Glucosylglycerate biosynthesis in the deepest lineage of the bacteria:: Characterization of the thermophilic proteins GpgS and GpgP from Persephonella marina

被引:29
作者
Costa, Joana
Empadinhas, Nuno
da Costa, Milton S. [1 ]
机构
[1] Univ Coimbra, Dept Bioquim, P-3001401 Coimbra, Portugal
[2] Univ Coimbra, Dept Zool, Ctr Neurociencias Biol Celular, P-3004517 Coimbra, Portugal
关键词
D O I
10.1128/JB.00841-06
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
The pathway for the synthesis of glucosylglycerate (GG) in the thermophilic bacterium Persephonella marina is proposed based on the activities of recombinant glucosyl-3-phosphoglycerate (GPG) synthase (GpgS) and glucosyl-3-phosphoglycerate phosphatase (GpgP). The sequences of gpgS and gpgP from the cold-adapted bacterium Methanococcoides burtonii were used to identify the homologues in the genome of P. marina, which were separately cloned and overexpressed as His-tagged proteins in Escherichia coli. The recombinant GpgS protein of P. marina, unlike the homologue from M. burtonii, which was specific for GDP-glucose, catalyzed the synthesis of GPG from UDP-glucose, GDP-glucose, ADP-glucose, and TDP-glucose (in order of decreasing efficiency) and from D-3-phosphoglycerate, with maximal activity at 90 degrees C. The recombinant GpgP protein, like the M. burtonii homologue, dephosphorylated GPG and mannosyl-3-phosphoglycerate (MPG) to GG and mannosylglycerate, respectively, yet at high temperatures the hydrolysis of GPG was more efficient than that of MPG. Gel filtration indicates that GpgS is a dimeric protein, while GpgP is monomeric. This is the first characterization of genes and enzymes for the synthesis of GG in a thermophile.
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页码:1648 / 1654
页数:7
相关论文
共 40 条
[1]   Distribution of genes for synthesis of trehalose and mannosylglycerate in Thermus spp. and direct correlation of these genes with halotolerance [J].
Alarico, S ;
Empadinhas, N ;
Simoes, C ;
Silva, Z ;
Henne, A ;
Mingote, A ;
Santos, H ;
da Costa, MS .
APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 2005, 71 (05) :2460-2466
[2]  
Ames B. N., 1966, METHOD ENZYMOL, V8, P115, DOI DOI 10.1016/0076-6879(66)08014-5
[3]  
[Anonymous], 1989, Molecular Cloning
[4]   Reproduction and metabolism at-10°C of bacteria isolated from Siberian permafrost [J].
Bakermans, C ;
Tsapin, AI ;
Souza-Egipsy, V ;
Gilichinsky, DA ;
Nealson, KH .
ENVIRONMENTAL MICROBIOLOGY, 2003, 5 (04) :321-326
[5]   A histidine-rich metal binding domain at the N terminus of Cu,Zn-superoxide dismutases from pathogenic bacteria [J].
Battistoni, A ;
Pacello, F ;
Mazzetti, AP ;
Capo, C ;
Kroll, JS ;
Langford, PR ;
Sansone, A ;
Donnarumma, G ;
Valenti, P ;
Rotilio, G .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2001, 276 (32) :30315-30325
[6]   Specialized roles of the two pathways for the synthesis of mannosylglycerate in osmoadaptation and thermoadaptation of Rhodothermus marinus [J].
Borges, N ;
Marugg, JD ;
Empadinhas, N ;
da Costa, MS ;
Santos, H .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2004, 279 (11) :9892-9898
[7]  
Brabban AD, 1999, APPL ENVIRON MICROB, V65, P1222
[8]  
BRADFORD MM, 1976, ANAL BIOCHEM, V72, P248, DOI 10.1016/0003-2697(76)90527-3
[9]  
Cánovas D, 1999, APPL ENVIRON MICROB, V65, P3774
[10]   Characterization of the biosynthetic pathway of glucosylglycerate in the archaeon Methanococcoides burtonii [J].
Costa, J ;
Empadinhas, N ;
Gonçalves, L ;
Lamosa, P ;
Santos, H ;
da Costa, MS .
JOURNAL OF BACTERIOLOGY, 2006, 188 (03) :1022-1030