Adsorption of biopolyester depolymerase on silicon wafer and poly[(R)-3-hydroxybutyric acid] single crystal revealed by real-time AFM
Adsorption of biopolyester depolymerase on silicon wafer and poly[(R)-3-hydroxybutyric acid] single crystal revealed by real-time AFM
复制标题
DOI：
10.1002/mabi.200500160
复制
发表时间：
2006-01-05
影响因子：
4.6
通讯作者：
Abe, H
中科院分区：
文献类型：
--
作者：
Numata, K;Kikkawa, Y;Abe, H
用实时荧光显微镜和原子力显微镜研究了PHB解聚合酶在硅片和P（3HB）单晶上的吸附行为。首先，对吸附在硅片上的PHB解聚合酶的形貌进行了表征，结果表明，PHB解聚合酶分子的尺寸为2.2 +/- 0.7 nm高，16 +/- 5 nm宽。吸附在P（3HB）单晶上的PHB解聚合酶观察表明，空气中酶在晶体表面的尺寸为1.2 +/- 0.5 nm高，28 +/- 7 nm宽，而在缓冲溶液中检测到的酶分子尺寸为2.1 +/- 0.6 nm高，16 +/- 7 nm宽。空气中PHB解聚合酶与缓冲液中PHB解聚合酶的尺寸比较表明，空气中PHB解聚合酶被压扁，而缓冲液中PHB解聚合酶没有被压扁。此外，还研究了酶吸附对P（3HB）晶体表面分子状态的影响。经酶吸附和乙醇洗涤后的P（3HB）单晶的AFM图像表明，PHB解聚合酶的粘附改变了分子状态，并在晶体表面产生了孔洞。
The adsorption behavior of PHB depolymerase from R. pickettii T1 on a silicon wafer and on P(3HB) single crystals has been studied by real-time and AFM in air and a buffer solution. First, the morphology of PHB depolymerase adsorbed on a silicon wafer was characterized to show that one molecule of PHB depolymerase has dimensions of 2.2 +/- 0.7 nm height and 16 +/- 5 nm width. The observation of PHB depolymerase adsorbed on a P(3HB) single crystal indicated that the dimensions of enzyme on the crystalline surface in air were 1.2 +/- 0.5 nm high and 28 +/- 7 nm wide, while enzyme molecules with dimensions of 2.1 +/- 0.6 nm height and 16 +/- 7 nm width were detected in a buffer solution. Comparison of the dimensions of PHB depolymerase in air with those in a buffer solution showed that the enzyme was squashed in air, but not in a buffer Solution. In addition, the influence of enzymatic adsorption on the molecular state of the P(3HB) crystalline surface was investigated. The AFM images of P(3HB) single crystals after enzymatic adsorption and washing with ethanol indicated that the adhesion of PHB depolymerase changed the molecular state and generated holes on the crystalline surface.