Abstract:
:Acylation of epsilon amino groups of lysyl side chains is a widespread modification of proteins and small molecules in cells of all three domains of life. Recently, we showed that Bacillus subtilis and Bacillus anthracis encode the GCN5-related N-acetyltransferase (GNAT) SatA that can acetylate and inactivate streptothricin, which is a broad-spectrum antibiotic produced by actinomycetes in the soil. To determine functionally relevant residues of B. subtilis SatA (BsSatA), a mutational screen was performed, highlighting the importance of a conserved area near the C terminus. Upon inspection of the crystal structure of the B. anthracis Ames SatA (BaSatA; PDB entry 3PP9), this area appears to form a pocket with multiple conserved aromatic residues; we hypothesized this region contains the streptothricin-binding site. Chemical and site-directed mutagenesis was used to introduce missense mutations into satA, and the functionality of the variants was assessed using a heterologous host (Salmonella enterica). Results of isothermal titration calorimetry experiments showed that residue Y164 of BaSatA was important for binding streptothricin. Results of size exclusion chromatography analyses showed that residue D160 was important for dimerization. Together, these data advance our understanding of how SatA interacts with streptothricin.IMPORTANCE This work provides insights into how an abundant antibiotic found in soil is bound to the enzyme that inactivates it. This work identifies residues for the binding of the antibiotic and probes the contributions of substituting side chains for those in the native protein, providing information regarding hydrophobicity, size, and flexibility of the antibiotic binding site.
journal_name
Appl Environ Microbioljournal_title
Applied and environmental microbiologyauthors
Burckhardt RM,Escalante-Semerena JCdoi
10.1128/AEM.03029-18subject
Has Abstractpub_date
2019-03-06 00:00:00issue
6eissn
0099-2240issn
1098-5336pii
AEM.03029-18journal_volume
85pub_type
杂志文章abstract::Receptors on the cell surfaces of bacterial hosts are essential during the infection cycle of bacteriophages. To date, the phage receptors of the industrial relevant dairy starter bacterium Streptococcus thermophilus remain elusive. Thus, we set out to identify cell surface structures that are involved in host recogni...
journal_title:Applied and environmental microbiology
pub_type: 杂志文章
doi:10.1128/AEM.01847-18
更新日期:2018-11-15 00:00:00
abstract::An organism tentatively identified as Ralstonia eutropha was isolated from enrichment cultures containing tetrahydrofurfuryl alcohol (THFA) as the sole source of carbon and energy. The strain was able to tolerate up to 200 mM THFA in mineral salt medium. The degradation was initiated by an inducible ferricyanide-depen...
journal_title:Applied and environmental microbiology
pub_type: 杂志文章
doi:10.1128/AEM.63.12.4891-4898.1997
更新日期:1997-12-01 00:00:00
abstract::Biobleaching of hardwood unbleached kraft pulp (UKP) by Phanerochaete chrysosporium and Trametes versicolor was studied in the solid-state fermentation system with different culture media. In this fermentation system with low-nitrogen and high-carbon culture medium, pulp brightness increased by 15 and 30 points after ...
journal_title:Applied and environmental microbiology
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journal_title:Applied and environmental microbiology
pub_type: 杂志文章
doi:10.1128/AEM.58.6.1847-1852.1992
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journal_title:Applied and environmental microbiology
pub_type: 杂志文章
doi:10.1128/AEM.40.2.179-185.1980
更新日期:1980-08-01 00:00:00
abstract::An organism capable of growth on pyridine was isolated from soil by enrichment culture techniques and identified as Micrococcus luteus. The organism oxidized pyridine for energy and released N contained in the pyridine ring as ammonium. The organism could not grow on mono- or disubstituted pyridinecarboxylic acids or ...
journal_title:Applied and environmental microbiology
pub_type: 杂志文章
doi:10.1128/AEM.51.5.963-968.1986
更新日期:1986-05-01 00:00:00
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journal_title:Applied and environmental microbiology
pub_type: 杂志文章
doi:10.1128/AEM.61.12.4497-4499.1995
更新日期:1995-12-01 00:00:00
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journal_title:Applied and environmental microbiology
pub_type: 杂志文章
doi:10.1128/aem.66.9.4128-4130.2000
更新日期:2000-09-01 00:00:00
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pub_type: 杂志文章
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更新日期:2014-03-01 00:00:00
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pub_type: 杂志文章
doi:10.1128/AEM.70.10.5825-5832.2004
更新日期:2004-10-01 00:00:00
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pub_type: 杂志文章
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更新日期:2013-11-01 00:00:00
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journal_title:Applied and environmental microbiology
pub_type: 杂志文章
doi:10.1128/AEM.61.9.3293-3298.1995
更新日期:1995-09-01 00:00:00
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journal_title:Applied and environmental microbiology
pub_type: 杂志文章
doi:10.1128/AEM.01095-10
更新日期:2010-10-01 00:00:00
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journal_title:Applied and environmental microbiology
pub_type: 杂志文章
doi:10.1128/AEM.63.10.3946-3949.1997
更新日期:1997-10-01 00:00:00
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journal_title:Applied and environmental microbiology
pub_type: 杂志文章
doi:10.1128/AEM.50.2.287-291.1985
更新日期:1985-08-01 00:00:00
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journal_title:Applied and environmental microbiology
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更新日期:2016-06-30 00:00:00
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journal_title:Applied and environmental microbiology
pub_type: 杂志文章
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更新日期:2012-09-01 00:00:00
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pub_type: 杂志文章,随机对照试验
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journal_title:Applied and environmental microbiology
pub_type: 杂志文章
doi:10.1128/AEM.01739-07
更新日期:2008-04-01 00:00:00
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journal_title:Applied and environmental microbiology
pub_type: 杂志文章
doi:10.1128/AEM.33.2.341-344.1977
更新日期:1977-02-01 00:00:00
abstract::A novel β-(1,3)-glucanase gene designated lamC, cloned from Corallococcus sp. strain EGB, contains a fascin-like module and a glycoside hydrolase family 16 (GH16) catalytic module. LamC displays broad hydrolytic activity toward various polysaccharides. Analysis of the hydrolytic products revealed that LamC is an exo-a...
journal_title:Applied and environmental microbiology
pub_type: 杂志文章
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更新日期:2017-08-01 00:00:00
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journal_title:Applied and environmental microbiology
pub_type: 杂志文章
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更新日期:2005-07-01 00:00:00
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journal_title:Applied and environmental microbiology
pub_type: 杂志文章
doi:10.1128/AEM.00329-19
更新日期:2019-05-16 00:00:00
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journal_title:Applied and environmental microbiology
pub_type: 杂志文章
doi:10.1128/AEM.63.10.3764-3769.1997
更新日期:1997-10-01 00:00:00
abstract::Rhodobacter capsulatus E1F1, a phototrophic purple nonsulfur bacterium capable of photoassimilating nitrate or nitrite, grew phototrophically in the presence of mono- and dinitrophenols with acetate as a carbon source, the highest growth levels being obtained under microaerobic conditions. Utilization of 2,4-dinitroph...
journal_title:Applied and environmental microbiology
pub_type: 杂志文章
doi:10.1128/AEM.58.2.690-695.1992
更新日期:1992-02-01 00:00:00