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1.
Food Chem ; 398: 133863, 2023 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-35961173

RESUMO

Beer quality generally diminishes over time as staling compounds accumulate through various oxidation reactions. Here, we show that refermentation, a traditional practice where Saccharomyces cerevisiae cells are added to beer prior to bottling, diminishes the accumulation of staling aldehydes. However, commonly used beer yeasts only show a limited lifespan in beer. Using high-throughput screening and breeding, we were able to generate novel S. cerevisiae hybrids that survive for over a year in beer. Extensive chemical and sensory analyses of the two most promising hybrids showed that they slow down the accumulation of staling aldehydes, such as furfural and trans-2-nonenal and significantly increased beer flavor stability for up to 12 months. Moreover, the strains did not change the original flavor of the beer, highlighting their potential to be integrated in existing products. Together, these results demonstrate the ability to breed novel microbes that function as natural and sustainable anti-oxidative food preservatives.


Assuntos
Cerveja , Saccharomyces cerevisiae , Aldeídos/análise , Cerveja/análise , Fermentação , Melhoramento Vegetal , Saccharomyces cerevisiae/genética
2.
Microbiology (Reading) ; 162(9): 1620-1628, 2016 09.
Artigo em Inglês | MEDLINE | ID: mdl-27469477

RESUMO

Natural transformation is the main means of horizontal genetic exchange in the obligate human pathogen Neisseria gonorrhoeae and drives the spread of antibiotic resistance and virulence determinants. Transformation can be divided into four steps: (1) DNA binding, (2) DNA uptake, (3) DNA processing and (4) DNA recombination into the chromosome. The DNA processing enzyme DprA has been shown to shuttle incoming ssDNA to the recombination enzyme RecA during transformation in Bacillus subtilis and Streptococcus pneumoniae. Here, we investigate the role of DprA during transformation in N. gonorrhoeae. Inactivation of dprA completely abrogated transformation of gyrB1-encoding DNA, which confers nalidixic acid resistance. The presence of the DNA uptake sequence enhances DNA uptake and transformation by binding to the minor pilus protein ComP. Loss of transformation in the dprA null mutants was independent of the DNA uptake sequence. DprA mutants exhibited increased RecA-dependent pilin variation suggesting that DprA affects pilin variation. Unlike the exquisite UV sensitivity of a recA mutant, inactivation of dprA did not affect survival following UV irradiation. These results demonstrate that DprA has a conserved function during transformation, and reveal additional effects of DprA in N. gonorrhoeae during pilin variation.


Assuntos
Proteínas de Bactérias/metabolismo , Proteínas de Fímbrias/metabolismo , Proteínas de Membrana/metabolismo , Neisseria gonorrhoeae/genética , Neisseria gonorrhoeae/metabolismo , Transformação Bacteriana , Proteínas de Bactérias/genética , DNA Bacteriano/genética , DNA Bacteriano/metabolismo , Proteínas de Fímbrias/genética , Proteínas de Membrana/genética , Recombinases Rec A/genética , Recombinases Rec A/metabolismo
3.
FEMS Microbiol Lett ; 334(1): 44-8, 2012 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-22676068

RESUMO

Natural transformation is the main means of horizontal genetic exchange in the obligate human pathogen Neisseria gonorrhoeae. Neisseria spp. have been shown to preferentially take up and transform their own DNA by recognizing a non-palindromic 10 or 12 nucleotide DNA uptake sequence (DUS10 or DUS12). We investigated the ability of the DUS12 to enhance single-stranded DNA (ssDNA) transformation. Given the non-palindromic nature of the DUS12, we tested whether both strands of the DUS equally enhance transformation. Recombinant single-stranded M13 phage harboring transforming DNA with the Watson DUS12, the Crick DUS12, or no DUS (DUS0) were constructed and circular ssDNA was purified. Southern blots of the purified DNA probed with strand-specific oligonucleotide probes showed > 10,000 : 1 ratio of ssDNA to contaminating dsDNA. The Crick strand of the DUS12 enhanced ssDNA transformation 180- to 470-fold over DUS0 ssDNA, whereas the Watson strand of the DUS only modestly enhanced ssDNA transformation in two strains of N. gonorrhoeae. These data confirm that ssDNA efficiently transforms N. gonorrhoeae, but that there is a strand preference and that part of this strand preference is a greater efficiency of the Crick strand of the DUS12 in enhancing transformation.


