Castañeda M, Guzmán J, Moreno S, Espín G
Departamento de Microbiología Molecular, Instituto de Biotecnología, Universidad Nacional Autónoma de México, Cuernavaca Morelos 62250, México.
J Bacteriol. 2000 May;182(9):2624-8. doi: 10.1128/JB.182.9.2624-2628.2000.
Azotobacter vinelandii produces two polymers: the extracellular polysaccharide alginate and the intracellular polyester poly-beta-hydroxybutyrate (PHB). A cosmid clone (pSMU588) from an A. vinelandii gene library diminished alginate production by A. vinelandii mucoid strain ATCC 9046. The nucleotide sequence and predicted amino acid sequence of the locus responsible for the mucoidy suppression revealed 65% identity to Pseudomonas GacS, a transmembrane sensor kinase of the two-component regulators, whose cognate response regulator, GacA, is a global activator regulating several products and virulence factors. Plasmid pMC15, harboring gacS, and a strain carrying a gacS nonpolar mutation were constructed. Either pMC15 or the gacS mutation significantly reduced alginate production and transcription of algD, the gene coding for the key enzyme GDP-mannose dehydrogenase of the alginate biosynthetic pathway. We found that the gacS mutation also reduced PHB accumulation and impaired encystment. Taken together, these data indicate that in A. vinelandii the gacSA global system regulates polymer synthesis.
胞外多糖藻酸盐和胞内聚酯聚-β-羟基丁酸酯(PHB)。来自维涅兰德固氮菌基因文库的一个黏粒克隆(pSMU588)可减少维涅兰德固氮菌黏液型菌株ATCC 9046的藻酸盐产量。负责抑制黏液形成的基因座的核苷酸序列和预测的氨基酸序列显示,其与铜绿假单胞菌GacS有65%的同一性,GacS是双组分调节因子中的一种跨膜传感激酶,其同源应答调节因子GacA是一种全局激活因子,可调节多种产物和毒力因子。构建了携带gacS的质粒pMC15和携带gacS非极性突变的菌株。pMC15或gacS突变均显著降低了藻酸盐产量以及藻酸盐生物合成途径中关键酶GDP-甘露糖脱氢酶的编码基因algD的转录。我们发现,gacS突变还减少了PHB的积累并损害了包囊形成。综上所述,这些数据表明在维涅兰德固氮菌中,gacSA全局系统调节聚合物合成。