Ivanov Alexander E, Thammakhet Chongdee, Kuzimenkova Marina V, Thavarungkul Panote, Kanatharana Proespichaya, Mikhalovska Lyubov I, Mikhalovsky Sergey V, Galaev Igor Yu, Mattiasson Bo
Department of Biotechnology, Center for Chemistry and Chemical Engineering, Lund University, P.O. Box 124, S-22100 Lund, Sweden.
J Mol Recognit. 2008 Mar-Apr;21(2):89-95. doi: 10.1002/jmr.873.
Radical copolymerization of acrylamide (Am) (90 mol%) with N-acryloyl-m-aminophenylboronic acid (NAAPBA) (10 mol%) carried out on the surface of glass slides in aqueous solution and in the absence of chemical cross-linkers, resulted in the formation of thin semitransparent gels. The phenylboronic acid (PBA) ligand density was ca. 160 micromol/ml gel. The gels exhibited a macroporous structure and displayed optical response to sucrose, lactose, glucose and fructose in 50 mM sodium phosphate buffer, in the pH range from 6.5 to 7.5. The response was fairly reversible and linearly depended on glucose concentration in the wide concentration range from 1 to 60 mM at pH 7.3. The character of response was explained by the balance of two competing equilibrium processes: binding of glucose to phenylboronate anions and binary hydrophobic interactions of neutral PBA groups. The apparent diffusion coefficient of glucose in the gels was ca. 2.5 x 10(-7) cm(2)/s. A freshly prepared gel can be used daily for at least 1 month without changes in sensitivity. Autoclaving (121 degrees C, 1.2 bar, 10 min) allows for the gels sterilization, which is important for their use as glucose sensors in fermentation processes.
在水溶液中且无化学交联剂的情况下，于载玻片表面进行丙烯酰胺（Am）（90摩尔%）与N - 丙烯酰基 - m - 氨基苯硼酸（NAAPBA）（10摩尔%）的自由基共聚反应，生成了半透明的薄凝胶。苯基硼酸（PBA）配体密度约为160微摩尔/毫升凝胶。这些凝胶呈现大孔结构，在pH值为6.5至7.5的50 mM磷酸钠缓冲液中对蔗糖、乳糖、葡萄糖和果糖表现出光学响应。该响应相当可逆，并且在pH 7.3时，在1至60 mM的宽浓度范围内线性依赖于葡萄糖浓度。响应特性由两个相互竞争的平衡过程的平衡来解释：葡萄糖与苯基硼酸根阴离子的结合以及中性PBA基团的二元疏水相互作用。葡萄糖在凝胶中的表观扩散系数约为2.5×10⁻⁷厘米²/秒。新制备的凝胶每天至少可使用1个月而灵敏度不变。高压灭菌（121℃，1.2巴，10分钟）可实现凝胶的灭菌，这对于其在发酵过程中用作葡萄糖传感器很重要。
