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Asset Tag: Industrial biotechnology

July 7, 2019

Complete genome sequence of a bacterium Pseudomonas fragi P121, a strain with degradation of toxic compounds.

A newly isolated strain P121 was identified as Pseudomonas fragi. The complete genome sequence of P.fragi P121 was carried out using the PacBio RS? platform. The genome contains a circular chromosome with 5,101,809bp. The genome sequence suggests that the P121 exhibited the ability of degradation of toxic compounds. Genome sequencing information provides the genetic basis for the analysis of toxic compounds and the mechanism of extreme environmental adaptation of the strain. Copyright © 2016. Published by Elsevier B.V.

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July 7, 2019

Complete genome sequence of Lactobacillus helveticus CAUH18, a potential probiotic strain originated from koumiss.

Here we report the complete genome sequence of Lactobacillus helveticus CAUH18, a new strain isolated from traditional fermented dairy product koumiss. Its genome has a circular 2.16Mb chromosome with no plasmid. The genome sequence indicated that this strain harbors a gene cluster involved in a novel exopolysaccharides (EPS) biosynthesis and a gene encoding cell-surface aggregation-promoting factors (APFs) to facilitate its colonization in gastrointestinal tract (GIT). This genome sequence provides a basis for further studies about its molecular genetics and probiotic functions. Copyright © 2016. Published by Elsevier B.V.

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July 7, 2019

Structural insights into DNA sequence recognition by Type ISP restriction-modification enzymes.

Engineering restriction enzymes with new sequence specificity has been an unaccomplished challenge, presumably because of the complexity of target recognition. Here we report detailed analyses of target recognition by Type ISP restriction-modification enzymes. We determined the structure of the Type ISP enzyme LlaGI bound to its target and compared it with the previously reported structure of a close homologue that binds to a distinct target, LlaBIII. The comparison revealed that, although the two enzymes use almost a similar set of structural elements for target recognition, the residues that read the bases vary. Change in specificity resulted not only from appropriate substitution of amino acids that contacted the bases but also from new contacts made by positionally distinct residues directly or through a water bridge. Sequence analyses of 552 Type ISP enzymes showed that the structural elements involved in target recognition of LlaGI and LlaBIII were structurally well-conserved but sequentially less-conserved. In addition, the residue positions within these structural elements were under strong evolutionary constraint, highlighting the functional importance of these regions. The comparative study helped decipher a partial consensus code for target recognition by Type ISP enzymes.© The Author(s) 2016. Published by Oxford University Press on behalf of Nucleic Acids Research.

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July 7, 2019

Investigation of proposed ladderane biosynthetic genes from anammox bacteria by heterologous expression in E. coli.

Ladderanes are hydrocarbon chains with three or five linearly concatenated cyclobutane rings that are uniquely produced as membrane lipid components by anammox (anaerobic ammonia-oxidizing) bacteria. By virtue of their angle and torsional strain, ladderanes are unusually energetic compounds, and if produced biochemically by engineered microbes, could serve as renewable, high-energy-density jet fuel components. The biochemistry and genetics underlying the ladderane biosynthetic pathway are unknown, however, previous studies have identified a pool of 34 candidate genes from the anammox bacterium, Kuenenia stuttgartiensis, some or all of which may be involved with ladderane fatty acid biosynthesis. The goal of the present study was to establish a systematic means of testing the candidate genes from K. stuttgartiensis for involvement in ladderane biosynthesis through heterologous expression in E. coli under anaerobic conditions. This study describes an efficient means of assembly of synthesized, codon-optimized candidate ladderane biosynthesis genes in synthetic operons that allows for changes to regulatory element sequences, as well as modular assembly of multiple operons for simultaneous heterologous expression in E. coli (or potentially other microbial hosts). We also describe in vivo functional tests of putative anammox homologs of the phytoene desaturase CrtI, which plays an important role in the hypothesized ladderane pathway, and a method for soluble purification of one of these enzymes. This study is, to our knowledge, the first experimental effort focusing on the role of specific anammox genes in the production of ladderanes, and lays the foundation for future efforts toward determination of the ladderane biosynthetic pathway. Our substantial, but far from comprehensive, efforts at elucidating the ladderane biosynthetic pathway were not successful. We invite the scientific community to take advantage of the considerable synthetic biology resources and experimental results developed in this study to elucidate the biosynthetic pathway that produces unique and intriguing ladderane lipids.

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July 7, 2019

Metabolomics-guided analysis of isocoumarin production by Streptomyces species MBT76 and biotransformation of flavonoids and phenylpropanoids.

Actinomycetes produce the majority of the antibiotics currently in clinical use. The efficiency of antibiotic production is affected by multiple factors such as nutrients, pH, temperature and growth phase. Finding the optimal harvesting time is crucial for successful isolation of the desired bioactive metabolites from actinomycetes, but for this conventional chemical analysis has limitations due to the metabolic complexity. This study explores the utility of NMR-based metabolomics for (1) optimizing fermentation time for the production of known and/or unknown bioactive compounds produced by actinomycetes; (2) elucidating the biosynthetic pathway for microbial natural products; and (3) facilitating the biotransformation of nature-abundant chemicals.

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July 7, 2019

Complete genome sequence of Pseudomonas syringae pv. lapsa strain ATCC 10859, isolated from infected wheat.

Pseudomonas syringae pv. lapsa is a pathovar of Pseudomonas syringae that can infect wheat. The complete genome of P. syringae pv. lapsa strain ATCC 10859 contains a 5,918,899-bp circular chromosome with 4,973 coding sequences, 16 rRNAs, 69 tRNAs, and an average GC content of 59.13%. The analysis of this genome revealed several gene clusters that are related to pathogenesis and virulence. Copyright © 2016 Kong et al.

