• Journal of Microbiology and Biotechnology
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• 제5권4호
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• pp.188-193
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• 1995

A Brevibacterium flavum gene coding for glucose permease of the phosphoenolpyruvate-dependent phosphotransferase system (PTS) was cloned by complementing the Escherichia coli ZSCl13 mutations affecting a ptsG gene with the B. flavum genomic library. From the E. coli clone grown as red colony on a MacConkey plate supplemented with glucose as an additional carbon source, a recombinant plasmid was isolated and named pBFT93. The plasmid pBFT93 was identified as carrying a 3.6-kb fragment of B. flavum chromosomal DNA which enables the E. coli transformant to use glucose or man nose as a sole carbon source in an M9 minimal medium. The non-metabolizable sugar analogues, 2-deoxy-D-glucose (2-DG) and methyl-$\alpha$-D-glucopyranoside (MeGlc) affected the growth of ZSCl13 cells carrying the plasmid pBFT93 on minimal medium supplemented with non-PTS carbohydrate, glycerol, as a sole cabon source, while the analogues did not repress the growth of ZSCl13 cells without pBFT93. It was also found that both $2-deoxy-D-[U-^{14}C]glucose{\;}and{\;}methyl-{\alpha}-D-[U-^{14}C]glucopyranoside$ could be effectively transported into ZSCl13 cells transformed with plasmid pBFT93. Several in vivo complementation studies suggested that the B. flavum DNA in pBFT93 encodes a glucose permease specific for glucose and mannose.

• Journal of Applied Biological Chemistry
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• 제48권4호
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• pp.218-221
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• 2005

Nucleotide sequence of Brevibacterium flavum ptsG gene capable of complementing Escherichia coli ZSC113 mutations defective to glucose permease activity of phosphotransferase system was completely determined, and the gene product was compared with other glucose-specific enzyme II ($EII^{Glc}$). A ptsG gene of B. flavum consisted of open reading frame of 2,025 nucleotides putatively encoding polypeptide of 675 amino acid residues and TAA stop codon. Deduced amino acid sequence of B. flavum ($EII^{Glc}$) had high homology with ($EIIs^{Glc}$) of Corynebacterium glutamicum, C. efficiens, and B. lactofermentum. Arrangement of structural domains, IIBCA, of B. flanum ($EII^{Glc}$) protein was identical to that of EIIs belonging to glucose-phosphotransferase system.

• 한국미생물·생명공학회지
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• 제28권3호
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• pp.147-151
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• 2000

Brevibacterium ammoniagenes 염색체상에서 phosphotrans-ferase system의 glucose permease를 코드하는 ptsG 유전자와 인접한 지역의 염기서열을 결정한 결돠 1,467 nucleo-tides로 구성된 1개의 open reading frame(ORF)이 발견되었고 이것은 489 아미노산 잔기로 구성되는 단백질을 코드하는 것으로추정된다. 이러한 ORF로부터 추정된 단백질의 아미노산 잔기배열을 분석한 결과 30S 리보좀을 구성하는 단백질중의 하나인 S1과 상동성이 높은 것으로 나타났는데 특히 Mycobacterium tuberculosis M. leprae와 Srepto-myces coelicola의 S1단백질의 아미노산 잔기배열과 각각 83%, 74%m, 77%의 매우 높은 상동성을 보였으며 Escherichia coli의 것과도 약 40%의 상동성을 보였다 이로보아 B.ammoniagenes 염색체상에서 ptsG 유전자와 인접한 지역에 존재하는 ORF는 리보좀 단백질 S1의 유전자로 추정된다. 또한 이들은 염색체상에서 동일한 방향으로 판독되며 S1의 유전자가 ptsG의 위 지역으로 266 nucleotides 떨어져 존재하고 있다.

• Journal of Microbiology and Biotechnology
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• 제8권3호
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• pp.214-221
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• 1998

