Bacteriophages (phages) are natural antibiotics and biological nanoparticles, whose application is significantly boosted by
recent advances of synthetic biology tools. Designer phages are synthetic phages created by genome engineering in a way
to increase the benefits or decrease the drawbacks of natural phages. Here we report the development of a straightforward
genome engineering method to efficiently obtain engineered phages in a model bacterial pathogen, Pseudomonas aeruginosa.
This was achieved by eliminating the wild type phages based on the Streptococcus pyogenes Cas9 (SpCas9) and facilitating
the recombinant generation based on the Red recombination system of the coliphage λ (λRed). The producer (PD) cells of
P. aeruginosa strain PAO1 was created by miniTn7-based chromosomal integration of the genes for SpCas9 and λRed under
an inducible promoter. To validate the efficiency of the recombinant generation, we created the fluorescent phages from a
temperate phage MP29. A plasmid bearing the single guide RNA (sgRNA) gene for selectively targeting the wild type gp35
gene and the editing template for tagging the Gp35 with superfolder green fluorescent protein (sfGFP) was introduced into
the PD cells by electroporation. We found that the targeting efficiency was affected by the position and number of sgRNA.
The fluorescent phage particles were efficiently recovered from the culture of the PD cells expressing dual sgRNA molecules.
This protocol can be used to create designer phages in P. aeruginosa for both application and research purposes.
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Pilin regions that select for the small RNA phages in
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With the growing threat of antibiotic resistance, researchers around the globe are seeking alternatives to stem bacterial
pathogenesis. One such alternative is bacteriocins, proteins produced by bacterial species to inhibit the growth and viability
of related bacterial species. With their diverse mechanisms, which include pore formation and nuclease activities, and
narrow spectrum of activities, which limit their impact to only certain bacterial species, unlike many chemical antibiotics,
bacteriocins offer intriguing possibilities to selectively control individual bacterial populations. Within this review, therefore,
we highlight current research exploring the application of colicins and microcins, a subset of bacteriocins, with an emphasis
on their activities against drug-resistant pathogens, both in in vitro and in vivo settings.
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Isolation, Genomics-Based and Biochemical Characterization of Bacteriocinogenic Bacteria and Their Bacteriocins, Sourced from the Gastrointestinal Tract of Meat-Producing Pigs Ester Sevillano, Irene Lafuente, Nuria Peña, Luis M. Cintas, Estefanía Muñoz-Atienza, Pablo E. Hernández, Juan Borrero International Journal of Molecular Sciences.2024; 25(22): 12210. CrossRef
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Two Gram-stain-positive, rod-shaped, endospore-forming
bacteria, designated 12200R-189T and 14171R-81T were isolated
from the rhizosphere of tomato plants. The 16S rRNA
gene sequence similarity between strains 12200R-189T and
14171R-81T were 97.2%. Both strains showed the highest 16S
rRNA gene sequence similarities to Paenibacillus sacheonensis
SY01T (96.3% and 98.0%, respectively). The genome of strain
12200R-189T was approximately 6.7 Mb in size with 5,750
protein-coding genes (CDSs) and the G + C content was 58.1
mol%, whereas that of strain 14171R-81T comprised one
chromosome of 7.0 Mb and two plasmids (0.2 Mb each) with
6,595 CDSs and the G + C content was 54.5 mol%. Comparative
genome analysis revealed that average nucleotide identity
(ANI) and digital DNA-DNA hybridization (dDDH) values
among 12200R-189T, 14171R-81T, and other closely related
species were below the cut-off levels 95% and 70%, respectively.
