Chávez-Pacheco J L, Martínez-Yee S, Contreras M L, Gómez-Manzo S, Membrillo-Hernández J, Escamilla J E
Instituto de Fisiología Celular, Universidad Nacional Autonoma de Mexico, Mexico.
J Appl Microbiol. 2005;99(5):1130-40. doi: 10.1111/j.1365-2672.2005.02708.x.
Gluconacetobacter xylinum is well known for its ability to produce large amounts of cellulose, however, little is known about its cell physiology. Our goal was to study the respiratory metabolism and components of the respiratory system of this bacterium in static cultures. To reach our goal, a medium formulation had to be designed to improve cell growth and cellulose production together with a novel method for the recovery of cells from cellulose pellicles.
Successive modifications of a nutrient medium improved G. xylinum cell growth 4.5-fold under static culture conditions. A blender homogenization procedure for the releasing of cells from the cellulose matrix gave a high yield of cells recovered. Respiratory activities of purified cells were greatly stimulated by exogenous substrates and showed to be resistant to KCN. Unexpectedly, exogenous NADH was oxidized at high rates. Cytochromes a, b, c and d were identified after spectral analyses.
Partial bioenergetic characterization of G. xylinum cells allowed us to propose a scheme for its respiratory system. In addition, the growth medium for biomass production and the procedure for the efficient recovery of cells from cellulose pellicles were significantly improved.
This work provides the first-ever bioenergetic characterization of G. xylinum grown in static cultures. In addition, a novel methodology to obtain purified cells in suitable quantities for biochemical research is described.
木醋杆菌以其产生大量纤维素的能力而闻名,然而,对其细胞生理学却知之甚少。我们的目标是研究这种细菌在静态培养中的呼吸代谢和呼吸系统组成部分。为实现这一目标,必须设计一种培养基配方以促进细胞生长和纤维素生成,同时还要开发一种从纤维素薄膜中回收细胞的新方法。
对营养培养基的连续改良使木醋杆菌在静态培养条件下的细胞生长提高了4.5倍。通过搅拌器匀浆程序从纤维素基质中释放细胞,细胞回收率很高。纯化细胞的呼吸活性受到外源底物的极大刺激,并且对氰化钾具有抗性。出乎意料的是,外源NADH被高速氧化。光谱分析后鉴定出细胞色素a、b、c和d。
对木醋杆菌细胞的部分生物能量学特征分析使我们能够提出其呼吸系统的方案。此外,用于生物质生产的生长培养基以及从纤维素薄膜中高效回收细胞的程序得到了显著改进。
这项工作首次对在静态培养中生长的木醋杆菌进行了生物能量学特征分析。此外,还描述了一种获得适量纯化细胞用于生化研究的新方法。