We report the complete genome sequences of two strains of the Alphaproteobacteria genus Rhodobacter, Rhodobacter blasticus 28/5, the source of the commercially available enzyme RsaI, and a new isolate of Rhodobacter sphaeroides 2.4.1. Both strains contain multiple restriction-modification systems, and their DNA methylation motifs are included in this report.
The molecular basis for the intramolecular migration (NIH shift) of the carboxyl group during para-hydroxybenzoate catabolism.
The NIH shift is a chemical rearrangement in which a substituent on an aromatic ring undergoes an intramolecular migration, primarily during an enzymatic hydroxylation reaction. The molecular mechanism for the NIH shift of a carboxyl group has remained a mystery for 40 years. Here, we elucidate the molecular mechanism of the reaction in the conversion of para-hydroxybenzoate (PHB) to gentisate (GA, 2, 5-dihydroxybenzoate). Three genes (phgABC) from the PHB utilizer Brevibacillus laterosporus PHB-7a encode enzymes (p-hydroxybenzoyl-CoA ligase, p-hydroxybenzoyl-CoA hydroxylase and gentisyl-CoA thioesterase, respectively) catalyzing the conversion of PHB to GA via a route involving CoA thioester formation, hydroxylation concomitant with a 1, 2-shift of the acetyl CoA moiety and thioester hydrolysis. The shift of the carboxyl group was established rigorously by stable isotopic experiments with heterologously expressed phgABC, converting 2, 3, 5, 6-tetradeutero-PHB and [carboxyl-13 C]-PHB to 3, 4, 6-trideutero-GA and [carboxyl-13 C]-GA respectively. This is distinct from the NIH shifts of hydrogen and aceto substituents, where a single oxygenase catalyzes the reaction without the involvement of a thioester. The discovery of this three-step strategy for carboxyl group migration reveals a novel role of the CoA thioester in biochemistry and also illustrates the diversity and complexity of microbial catabolism in the carbon cycle.© 2018 John Wiley & Sons Ltd.
The complete genomic sequence of a novel cold-adapted bacterium, Planococcus maritimus Y42, isolated from crude oil-contaminated soil.
Planococcus maritimus Y42, isolated from the petroleum-contaminated soil of the Qaidam Basin, can use crude oil as its sole source of carbon and energy at 20 °C. The genome of P. maritimus strain Y42 has been sequenced to provide information on its properties. Genomic analysis shows that the genome of strain Y42 contains one circular DNA chromosome with a size of 3,718,896 bp and a GC content of 48.8%, and three plasmids (329,482; 89,073; and 12,282 bp). Although the strain Y42 did not show a remarkably higher ability in degrading crude oil than other oil-degrading bacteria, the existence of strain Y42 played a significant role to reducing the overall environmental impact as an indigenous oil-degrading bacterium. In addition, genome annotation revealed that strain Y42 has many genes responsible for hydrocarbon degradation. Structural features of the genomes might provide a competitive edge for P. maritimus strain Y42 to survive in oil-polluted environments and be worthy of further study in oil degradation for the recovery of crude oil-polluted environments.
Complete genome sequence of Pseudomonas plecoglossicida XSDHY-P, a strain that is pathogenic for the marine fish Larimichthys crocea.
Pseudomonas plecoglossicida is a Gram-negative fish pathogen responsi- ble for visceral granular disease in large yellow croaker, Larimichthys crocea. Here, we report the complete genome sequence of a verified virulent strain, XSDHY-P, that was isolated from spleen tissue of a diseased large yellow croaker.
Identification and genome analysis of Deinococcus actinosclerus SJTR1, a novel 17ß-estradiol degradation bacterium.
Biodegradation with microorganisms is considered as an efficient strategy to remove the environmental pollutants. In this work, Deinococcus actinosclerus SJTR1 isolated from the wastewater was confirmed with great degradation capability to 17ß-estradiol, one typical estrogen chemical. It could degrade nearly 90% of 17ß-estradiol (10 mg/L) in 5 days and transform it into estrone; its degradation kinetics fitted for the first-order kinetic equation. The whole genome sequence of D. actinosclerus SJTR1 was obtained and annotated, containing one chromosome (3,315,586 bp) and four plasmids (ranging from 17,267 bp to 460,244 bp). A total of 3913 CDSs and 73 RNA genes (including 12 rRNA genes, 50 tRNA genes, and 11 ncRNA genes) were identified in its whole genome sequence. On this basis, a series of potential genes involved in steroid metabolism and stress responses of D. actinosclerus SJTR1 were predicted. It is the first report of Deinococcus strain with the degradation capability to estrogens. This work could enrich the genome sources of the estrogen-degrading strains and promote the degradation mechanism study of 17ß-estradiol in bacteria.