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July 7, 2019

Complete genome sequence of the engineered Escherichia coli SHuffle strains and their wild-type parents.

SHuffle strains are genetically engineered Escherichia coli strains that are capable of oxidizing cysteines within proteins to form disulfide bonds. Here we present the complete genome of both the K-12 and B versions of SHuffle strains along with their parental ancestors. These strains have been of significant use to both the general scientific community and the biotech industry, interested in producing novel disulfide-bonded proteins that were hitherto unable to be expressed in standard E. coli expression strains.

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July 7, 2019

Complete genome sequence of Hymenobacter sp. strain PAMC26554, an ionizing radiation-resistant bacterium isolated from an Antarctic lichen.

A Gram-negative, rod-shaped, red-pink in color, and UV radiation-resistant bacterium Hymenobacter sp. strain PAMC26554 was isolated from Usnea sp., an Antarctic lichen, and belongs to the class of Cytophagia and the phylum of Bacteroidetes. The complete genome of Hymenobacter sp. PAMC26554 consists of one chromosome (5,244,843bp) with two plasmids (199,990bp and 6421bp). The genomic sequence indicates that Hymenobacter sp. strain PAMC26554 possesses several genes involved in the nucleotide excision repair pathway that protects damaged DNA. This complete genome information will help us to understand its adaptation and novel survival strategy in the Antarctic extreme cold environment. Copyright © 2016 Elsevier B.V. All rights reserved.

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July 7, 2019

Complete genome sequence of the Sporosarcina psychrophila DSM 6497, a psychrophilic Bacillus strain that mediates the calcium carbonate precipitation.

Sporosarcina psychrophila DSM 6497 is a gram positive, spore-formation psychrophilic bacterial strain, widely distributed in terrestrial and aquatic environments. Here we report its complete sequence including one circular chromosome of 4674191bp with a GC content of 40.3%. Genes encoding urease are predicted in the genome, which provide insight information on the microbiologically mediated urea hydrolysis process. This urea hydrolysis can further lead to an increase of carbonate anion and alkalinity in the environment, which promotes the microbiologically induced carbonate precipitation with various applications, such as the bioremediation of calcium rich wastewater and bio-reservation of architectural patrimony. Copyright © 2016 Elsevier B.V. All rights reserved.

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July 7, 2019

Finished genome sequence and methylome of the cyanide-degrading Pseudomonas pseudoalcaligenes strain CECT5344 as resolved by single-molecule real-time sequencing.

Pseudomonas pseudoalcaligenes CECT5344 tolerates cyanide and is also able to utilize cyanide and cyano-derivatives as a nitrogen source under alkaline conditions. The strain is considered as candidate for bioremediation of habitats contaminated with cyanide-containing liquid wastes. Information on the genome sequence of the strain CECT5344 became available previously. The P. pseudoalcaligenes CECT5344 genome was now resequenced by applying the single molecule, real-time (SMRT(®)) sequencing technique developed by Pacific Biosciences. The complete and finished genome sequence of the strain consists of a 4,696,984 bp chromosome featuring a GC-content of 62.34%. Comparative analyses between the new and previous versions of the P. pseudoalcaligenes CECT5344 genome sequence revealed additional regions in the new sequence that were missed in the older version. These additional regions mostly represent mobile genetic elements. Moreover, five additional genes predicted to play a role in sulfoxide reduction are present in the newly established genome sequence. The P. pseudoalcaligenes CECT5344 genome sequence is highly related to the genome sequences of different Pseudomonas mendocina strains. Approximately, 70% of all genes are shared between P. pseudoalcaligenes and P. mendocina. In contrast to P. mendocina, putative pathogenicity genes were not identified in the P. pseudoalcaligenes CECT5344 genome. P. pseudoalcaligenes CECT5344 possesses unique genes for nitrilases and mercury resistance proteins that are of importance for survival in habitats contaminated with cyano- and mercury compounds. As an additional feature of the SMRT sequencing technology, the methylome of P. pseudoalcaligenes was established. Six sequence motifs featuring methylated adenine residues (m6A) were identified in the genome. The genome encodes several methyltransferases, some of which may be considered for methylation of the m6A motifs identified. The complete genome sequence of the strain CECT5344 now provides the basis for exploitation of genetic features for biotechnological purposes. Copyright © 2016 Elsevier B.V. All rights reserved.

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July 7, 2019

Complete genome sequence of the xylan-degrading Mucilaginibacter sp. strain PAMC26640 isolated from an Arctic lichen.

Mucilaginibacter sp. PAMC26640 is a xylan-degrading bacterium isolated from the Arctic lichen Stereocaulon sp. Here, we present the first complete genome sequence of Mucilaginibacter sp. strain PAMC26640, which contains several genes involved in xylan utilization. This genome information provides new insights into the genetic basis of its physiology and further analysis of key metabolic genes related to the xylan degradation pathway. Copyright © 2016. Published by Elsevier B.V.

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July 7, 2019

Complete genome sequence of Pseudomonas azotoformans S4, a potential biocontrol bacterium.

Pseudomonas azotoformans is a Gram-negative bacterium and infects cereal grains, especially rice. P. azotoformans S4 from soil sample derived from Lijiang, Yunnan Province, China, appeared to be strong inhibitory activity against Fusarium fujikurio, a serious rice fungal pathogen. Here, we present the complete genome of P. azotoformans S4, which consists of 6,859,618bp with a circle chromosome, 5991 coding DNA sequences, 70 tRNA and 19 rRNA. The genomic analysis revealed that 9 candidate gene clusters are involved in the biosynthesis of secondary metabolites. Copyright © 2016. Published by Elsevier B.V.

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