Assuntos
DNA Bacteriano/genética , Neisseria gonorrhoeae/genética , Transformação Bacteriana , DNA de Cadeia Simples/genética , Especificidade da Espécie
4.
J Bacteriol ; 192(17): 4436-44, 2010 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-20601472

RESUMO

Natural transformation is the main means of horizontal genetic exchange in the obligate human pathogen Neisseria gonorrhoeae. Neisseria spp. have been shown to preferentially take up and transform their own DNA by recognizing the nonpalindromic 10- or 12-nucleotide sequence 5'-ATGCCGTCTGAA-3' (additional semiconserved nucleotides are underlined), termed the DNA uptake sequence (DUS10 or DUS12). Here we investigated the effects of the DUS on transformation and DNA uptake for several laboratory strains of N. gonorrhoeae. We found that all strains showed efficient transformation of DUS containing DNA (DUS10 and DUS12) but that the level of transformation with DNA lacking a DUS (DUS0) was variable in different strains. The DUS-enhanced transformation was 20-fold in two strains, FA1090 and FA19, but was approximately 150-fold in strains MS11 and 1291. All strains tested provide some level of DUS0 transformation, and DUS0 transformation was type IV pilus dependent. Competition with plasmid DNA revealed that transformation of MS11 was enhanced by the addition of excess plasmid DNA containing a DUS while FA1090 transformation was competitively inhibited. Although FA1090 was able to mediate much more efficient transformation of DNA lacking a DUS than was MS11, DNA uptake experiments showed similar levels of uptake of DNA containing and lacking a DUS in FA1090 and MS11. Finally, DNA uptake was competitively inhibited in both FA1090 and MS11. Taken together, our data indicate that the role of the DUS during DNA transformation is variable between strains of N. gonorrhoeae and may influence multiple steps during transformation.


Assuntos
Sequência de Bases , DNA Bacteriano/metabolismo , Neisseria gonorrhoeae/classificação , Neisseria gonorrhoeae/genética , Transformação Bacteriana , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , DNA Girase/genética , DNA Girase/metabolismo , DNA Bacteriano/química , DNA Bacteriano/genética , Desoxirribonucleases/metabolismo , Fímbrias Bacterianas/genética , Fímbrias Bacterianas/metabolismo , Regulação Bacteriana da Expressão Gênica , Humanos , Neisseria gonorrhoeae/metabolismo , Plasmídeos , Mutação Puntual , Especificidade da Espécie
5.
J Bacteriol ; 192(1): 316-25, 2010 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-19854909

RESUMO

The mismatch correction (MMC) system repairs DNA mismatches and single nucleotide insertions or deletions postreplication. To test the functions of MMC in the obligate human pathogen Neisseria gonorrhoeae, homologues of the core MMC genes mutS and mutL were inactivated in strain FA1090. No mutH homologue was found in the FA1090 genome, suggesting that gonococcal MMC is not methyl directed. MMC mutants were compared to a mutant in uvrD, the helicase that functions with MMC in Escherichia coli. Inactivation of MMC or uvrD increased spontaneous resistance to rifampin and nalidixic acid, and MMC/uvrD double mutants exhibited higher mutation frequencies than any single mutant. Loss of MMC marginally enhanced the transformation efficiency of DNA carrying a single nucleotide mismatch but not that of DNA with a 1-kb insertion. Unlike the exquisite UV sensitivity of the uvrD mutant, inactivating MMC did not affect survival after UV irradiation. MMC and uvrD mutants exhibited increased PilC-dependent pilus phase variation. mutS-deficient gonococci underwent an increased frequency of pilin antigenic variation, whereas uvrD had no effect. Recombination tracts in the mutS pilin variants were longer than in parental gonococci but utilized the same donor pilS loci. These results show that gonococcal MMC repairs mismatches and small insertion/deletions in DNA and also affects the recombination events underlying pilin antigenic variation. The differential effects of MMC and uvrD in gonococci unexpectedly reveal that MMC can function independently of uvrD in this human-specific pathogen.