Complete genome sequence of Bordetella sp. HZ20 sheds light on the ecological role of bacterium without algal-polysaccharides degrading abilities in the brown seaweed-abundant environment
Bordetella sp. HZ20 was isolated from the surface of brown seaweed (Laminaria japonica) and absence of the abilities to decompose the brown seaweed. The genome of Bordetella sp. HZ20 was sequenced and comprised of one circular chromosome with the size of 4,227,194?bp and DNA G?+?C content of 55.5%. Genomic annotation showed that, Bordetella sp. HZ20 may have chitin degradation related enzymes, heparin-sulfate lyase-like protein and enzymes related to the synthase and utilization of polyhydroxyalkanoate for carbon utilization, nitrate and nitrite reductase, glutamate dehydrogenase, glutamate synthase and glutamine synthetase for nitrogen cycle, polyphosphate kinases (pkk1 and pkk2), the high-affinity phosphate-specific transport (Pst) system and the low-affinity inorganic phosphate transporter (pitA) for phosphorus cycle, cysteine synthase and type III acyl coenzyme A transferase (dddD) for sulfur cycle. These features indicated the metabolic patterns of Bordetella sp. HZ20 in C, N, P and S cycles. In addition, the predicted Pst system and cysteine synthase were also related to biofilm formation which showed the potential pathogenicity of Bordetella sp. HZ20 to the cells of animals or plants. This study provides evidences about the metabolic patterns of Bordetella sp. HZ20 and broadens our understandings about ecological roles of bacterium without algal-polysaccharides degrading abilities in the brown seaweed-abundant environment.
Pseudomonas kribbensis is a novel species belonging to the Pseudomonas fluorescens intrageneric group of the genus Pseudomonas. Herein, we report the complete genome sequence of strain 46-2T, isolated from garden soil in Daejeon, South Korea. The 6.32-Mb chromosome contains 5,626 coding sequences with a G+C content of 60.55%.
De novo assembly is the process of reconstructing genomes from DNA fragments (reads), which may contain redundancy and errors. Longer reads simplify assembly and improve contiguity of the output, but current long-read technologies come with high error rates. A crucial step of de novo genome assembly for long reads consists of finding overlapping reads. We present Berkeley Long-Read to Long-Read Aligner and Overlapper (BELLA), which implement a novel approach to compute overlaps using Sparse Generalized Matrix Multiplication (SpGEMM). We present a probabilistic model which demonstrates the soundness of using short, fixed length k-mers to detect overlaps, avoiding expensive pairwise alignment of all reads against all others. We then introduce a notion of reliable k-mers based on our probabilistic model. The use of reliable k-mers eliminates both the k-mer set explosion that would otherwise happen with highly erroneous reads and the spurious overlaps due to k-mers originating from repetitive regions. Finally, we present a new method to separate true alignments from false positives depending on the alignment score. Using this methodology, which is employed in BELLAtextquoterights precise mode, the probability of false positives drops exponentially as the length of overlap between sequences increases. On simulated data, BELLA achieves an average of 2.26% higher recall than state-of-the-art tools in its sensitive mode and 18.90% higher precision than state-of-the-art tools in its precise mode, while being performance competitive.
Genomics and biochemistry investigation on the metabolic pathway of milled wood and alkali lignin-derived aromatic metabolites of Comamonas serinivorans SP-35.
The efficient depolymerization and utilization of lignin are one of the most important goals for the renewable use of lignocelluloses. The degradation and complete mineralization of lignin by bacteria represent a key step for carbon recycling in land ecosystems as well. However, many aspects of this process remain unclear, for example, the complex network of metabolic pathways involved in the degradation of lignin and the catabolic pathway of intermediate aromatic metabolites. To address these subjects, we characterized the deconstruction and mineralization of lignin with milled wood lignin (MWL, the most representative molecule of lignin in its native state) and alkali lignin (AL), and elucidated metabolic pathways of their intermediate metabolites by a bacterium named Comamonas serinivorans SP-35.The degradation rate of MWL reached 30.9%, and its particle size range was decreased from 6 to 30 µm to 2-4 µm-when cultured with C. serinivorans SP35 over 7 days. FTIR analysis showed that the C-C and C-O-C bonds between the phenyl propane structures of lignin were oxidized and cleaved and the side chain structure was modified. More than twenty intermediate aromatic metabolites were identified in the MWL and AL cultures based on GC-MS analysis. Through genome sequencing and annotation, and from GC-MS analysis, 93 genes encoding 33 enzymes and 5 regulatory factors that may be involved in lignin degradation were identified and more than nine metabolic pathways of lignin and its intermediates were predicted. Of particular note is that the metabolic pathway to form the powerful antioxidant 3,4-dihydroxyphenylglycol is described for the first time in bacteria.Elucidation of the ß-aryl ether cleavage pathway in the strain SP-35 indicates that the ß-aryl ether catabolic system is not only present in the family of Sphingomonadaceae, but also other species of bacteria kingdom. These newly elucidated catabolic pathways of lignin in strain SP-35 and the enzymes responsible for them provide exciting biotechnological opportunities for lignin valorization in future.
Paenibacillus bacteria are recovered from varied niches, including human lung, rhizosphere, marine sediments, and hemolymph. Paenibacilli can have plant growth-promoting activities and be antibiotic producers. They can produce exopolysaccharides and enzymes of industrial interest. Illumina and PacBio reads were used to produce a complete genome sequence of the colistin producer Paenibacillus sp. strain B-LR.