title: Protein nanotubes with an enzyme interior surface.
abstract: This report describes the synthesis and enzyme activities of multilayered protein nanotubes with an α-glucosidase (αGluD) interior surface. The nanotubes were prepared by using an alternating layer-by-layer (LbL) assembly of human serum albumin (HSA) and oppositely charged poly-L-arginine (PLA) into a track-etched polycarbonate (PC) membrane (pore size=400 nm) followed by addition of αGluD as the last layer of the wall. Subsequent dissolution of the PC template yielded (PLA/HSA)(2)PLA/αGluD nanotubes. SEM measurements revealed the formation of uniform hollow cylinders with (413±17) nm outer diameter and (52±3) nm wall thickness. In aqueous media, the nanotubes captured a fluorogenic glucopyranoside, 4-methyl-umbelliferyl-α-D-glucopyranoside (MUGlc), into their one-dimensional pore space and hydrolyzed the substrate efficiently to form α-D-glucose. We determined the enzyme parameters (Michaelis constant, K(M), and catalytic constant, k(cat), values) of the protein nanotubes. The several-micrometers-long cylinders were of sufficient length to be spun down by centrifugation at 4000 g, so the product could therefore be easily separated. Similar biocatalysts were prepared by complexation of biotinylated-αGluD into HSA-based nanotubes bearing a single avidin layer as an internal surface. The obtained hybrid nanotubes also exhibited the same enzyme activity for the MUGlc hydrolysis.
authors.affiliation: Department of Applied Chemistry, Faculty of Science and Engineering, Chuo University, 1-13-27 Kasuga, Bunkyo-ku, Tokyo 113-8551, Japan. komatsu@kc.chuo-u.ac.jp
authors.forename: Teruyuki
authors.initials: T
authors.lastname: Komatsu
authors.forename: Hiromi
authors.initials: H
authors.lastname: Terada
authors.forename: Nao
authors.initials: N
authors.lastname: Kobayashi
mesh_terms: D001360:Avidin / Q000737:chemistry*; D005947:Glucose / Q000031:analogs & derivatives* / Q000032:analysis*; D006801:Humans; D015394:Molecular Structure; D043942:Nanotubes / Q000737:chemistry* / Q000648:ultrastructure; D011506:Proteins / Q000737:chemistry*; D012709:Serum Albumin / Q000737:chemistry; D000520:alpha-Glucosidases / Q000737:chemistry* / Q000378:metabolism
chemical_list: D011506:Proteins; D012709:Serum Albumin; D001360:Avidin; D000520:alpha-Glucosidases; D005947:Glucose