Enzyme dehydration using Microglassification™ preserves the protein's structure and function.

Journal Article (Journal Article)

Controlled enzyme dehydration using a new processing technique of Microglassification™ has been investigated. Aqueous solution microdroplets of lysozyme, α-chymotrypsin, catalase, and horseradish peroxidase were dehydrated in n-pentanol, n-octanol, n-decanol, triacetin, or butyl lactate, and changes in their structure and function were analyzed upon rehydration. Water solubility and microdroplet dissolution rate in each solvent decreased in the order: butyl lactate > n-pentanol > triacetin > n-octanol > n-decanol. Enzymes Microglassified™ in n-pentanol retained higher activity (93%-98%) than n-octanol (78%-85%) or n-decanol (75%-89%), whereas those Microglassified™ in triacetin (36%-75%) and butyl lactate (48%-79%) retained markedly lower activity. FTIR spectroscopy analyses showed α-helix to β-sheet transformation for all enzymes upon Microglassification™, reflecting a loss of bound water in the dried state; however, the enzymes reverted to native-like conformation upon rehydration. Accelerated stressed-storage tests using Microglassified™ lysozyme showed a significant (p < 0.01) decrease in enzymatic activity from 46,560 ± 2736 to 31,060 ± 4327 units/mg after 3 months of incubation; however, it was comparable to the activity of the lyophilized formulation throughout the test period. These results establish Microglassification™ as a viable technique for enzyme preservation without affecting its structure or function.

Full Text

Duke Authors

Cited Authors

  • Aniket, ; Gaul, DA; Bitterfield, DL; Su, JT; Li, VM; Singh, I; Morton, J; Needham, D

Published Date

  • February 2015

Published In

Volume / Issue

  • 104 / 2

Start / End Page

  • 640 - 651

PubMed ID

  • 25557848

Electronic International Standard Serial Number (EISSN)

  • 1520-6017

International Standard Serial Number (ISSN)

  • 0022-3549

Digital Object Identifier (DOI)

  • 10.1002/jps.24279


  • eng