Functional and transcriptional analyses of a fengycin synthetase gene, fenC, from Bacillus subtilis

被引:52
作者
Lin, TP
Chen, CL
Chang, LK
Tschen, JSM
Liu, ST [1 ]
机构
[1] Chang Gung Univ, Dept Microbiol & Immunol, Mol Genet Lab, Tao Yuan 333, Taiwan
[2] Natl Yang Ming Univ, Grad Inst Microbiol & Immunol, Taipei 112, Taiwan
[3] Natl Chung Hsing Univ, Grad Inst Bot, Taichung 402, Taiwan
关键词
D O I
10.1128/JB.181.16.5060-5067.1999
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
A 37-kb DNA fragment containing five fengycin synthetase genes, including fenC, fenD, fenE, fenA, and fenB, was cloned and sequenced. Among these genes,fenC encodes a fengycin synthetase 2,560 amino acids long with an estimated molecular mass of 287 kDa. This protein contains two amino acid activation modules, FenC1 and FenC2, which activate L-glutamic acid and L-ornithine, respectively. Primer extension, using mRNA isolated from the log-phase cells, identified a transcription start site located 86 nucleotides upstream from the initiation codon of fenC, implying that a promoter is located upstream from the start site. Primer extension using total RNA isolated from stationary-phase cells also identified a transcription start site located 61 nucleotides upstream from the initiation codon of fenC. Gene fusion studies demonstrated that in nHA medium, the cells transcribe the fengycin synthetase genes at two different stages of cell growth. The promoter is active during the log phase, and the activity reaches the highest level during the late log phase. The activity decreases sharply but is maintained at a low level for approximately 24 h after cells enter the early stationary phase. The results of this investigation also suggest that the transcription of fenC is positively regulated during the late log phase. Results presented herein provide further insight into fengycin synthesis by B. subtilis F29-3.
引用
收藏
页码:5060 / 5067
页数:8
相关论文
共 43 条
[21]   The complete genome sequence of the Gram-positive bacterium Bacillus subtilis [J].
Kunst, F ;
Ogasawara, N ;
Moszer, I ;
Albertini, AM ;
Alloni, G ;
Azevedo, V ;
Bertero, MG ;
Bessieres, P ;
Bolotin, A ;
Borchert, S ;
Borriss, R ;
Boursier, L ;
Brans, A ;
Braun, M ;
Brignell, SC ;
Bron, S ;
Brouillet, S ;
Bruschi, CV ;
Caldwell, B ;
Capuano, V ;
Carter, NM ;
Choi, SK ;
Codani, JJ ;
Connerton, IF ;
Cummings, NJ ;
Daniel, RA ;
Denizot, F ;
Devine, KM ;
Dusterhoft, A ;
Ehrlich, SD ;
Emmerson, PT ;
Entian, KD ;
Errington, J ;
Fabret, C ;
Ferrari, E ;
Foulger, D ;
Fritz, C ;
Fujita, M ;
Fujita, Y ;
Fuma, S ;
Galizzi, A ;
Galleron, N ;
Ghim, SY ;
Glaser, P ;
Goffeau, A ;
Golightly, EJ ;
Grandi, G ;
Guiseppi, G ;
Guy, BJ ;
Haga, K .
NATURE, 1997, 390 (6657) :249-256
[22]   IDENTIFICATION AND LOCATION OF L-GLYCERATE, AN UNUSUAL ACYL SUBSTITUENT IN GELLAN GUM [J].
KUO, MS ;
MORT, AJ ;
DELL, A .
CARBOHYDRATE RESEARCH, 1986, 156 :173-187
[23]   CLEAVAGE OF STRUCTURAL PROTEINS DURING ASSEMBLY OF HEAD OF BACTERIOPHAGE-T4 [J].
LAEMMLI, UK .
NATURE, 1970, 227 (5259) :680-+
[24]  
Lee S G, 1975, Methods Enzymol, V43, P585
[25]   Molecular cloning and characterization of fengycin synthetase gene fenB from Bacillus subtilis [J].
Li, GH ;
Chen, CL ;
Tschen, JSM ;
Tsay, SS ;
Chang, YS ;
Liu, ST .
JOURNAL OF BACTERIOLOGY, 1998, 180 (05) :1338-1341
[26]   Modular peptide synthetases involved in nonribosomal peptide synthesis [J].
Marahiel, MA ;
Stachelhaus, T ;
Mootz, HD .
CHEMICAL REVIEWS, 1997, 97 (07) :2651-2673
[27]   The tyrocidine biosynthesis operon of Bacillus brevis: Complete nucleotide sequence and biochemical characterization of functional internal adenylation domains [J].
Mootz, HD ;
Marahiel, MA .
JOURNAL OF BACTERIOLOGY, 1997, 179 (21) :6843-6850
[28]   NUCLEOTIDE-SEQUENCES THAT SIGNAL THE INITIATION OF TRANSCRIPTION AND TRANSLATION IN BACILLUS-SUBTILIS [J].
MORAN, CP ;
LANG, N ;
LEGRICE, SFJ ;
LEE, G ;
STEPHENS, M ;
SONENSHEIN, AL ;
PERO, J ;
LOSICK, R .
MOLECULAR & GENERAL GENETICS, 1982, 186 (03) :339-346
[29]  
Sambrook J., 1989, MOL CLONING
[30]   Targeted alteration of the substrate specificity of peptide synthetases by rational module swapping [J].
Schneider, A ;
Stachelhaus, T ;
Marahiel, MA .
MOLECULAR AND GENERAL GENETICS, 1998, 257 (03) :308-318