Strain 12200R-189T grew at a temperature range
of 15–40°C, pH 6.0–9.0, and 0–3% NaCl (w/v), whereas strain
14171R-81T grew at a temperature range of 10–37°C, pH 6.0–
8.0, and 0–1% NaCl (w/v). Menaquinone-7 (MK-7) was the
only isoprenoid quinone detected in both strains. The predominant
cellular fatty acids (> 10%) were iso-C15:0, anteiso-
C15:0, and iso-C16:0. The polar lipids of strain 12200R-
189T were diphosphatidylglycerol (DPG), phosphatidylglycerol
(PG), phosphatidylethanolamine (PE), aminophospholipid
(APL), phospholipid (PL), phosphatidylglycolipid (PGL),
and four aminophosphoglycolipids (APGLs) and those of
strain 14171R-81T were DPG, PG, PE, APL, three PLs, two
PGLs, and three APGLs. Based on phylogenetic, genomic,
phenotypic, and chemotaxonomic analyses, strains 12200R-
189T and 14171R-81T represent two novel species of the genus
Paenibacillus, for which the names Paenibacillus lycopersici
sp. nov. and Paenibacillus rhizovicinus sp. nov. are proposed.
The type strains are 12200R-189T (= KACC 19916T = CCTCC
AB 2020027T) and 14171R-81T (= KACC 19915T = CCTCC
AB 2020026T).
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Salterns are hypersaline extreme environments with unique
physicochemical properties such as a salinity gradient. Although
the investigation of microbiota in salterns has focused
on archaea and bacteria, diverse fungi also thrive in the brine
and soil of salterns. Fungi isolated from salterns are represented
by black yeasts (Hortaea werneckii, Phaeotheca triangularis,
Aureobasidium pullulans, and Trimmatostroma salinum),
Cladosporium, Aspergillus, and Penicillium species. Most
studies on saltern-derived fungi gave attention to black yeasts
and their physiological characteristics, including growth under
various culture conditions. Since then, biochemical and
molecular tools have been employed to explore adaptation of
these fungi to salt stress. Genome databases of several fungi
in salterns are now publicly available and being used to elucidate
salt tolerance mechanisms and discover the target genes
for agricultural and industrial applications. Notably, the number
of enzymes and novel metabolites known to be produced
by diverse saltern-derived fungi has increased significantly.
Therefore, fungi in salterns are not only interesting and important
subjects to study fungal biodiversity and adaptive
mechanisms in extreme environments, but also valuable bioresources
with potential for biotechnological applications.
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Daqu
Guangsen Fan, Zhilei Fu, Chao Teng, Qiuhua Wu, Pengxiao Liu, Ran Yang, Karim a H M Minhazul, Xiuting Li International Journal of Food Properties.2019; 22(1): 1205. CrossRef
A script for initiating molecular biology studies with non-conventional yeasts based on Saccharomycopsis schoenii Yeseren Kayacan, Adam Griffiths, Jürgen Wendland Microbiological Research.2019; 229: 126342. CrossRef
Designation of rice cake starters for fermented rice products with desired characteristics and fast fermentation Jaruporn Rakmai, Benjamas Cheirsilp, Sirasit Srinuanpan Journal of Food Science and Technology.2019; 56(6): 3014. CrossRef
Overexpression of RAD51 Enables PCR-Based Gene Targeting in Lager Yeast Beatrice Bernardi, Yeseren Kayacan, Madina Akan, Jürgen Wendland Microorganisms.2019; 7(7): 192. CrossRef
Expansion of a Telomeric FLO/ALS-Like Sequence Gene Family in Saccharomycopsis fermentans Beatrice Bernardi, Yeseren Kayacan, Jürgen Wendland Frontiers in Genetics.2018;[Epub] CrossRef
Comparison of volatile and non-volatile metabolites in rice wine fermented by Koji inoculated with Saccharomycopsis fibuligera and Aspergillus oryzae Eun Yeong Son, Sang Mi Lee, Minjoo Kim, Jeong-Ah Seo, Young-Suk Kim Food Research International.2018; 109: 596. CrossRef
Bioformation of Volatile and Nonvolatile Metabolites by Saccharomycopsis fibuligera KJJ81 Cultivated under Different Conditions—Carbon Sources and Cultivation Times Sang Mi Lee, Ji Hye Jung, Jeong-Ah Seo, Young-Suk Kim Molecules.2018; 23(11): 2762. CrossRef