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Cellulase cross-linked enzyme aggregates (CLEA) activities can be modulated and enhanced by precipitant selection

Perzon, Alixander; Dicko, Cedric LU ; Çobanoğlu, Özgür; Yükselen, Onur; Eryilmaz, Jitka and Dey, Estera Szwajcer LU (2017) In Journal of Chemical Technology and Biotechnology 92(7). p.1645-1649
Abstract

BACKGROUND: Crosslinked enzyme aggregates (CLEA) technology is a rapid and versatile method to produce immobilized enzymes via precipitation and cross-linking. A direct relationship between CLEA final activity and process parameters is however yet to be clarified. To address the issue, we have used a factorial design to test the formation and optimization of CLEA made from technical grade cellulase (EC 3.2.1.4). Three types of precipitant (ammonium sulfate, polyethylene glycol, tert-butyl alcohol) were used at varied levels of cross-linker concentration, cross-linking time, and temperature. RESULTS: It was found that the CLEAs produced with tert-butyl alcohol were inactive, whereas the polyethylene glycol- and ammonium sulfate-CLEA,... (More)

BACKGROUND: Crosslinked enzyme aggregates (CLEA) technology is a rapid and versatile method to produce immobilized enzymes via precipitation and cross-linking. A direct relationship between CLEA final activity and process parameters is however yet to be clarified. To address the issue, we have used a factorial design to test the formation and optimization of CLEA made from technical grade cellulase (EC 3.2.1.4). Three types of precipitant (ammonium sulfate, polyethylene glycol, tert-butyl alcohol) were used at varied levels of cross-linker concentration, cross-linking time, and temperature. RESULTS: It was found that the CLEAs produced with tert-butyl alcohol were inactive, whereas the polyethylene glycol- and ammonium sulfate-CLEA, recovered 29 and 17% of the free enzyme activity, respectively. Testing for re-usability, we observed that the polyethylene glycol-CLEA maintained 40% of the initial activity after four cycles, in contrast the ammonium sulfate-CLEA only maintained 10% of its activity after one cycle. CONCLUSION: Our study highlights the importance of evaluating the effect of the precipitant on final CLEA activity rather than re-solubilized enzyme activity. It was demonstrated that polyethylene glycol, despite not being able to precipitate the enzymes as readily as ammonium sulfate, resulted in better performing CLEA.

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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
cellulases, cross-linked enzyme aggregates, factorial design, precipitation, recycling
in
Journal of Chemical Technology and Biotechnology
volume
92
issue
7
pages
5 pages
publisher
Wiley-Blackwell
external identifiers
  • scopus:85020270504
  • wos:000403025500020
ISSN
0268-2575
DOI
10.1002/jctb.5160
language
English
LU publication?
yes
id
877c4548-b2e8-4055-801a-0d0323b8ff41
date added to LUP
2017-06-26 15:40:53
date last changed
2018-03-11 04:41:06
@article{877c4548-b2e8-4055-801a-0d0323b8ff41,
  abstract     = {<p>BACKGROUND: Crosslinked enzyme aggregates (CLEA) technology is a rapid and versatile method to produce immobilized enzymes via precipitation and cross-linking. A direct relationship between CLEA final activity and process parameters is however yet to be clarified. To address the issue, we have used a factorial design to test the formation and optimization of CLEA made from technical grade cellulase (EC 3.2.1.4). Three types of precipitant (ammonium sulfate, polyethylene glycol, tert-butyl alcohol) were used at varied levels of cross-linker concentration, cross-linking time, and temperature. RESULTS: It was found that the CLEAs produced with tert-butyl alcohol were inactive, whereas the polyethylene glycol- and ammonium sulfate-CLEA, recovered 29 and 17% of the free enzyme activity, respectively. Testing for re-usability, we observed that the polyethylene glycol-CLEA maintained 40% of the initial activity after four cycles, in contrast the ammonium sulfate-CLEA only maintained 10% of its activity after one cycle. CONCLUSION: Our study highlights the importance of evaluating the effect of the precipitant on final CLEA activity rather than re-solubilized enzyme activity. It was demonstrated that polyethylene glycol, despite not being able to precipitate the enzymes as readily as ammonium sulfate, resulted in better performing CLEA.</p>},
  author       = {Perzon, Alixander and Dicko, Cedric and Çobanoğlu, Özgür and Yükselen, Onur and Eryilmaz, Jitka and Dey, Estera Szwajcer},
  issn         = {0268-2575},
  keyword      = {cellulases,cross-linked enzyme aggregates,factorial design,precipitation,recycling},
  language     = {eng},
  month        = {07},
  number       = {7},
  pages        = {1645--1649},
  publisher    = {Wiley-Blackwell},
  series       = {Journal of Chemical Technology and Biotechnology},
  title        = {Cellulase cross-linked enzyme aggregates (CLEA) activities can be modulated and enhanced by precipitant selection},
  url          = {http://dx.doi.org/10.1002/jctb.5160},
  volume       = {92},
  year         = {2017},
}