An Amperometric Alcohol Sensor Based on Chemically Permeabilized Methylotrophic Microorganisms

Abstract

Chemical permeabilization of the yeast Pichia pastoris NRRL Y‐11430 with 1.0% glycine and 0.1 % Triton X‐100 does not release intracellular alcohol oxidase but alters the cell envelope structure, allowing alcohol oxidase activity to be detected inside the cells (Naglak, 1990). An alcohol sensor based on the use of chemically permeabilized yeast cells was constructed. The sensor response time was within 2 min. The assay time was less than 30s by the initial rate method. The calibration curve for ethanol analysis was linear in the range 5–65 mg/L. The sensor response was almost unaffected by pH changes within the range pH 5–9. This alcohol sensor produced the highest sensitivity for methanol; the sensitivity for ethanol was about half that for methanol. The stability and other characteristics of this new alcohol sensor are also discussed in this work.

Use of the chemical permeabilization process in yeast cells: production of high-activity whole cell biocatalysts

“…Triton X-100, a polyoxyethylene octyl phenyl ether (ionic surfactant), is a chemically inert detergent used in the permeabilization processes of various microorganisms: yeast (Chen et al, 1992;Chow and Palecek, 2004;Kaur and Satyanarayana, 2010), bacteria (Ge et al, 2015;Kinosita et al, 2018;Shin et al, 2016), and fungi (Chelico and Khachatourians, 2003;Villa et al, 2009).…”

“…Triton X-100 has been proven to be effective as a permeabilizing agent in the permeabilization of yeast cells from the genus Pichia and Yarrowia (Table 4) Pichia pastoris Chen et al (1992) Phytase Pichia anomala Kaur and Satyanarayana (2010) Alkaline and acid phosphatase Yarrowia lipolytica Galabova et al (1996) et al (1992) were the first group to prove the effectiveness of Triton X-100 in combination with glycine in the permeabilization process of P. pastoris cells with abundant alcohol oxidase. The compound altered the cell membrane structure, thereby increasing the alcohol oxidase activity, but without causing the enzyme leakage outside the cell.…”