β-Galactosidase Langmuir Monolayer at Air/X-gal Subphase Interface

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• 作者：Shiv K. Sharma ; Shanghao Li ; Miodrag Micic ; Jhony Orbulescu ; Daniel Weissbart ; Hiromichi Nakahara ; Osamu Shibata ; Roger M. Leblanc
• 刊名：Journal of Physical Chemistry B
• 出版年：2016
• 出版时间：December 8, 2016
• 年：2016
• 卷：120
• 期：48
• 页码：12279-12286
• 全文大小：500K
• ISSN：1520-5207

文摘

This article investigates the surface chemistry properties of the β-galactosidase monolayer at the air-subphase interface at the vicinity of its substrate, X-gal. We have demonstrated that the β-galactosidase in the monolayer form remained active and performed hydrolysis of the X-gal in the subphase. We investigated the β-galactosidase Langmuir monolayer in absence and presence of X-gal in the subphase of varying concentration of X-gal with the sodium chloride solution. It was found that the limiting molecular area as well as the collapse surface pressure kept on decreasing with the increasing concentration of X-gal. In accordance to the data obtained from the isotherm it was also found that β-galactosidase forms a stable monolayer that does not aggregate at the air-subphase interface. The stability of the monolayer at the air-subphase interface was studied by using compression–decompression cycles with and without X-gal at varying concentration and different surface pressures. The infrared reflection–absorption spectroscopy (IRRAS) and Brewster angle microscopy (BAM) of β-galactosidase Langmuir monolayer was also investigated for pure and mixed β-galactosidase at the air-subphase.