A Brevibacterium ammoniagenes gene coding for glucose/mannose-specific enzyme II ($EII^{Glc}$) of the phosphoenolpyruvate-dependent phosphotransferase system (PTS) was cloned by complementing an Escherichia coli mutation affecting a ptsG gene, and the complete DNA nucleotide sequence was determined. The cloned gene was identified to be a ptsG, which enables the E. coli transportment to use glucose more efficiently than mannose as the sole carbon source in an M9 minimal medium. The ptsG gene of B. ammoniagenes consists of an open reading frame of 1,983 nucleotides putatively encoding a polypeptide of 661 amino acid residues and a TAA stop codon. The deduced amino acid sequence of the B. ammoniagenes $EII^{Glc}$ shows, at $46\%$, the highest degree of sequence similarity with the Corynebacterium glutamicum EII specific for both glucose and mannose. In addition, the $EII^{Glc}$ shares approximately $30\%$ sequence similarities with sucrose-specific and ${\beta}$-glucoside-specific EIIs of the several bacteria belonging to the glucose-PTS class. The 161-amino-acid C-terminal sequence of $EII^{Glc}$ is also similar to that of E. coli enzyme $IIA^{Glc}$, specific for glucose ($EIIA^{Glc}$). The B. ammoniagenes $EII^{Glc}$ consists of three domains; a hydrophobic region (EIIC) and two hydrophilic regions (EIIA, EIIB). The arrangement of structural domains, IIBCA, of the $EII^{Glc}$ is identical to those of EIIs specific for sucrose or ${\beta}$-glucoside. While the domain IIA was removed from the B. ammoniagenes $EII^{Glc}$ the remaining domains IIBC were found to restore the glucose and mannose-utilizing capacity of E. coli mutant lacking $EII^{Glc}$ activity with $EIIA^{Glc}$ of the E. coli mutant. $EII^{Glc}$ contains a histidine residue and a cysteine residue which are putative phosphorylation sites for the protein.

• Journal of Microbiology and Biotechnology
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• 제9권5호
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• pp.582-588
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• 1999

A Brevibacterium lactofermentum gene coding for a glucose-specific permease of the phosphoenolpyruvate-dependent phosphotransferase system (PTS) was cloned, by complementing an Escherichia coli mutation affecting a ptsG gene with the B. lactofermentum genomic library, and completely sequenced. The gene was identified as a ptsG, which enables an E. coli transformant to transport non-metabolizable glucose analogue 2-deoxyglucose (2DG). The ptsG gene of B. lactofermentum consists of an open reading frame of 2,025 nucleotides encoding a polypeptide of 674 amino acid residues and a TAA stop codon. The 3' flanking region contains two stem-loop structures which may be involved in transcriptional termination. The deduced amino acid sequence of the B. lactofermentum enzyme $II^{GIe}$ specific to glucose ($EII^{GIe}$) has a high homology with the Corynebacterium glutamicum enzyme $II^{Man}$ specific to glucose and mannose ($EII^{Man}$), and the Brevibacterium ammoniagenes enzyme $II^{GIc}$ specific to glucose ($EII^{GIc}$). The 171-amino-acid C-terminal sequence of the $EII^{Glc}$ is also similar to the Escherichia coli enzyme $IIA^{GIc}$ specific to glucose ($IIA^{GIc}$). It is interesting that the arrangement of the structural domains, IIBCA, of the B. lactofermentum $EII^{GIc}$ protein is identical to that of EIIs specific to sucrose or $\beta$-glucoside. Several in vivo complementation studies indicated that the B. lactofermentum $EII^{Glc}$ protein could replace both $EII^{ Glc}$ and $EIIA^{Glc}$ in an E. coli ptsG mutant or crr mutant, respectively.

• 미생물학회지
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• 제39권3호
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• pp.141-147
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• 2003

여러 항진균 활성 관련 유전자들의 발현 수준을 동시에 연구하기 위하여 DNA microarray를 이용하여 유전자들의 발현 패턴을 비교 분석하였다. 본 연구에서는 항진균활성을 가지는Bacillus lentimorbus WJ5의 genomic DNA를 무작위 하게 제한효소로 절단하여 2,000개의 DNA단편을 microarray하였으며, 감마선($^{60}Co$)조사로 유도된 7종의 항진균 활성 결핍 돌연변이체와 발현양상을 정량적으로 비교하였다. Gene Cluster (Michael Risen, Stanford Uniy.)를 이용한 DNA microarray의 분석 결과, 총 408개의 DNA 단편이 발현되는 것을 확인할 수 있었으며, 이들 중 20개의 DNA단편이 항진균 활성 결핍 돌연변이체에서 발현이 억제되는 것으로 나타났다. 특히,pbuX (xanthine permease, K222), ywbA (phosphotransferase system enzyme II, K393), ptsG (PTS glucose specific enzyme II ABC component, K877), yufO (ABC transporter(ATP-binding protein), K1301), 그리고 ftsY (signal recognition particle (docking protein), K868)는 모든 돌연변이체에서 동시에 발현되는 down-regulation된 유전자들로서 물질 이동과 관련된 것으로 보고되어 있으며, 항진균 활성 관련 신호 및 물질의 이동에 관여할 것으로 사료되어진다.