangium cellulosum strain so ce56. Using triparental mating, polyketide synthaseencoding genes were inactivated, leading to chivosazolenegative mutants.
How to analyze and solve genetics problems youtube. As a member of the wwpdb, the rcsb pdb curates and annotates pdb data according to agreed upon standards. The cypome of sorangium cellulosum so ce56 and identification of. Sorangium cellulosum so ce56 is a genomesequenced myxobacterial model strain. Critical variations of conjugational dna transfer into secondary metabolite multiproducing sorangium cellulosum strains so ce12 and so ce56. Automating assessment of the undiscovered biosynthetic. Gene cluster cyp264b1geoa from sorangium cellulosum so ce56. Development of simple media which allow investigations into the global regulation of chivosazol biosynthesis with sorangium cellulosum so ce56. We have identified 2697 elks, most of which are encoded. These molecules are visualized, downloaded, and analyzed by users who range from students to specialized scientists.
Cyp267a1 and cyp267b1 from sorangium cellulosum so ce56 are highly versatile drug metabolizers fredy kern, yogan khatri, martin litzenburger and rita bernhardt department of biochemistry, saarland university, 66123 saarbruecken, germany this article has not been copyedited and formatted. Oct 16, 2009 moreover, the bioinformatic analysis of the so ce56 genome has revealed numerous biosynthetic gene clusters of yet unknown function 11, 12. Sorangium cellulosum strain so ce56,033,779 bp fosmid map of the s. A novel starchdegrading myxobacterium designated noso4t new organism of the sorangiineae strain 4 was isolated in 1995 from a soil sample containing plant residues, collected in lucknow, uttar pradesh, india. The model strain sorangium cellulosum so ce56 possesses a. This work compares the substrate specificities and regioselectivities of the sesquiterpene hydroxylases cyp260a1 and cyp264b1 from myxobacterium sorangium cellulosum so ce56.
Eukaryoticlike protein kinases in the prokaryotes and the. The order myxococcales, a reputable source of biologically active secondary metabolites, spans three suborders which all include natural product producing representatives. Such clusters were created by dproteobacteria and spirochaetes, bacteroidetes and dproteobacterium sorangium cellulosum so ce56, spirochaetes and bacteroidetes. Because of their biotechnological potential as producers of secondary metabolites, the myxobacteria attract attention from both the academic community and the pharmaceutical industry.
The myxobacterium sorangium cellulosum so ce56 is a prolific producer of volatile sesquiterpenes. Very recently, we have shown a very unique 1ahydroxylase activity of c19 steroids by cyp260a1 22. Sorangium cellulosum so ce56 was reported previously. Among actinobacteria, the actinomycetales order actinobacteriadae subclass is the most well. The structures of new terpenes were revealed using nmr, xray diffractometry, polarimetry, and gcms. Characterisation, genome size and genetic manipulation of. These congregating cells make isolation of pure culture and colony counts on agar medium difficult as the bacterium. We report the complete genome sequence of the model sorangium strain s. It has been shown to harbor 21 cytochrome p450 genes. Careful analysis of this genome data revealed that continue reading.
One contig was identified harboring all homologs to the. All 21 so ce56 p450s with respective p450 annotation are assigned in their genomic location of the,033,779 base pairs genome. Sorangium cellulosum so ce56 is a genomesequenced myxobacterial. Mutation in the rel gene of sorangium cellulosum affects. Thuytien ly harvard catalyst profiles harvard catalyst. A single terpene synthase is responsible for a wide. The strain harbours one of the largest prokaryotic genomes.
Comparative genomic analysis of fruiting body formation in. This indicates that substantial parts of genomes are either not of common ancestral origin or have diverged rapidly. Jul 20, 2010 sesquiterpenes are particularly interesting as flavorings and fragrances or as pharmaceuticals. Cyp267a1 and cyp267b1 from sorangium cellulosum so. Microorganisms free fulltext survey of biosynthetic.
The unique hmuy gene sequence as a specific marker of. Based on already characterized p450s, the search for homologous enzymes in databases may also lead to a more targeted. Cyp267b1 is a p450 enzyme from myxobacterium sorangium cellulosum so ce56 displaying a broad substrate scope. Structurebased engineering of steroidogenic cyp260a1 for. Its genome, with 33779 base pairs, is the largest bacterial genome sequenced to date and contains 9367 predicted protein coding genes.
