Rombola Tiago Henrique, Pedrinho Eliamar Aparecida Nascimbem, de Macedo Lemos Eliana Gertrudes, Gonçalves Adriano Marques, dos Santos Luiz Flávio José, Pizauro João Martins
Faculdade de Ciências Agrárias e Veterinárias (FCAV), UNESP - Univ Estadual Paulista, Câmpus de Jaboticabal, Departamento de Tecnologia, Laboratório de Enzimologia Aplicada, Jaboticabal, SP, Brazil.
BMC Res Notes. 2014 Apr 9;7:221. doi: 10.1186/1756-0500-7-221.
The genus Burkholderia is widespread in diverse ecological niches, the majority of known species are soil bacteria that exhibit different types of non-pathogenic interactions with plants. Burkholderia species are versatile organisms that solubilize insoluble minerals through the production of organic acids, which increase the availability of nutrients for the plant. Therefore these bacteria are promising candidates for biotechnological applications.
Burkholderia sp. (R 3.25 isolate) was isolated from agricultural soil in Ponta Grossa-PR-Brazil and identified through analysis of the 16S rDNA as a strain classified as Burkholderia gladioli. The expression of membrane-bound acid phosphatase (MBAcP) was strictly regulated with optimal expression at a concentration of phosphorus 5 mM. The apparent optimum pH for the hydrolysis of p-nitrophenylphosphate (PNPP) was 6.0. The hydrolysis of PNPP by the enzyme exhibited a hyperbolic relationship with increasing concentration of substrate and no inhibition by excess of substrate was observed. Kinetic data revealed that the hydrolysis of PNPP exhibited cooperative kinetics with n = 1.3, Vm = 113.5 U/mg and K0.5 = 65 μM. The PNPPase activity was inhibited by vanadate, p-hydroxymercuribenzoate, arsenate and phosphate, however the activity was not inhibited by calcium, levamisole, sodium tartrate, EDTA, zinc, magnesium, cobalt, ouabain, oligomycin or pantoprazol.
The synthesis of membrane-bound non-specific acid phosphatase, strictly regulated by phosphate, and its properties suggest that this bacterium has a potential biotechnological application to solubilize phosphate in soils with low levels of this element, for specific crops.
伯克霍尔德氏菌属广泛存在于各种生态位中，已知的大多数物种是土壤细菌，它们与植物表现出不同类型的非致病相互作用。伯克霍尔德氏菌属的物种是多功能生物，通过产生有机酸使不溶性矿物质溶解，从而增加植物可利用的养分。因此，这些细菌是生物技术应用的有前途的候选者。
伯克霍尔德氏菌（R 3.25分离株）从巴西巴拉那州蓬塔格罗萨的农业土壤中分离出来，并通过16S rDNA分析鉴定为伯克霍尔德氏唐菖蒲菌株。膜结合酸性磷酸酶（MBAcP）的表达受到严格调控，在磷浓度为5 mM时表达最佳。对硝基苯磷酸酯（PNPP）水解的表观最适pH为6.0。该酶对PNPP的水解与底物浓度增加呈双曲线关系，未观察到底物过量的抑制作用。动力学数据表明，PNPP的水解表现出协同动力学，n = 1.3，Vm = 113.5 U/mg，K0.5 = 65 μM。PNPPase活性受到钒酸盐、对羟基汞苯甲酸、砷酸盐和磷酸盐的抑制，然而该活性不受钙、左旋咪唑、酒石酸钠、EDTA、锌、镁、钴、哇巴因、寡霉素或泮托拉唑的抑制。
膜结合非特异性酸性磷酸酶的合成受磷酸盐严格调控，其特性表明该细菌在生物技术方面具有潜在应用，可用于特定作物，在低磷土壤中溶解磷。
