Enzymatic hydrolysis of nylons: quantification of the reaction rate of nylon hydrolase for thin-layered nylons

• Published in: Applied microbiology and biotechnology Vol. 98; no. 20; pp. 8751 - 8761
• Main Authors: Nagai, Keisuke, Iida, Kazuki, Shimizu, Kimiaki, Kinugasa, Ryo, Izumi, Motoki, Kato, Dai-ichiro, Takeo, Masahiro, Mochiji, Kozo, Negoro, Seiji
• Format: Journal Article
• Language: English
• Published: Berlin/Heidelberg Springer-Verlag 01.10.2014; Springer Berlin Heidelberg; Springer; Springer Nature B.V
• Subjects: Analysis; Assaying; Biodegradation; Biomedical and Life Sciences; Biopolymers; Biotechnology; Cellulose; Chemical properties; Chloride; Chromatography; Chromatography, Thin Layer; Copolymers; Degradation; Engineering schools; Environmental Biotechnology; enzymatic hydrolysis; Enzymes; Hydrolases - metabolism; Hydrolysis; image analysis; Laboratories; Life Sciences; Microbial Genetics and Genomics; Microbiology; Mutation; Nylon; Nylons; Nylons - metabolism; Optical Imaging; Polyamide resins; Polyamides; Polymerization; Polymers; Studies; Temperature; Time Factors; Japan; Biodegradation; Nylon-66; Copolymer; Nylon-64; Nylon hydrolase; Nylon-6
• ISSN: 0175-7598; 1432-0614
• DOI: 10.1007/s00253-014-5885-2

Nylon hydrolase degrades various aliphatic nylons, including nylon-6 and nylon-66. We synthesized a nylon-66 copolymer (M w = 22,900, M ₙ = 7,400), in which a part of an adipoyl unit (32 % molar ratio) of nylon-66 was replaced with a succinyl unit by interfacial polymerization. To quantify the reaction rate of the enzymatic hydrolysis of nylons at the surface of solid polymers, we prepared a thin layer of nylons on the bottom surface of each well in a polystyrene-based micro-assay plate. The thickness of the nylon layer was monitored by imaging analysis of the photographic data. More than 99 % of the copolymer with thicknesses of 260 nm (approximately 600 layers of polymer strands) were converted to water-soluble oligomers by nylon hydrolase (3 mg enzyme ml⁻¹) at 30 °C within 60 h. These results were further confirmed by TLC analysis of the reaction products and by assay of liberated amino groups in the soluble fractions. The degradation rate of the thin-layered nylon-6 was similarly analyzed. We demonstrate that this assay enables a quantitative evaluation of the reaction rate of hydrolysis at the interface between the solid and aqueous phases and a quantitative comparison of the degradability for various polyamides.