Cyp260a1 from sorangium cellulosum strain so ce56 catalyzes hydroxylation of c19 or c21 steroids at the very unique 1. Sorangium cellulosum strain so ce56 polyangium cellulosum strain so ce56 status. Serthrtyr kinases, which together comprise a major class of regulatory proteins in eukaryotes, were not believed to play an important role in prokaryotes until recently. To validate the assembly, fosmid end sequences were mapped onto the genome sequence by employing the baccardi software1. This study reports the identification and functional characterization of the gene cluster cyp264b1geoa from sorangium cellulosum so ce56. Structure function analysis for the hydroxylation of d c21. This work treats the myxobacterium sorangium cellulosum so ce56 as a producer of volatile terpenes. Genome mining in sorangium cellulosum so ce56 identification and characterization of the homologous electron transfer proteins of a myxobacterial cytochrome p450 kerstin maria ewen. Clicking a gene will select it and show any relevant details. To validate the assembly, fosmid end sequences were. Genome analysis of the fruiting bodyforming myxobacterium. Sixteen genes of strain smp2 t were putatively assigned to the cog category cytoskeleton.
Introduction the emergence of new diseases, rising concerns about drug resistance, and the decreasing efficacy of the existing drugs are of great pharmaceutical concern. Dec 27, 2018 organisms with special secondary metabolism, such as the myxobacterium sorangium cellulosum so ce56, can serve as interesting sources for p450 monooxygenases shimkets et al. Deciphering regulatory mechanisms for secondary metabolite. Metabolic engineering of pseudomonas putida for methylmalonyl. Organisms with special secondary metabolism, such as the myxobacterium sorangium cellulosum so ce56, can serve as interesting sources for p450 monooxygenases shimkets et al. This putative mcm pathway operon was subcloned from a bacterial artificial chromosome by redet recombineering onto a minimal replicon derived from p15a. Ewen km, hannemann f, khatri y, perlova o, kappl r, krug d, huttermann j, muller r, bernhardt r 2009 genome mining in sorangium cellulosum so ce56. Two ferredoxins were able to sustain the catalytic activity of the endogenous s. The first 50 amino acid residues were unresolved because of the absence of the electron. The cypome of sorangium cellulosum so ce56 and identification.
Tof mass spectra for the analysis of peptide mass fingerprints were acquired using a bruker ultraflex toftof mass spectrometer bruker daltonic, bremen, germany and analysed against the s. Unusual outer membrane lipid composition of the gram. We describe the bioinformatic analysis of the so ce56 cytochrome p450 complement consisting of 21 putative p450 genes. Careful analysis of this genome data revealed that part of the genes coding for the very well conserved biosynthesis of lipopolysaccharides lps are missing in this microbe. New sesquiterpene oxidations with cyp260a1 and cyp264b1. The developed assay was able to resolve the effects of crude extracts from s.
To test the developed assays with complex bacterial extracts the cells were incubated with extracts from s. Coinciding with the increase in sequenced bacteria, mining of bacterial genomes for biosynthetic gene clusters bgcs has become a critical component of natural product discovery. The gramnegative myxobacterium sorangium cellulosum so ce56 bears the largest bacterial genome published so far, coding for nearly 10,000 genes. We have identified 2697 elks, most of which are encoded by. The rcsb pdb also provides a variety of tools and resources. Oct 28, 2007 genomewide analysis of direct and palindromic repeats with genalyzer 20 reveals more repetitive sequences in s. As a result, drug research during the past century, driven by chemistry, pharmacology, and clinical science, has. Sorangium cellulosum so ce 56 sorangium cellulosum biocyc. In 1937 imshenetski and solntseva isolated a new species of cellulosedegrading myxobacteria, which they called sorangium cellulosum.
Ntca, a famous global transcriptional regulator responsive to the concentration of nitrogen, was reported to be able to regulate the biosynthesis. Gotermfinderopen source software for accessing gene ontology. Associated with this tremendous biosynthetic power is a high resistance potential against multiple antibiotics. Four tested substrate classes eremophilanes, humulanes, caryophyllanes, and cedranes were converted by both p450s. Dec 22, 2010 the first systematic study of the complete cytochrome p450 complement cypome of sorangium cellulosum so ce56, which is a producer of important secondary metabolites and has the largest bacterial genome sequenced to date, is presented. Researchers reported the complete genome sequence of the model sorangium strain s. The putative coding region was cloned, over expressed in e.1560 724 628 462 917 258 1551 1578 1457 1255 135 379 740 167 976 893 1232 315 992 1426 1237 1255 626 254 9 729 738 1165 421 672