Assuntos
Variação Antigênica/genética , Reparo de Erro de Pareamento de DNA/fisiologia , Fímbrias Bacterianas/genética , Neisseria gonorrhoeae/genética , Antibacterianos/farmacologia , Proteínas de Bactérias/genética , Proteínas de Bactérias/fisiologia , Reparo de Erro de Pareamento de DNA/genética , Farmacorresistência Bacteriana/genética , Proteínas de Fímbrias/genética , Proteínas de Fímbrias/fisiologia , Teste de Complementação Genética , Resistência a Canamicina/genética , Mutação/genética , Neisseria gonorrhoeae/efeitos dos fármacos , Neisseria gonorrhoeae/efeitos da radiação , Fases de Leitura Aberta/genética , Mutação Puntual , Recombinação Genética/genética , Espectinomicina/farmacologia , Raios Ultravioleta/efeitos adversos
6.
Int J Antimicrob Agents ; 33(4): 321-7, 2009 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-19097863

RESUMO

The aminoglycoside antibiotic kasugamycin (KSG) inhibits translation initiation and thus the growth of many bacteria. In this study, we tested the susceptibilities to KSG of 22 low-passage clinical isolates and 2 laboratory strains of Neisseria gonorrhoeae. Although the range of KSG minimum inhibitory concentrations (MICs) was narrow (seven-fold), clinical isolates and laboratory strains fell into three distinct classes of KSG sensitivity, susceptible, somewhat sensitive and resistant, with MICs of 30, 60-100 and 200 microg/mL, respectively. Two genes have previously been shown to be involved in bacterial KSG resistance: rpsI, which encodes the 30S ribosomal subunit S9 protein; and ksgA, which encodes a predicted dimethyltransferase. Although sequencing of rpsI and ksgA from clinical isolates revealed polymorphisms, none correlated with the MICs of KSG. Ten spontaneous KSG-resistant (KSG(R)) mutants were isolated from laboratory strain FA1090 at a frequency of <4.4x10(-6) resistant colony-forming units (CFU)/total CFU. All isolated KSG(R) variants had mutations in ksgA, whilst no mutations were observed in rpsI. ksgA mutations conferring KSG resistance included four point mutations, two in-frame and one out-of-frame deletions, one in-frame duplication and two frame-shift insertions. These data show a narrow range of susceptibilities for the clinical isolates and laboratory strains examined; moreover, the differences in MICs do not correlate with nucleotide polymorphisms in rpsI or ksgA. Additionally, spontaneous KSG(R) mutants arise by a variety of ksgA mutations.


Assuntos
Aminoglicosídeos/farmacologia , Proteínas de Bactérias/genética , Farmacorresistência Bacteriana , Metiltransferases/genética , Mutação , Neisseria gonorrhoeae/efeitos dos fármacos , Antibacterianos/farmacologia , Análise Mutacional de DNA , DNA Bacteriano/química , DNA Bacteriano/genética , Feminino , Gonorreia/microbiologia , Humanos , Masculino , Testes de Sensibilidade Microbiana , Dados de Sequência Molecular , Neisseria gonorrhoeae/genética , Neisseria gonorrhoeae/isolamento & purificação , Proteínas Ribossômicas/genética , Análise de Sequência de DNA
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