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Factors governing the activity of lyophilised and immobilised lipase preparations in organic solvents

Persson, Mattias LU ; Wehtje, Ernst LU and Adlercreutz, Patrick LU (2002) In ChemBioChem 3(6). p.566-571
Abstract
Active site titration and activity measurements were performed in hexane on lyophilised lipase preparations containing different amounts of phosphate buffer and lipase immobilised on porous polypropylene. Lyophilisation of Thermomyces lanuginosus lipase with large quantities of phosphate salts (200 mm) increased the specific activity fourfold, and the number of rapidly titratable active sites increased to 50% from the 73% observed when smaller amounts of phosphate buffer were used (20 mm) during lyophilisation. The phosphate buffer worked as an immobilisation matrix for the lipase, and the increase in specific activity was at least portly due to decreased mass transfer limitations. When lipase was immobilised on porous polypropylene, the... (More)
Active site titration and activity measurements were performed in hexane on lyophilised lipase preparations containing different amounts of phosphate buffer and lipase immobilised on porous polypropylene. Lyophilisation of Thermomyces lanuginosus lipase with large quantities of phosphate salts (200 mm) increased the specific activity fourfold, and the number of rapidly titratable active sites increased to 50% from the 73% observed when smaller amounts of phosphate buffer were used (20 mm) during lyophilisation. The phosphate buffer worked as an immobilisation matrix for the lipase, and the increase in specific activity was at least portly due to decreased mass transfer limitations. When lipase was immobilised on porous polypropylene, the specific activity was 770 times higher than that of the best freeze-dried preparation. At optimal enzyme loading, 93% of the enzyme molecules were titrated at a high rate; this indicates that this adsorption on a hydrophobic surface was a very efficient means of reducing moss transfer limitations and of immobilising the enzyme in its active conformation for use in organic solvents. The variation in specific activity with water activity was found to correlate very well with the variation in titratable active sites when lipases from Burkholderia cepacia and Thermomyces lanuginosus were immobilised on porous polypropylene. The catalytic activity per competent active site was thus constant over the whole range of water activities. (Less)
Please use this url to cite or link to this publication:
author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
water, solvent effects, lipases, enzyme catalysis, immobilisation, activities
in
ChemBioChem
volume
3
issue
6
pages
566 - 571
publisher
John Wiley & Sons
external identifiers
  • wos:000176046400009
  • pmid:12325013
  • scopus:0036905653
ISSN
1439-4227
DOI
10.1002/1439-7633(20020603)3:6<566::AID-CBIC566>3.0.CO;2-7
language
English
LU publication?
yes
id
fab049bd-c262-4649-848e-2b1d6a0652da (old id 336121)
date added to LUP
2007-11-08 14:07:19
date last changed
2017-08-20 03:29:09
@article{fab049bd-c262-4649-848e-2b1d6a0652da,
  abstract     = {Active site titration and activity measurements were performed in hexane on lyophilised lipase preparations containing different amounts of phosphate buffer and lipase immobilised on porous polypropylene. Lyophilisation of Thermomyces lanuginosus lipase with large quantities of phosphate salts (200 mm) increased the specific activity fourfold, and the number of rapidly titratable active sites increased to 50% from the 73% observed when smaller amounts of phosphate buffer were used (20 mm) during lyophilisation. The phosphate buffer worked as an immobilisation matrix for the lipase, and the increase in specific activity was at least portly due to decreased mass transfer limitations. When lipase was immobilised on porous polypropylene, the specific activity was 770 times higher than that of the best freeze-dried preparation. At optimal enzyme loading, 93% of the enzyme molecules were titrated at a high rate; this indicates that this adsorption on a hydrophobic surface was a very efficient means of reducing moss transfer limitations and of immobilising the enzyme in its active conformation for use in organic solvents. The variation in specific activity with water activity was found to correlate very well with the variation in titratable active sites when lipases from Burkholderia cepacia and Thermomyces lanuginosus were immobilised on porous polypropylene. The catalytic activity per competent active site was thus constant over the whole range of water activities.},
  author       = {Persson, Mattias and Wehtje, Ernst and Adlercreutz, Patrick},
  issn         = {1439-4227},
  keyword      = {water,solvent effects,lipases,enzyme catalysis,immobilisation,activities},
  language     = {eng},
  number       = {6},
  pages        = {566--571},
  publisher    = {John Wiley & Sons},
  series       = {ChemBioChem},
  title        = {Factors governing the activity of lyophilised and immobilised lipase preparations in organic solvents},
  url          = {http://dx.doi.org/10.1002/1439-7633(20020603)3:6<566::AID-CBIC566>3.0.CO;2-7},
  volume       = {3},
  year         = {2002},
}