Production of glucosyl glycerol by immobilized sucrose phosphorylase: Options for enzyme fixation on a solid support and application in microscale flow format

Juan M. Bolivar, Christiane Luley-Goedl, Ernestine Leitner, Thornthan Sawangwan, Bernd Nidetzky

Research output: Contribution to journalArticleResearchpeer-review

Abstract

2-O-(α-D-Glucopyranosyl)-sn-glycerol (αGG) is a natural osmolyte. αGG is produced industrially for application as an active cosmetic ingredient. The biocatalytic process involves a selective transglucosylation from sucrose to glycerol catalyzed by sucrose phosphorylase (SPase). Here we examined immobilization of SPase (from Leuconostoc mesenteroides) on solid support with the aim of enabling continuous production of αGG. By fusing SPase to the polycationic binding module Zbasic2 we demonstrated single-step noncovalent immobilization of the enzyme chimera to different porous supports offering an anionic surface. We showed that immobilization facilitated by Zbasic2 was similarly efficient as immobilization by multipoint covalent attachment on epoxy-activated supports in terms of production of αGG. Enzyme loadings of up to 90 mg enzyme g−1 support were obtained and the immobilized SPase was about half as effective as the enzyme in solution. The high regio- and chemo-selectivity of soluble SPase in αGG synthesis was retained in the immobilized enzyme and product yields of >85% were obtained at titers of ∼800 mM. The Zbasic2-SPase immobilizates were fully recyclable: besides reuse of the enzyme activity, easy recovery of the solid support for fresh immobilizations was facilitated by the reversible nature of the enzyme attachment. Application of immobilized Zbasic2-SPase for continuous production of αGG in a microstructured flow reactor was demonstrated. Space-time yields of 500 mmol αGG L−1 h−1 were obtained at product titers of ∼200 mM. The continuous microreactor was operated for 16 days and an operational half-life of about 10 days was determined.

Original languageEnglish
Pages (from-to)131-138
Number of pages8
JournalJournal of Biotechnology
Volume257
DOIs
Publication statusPublished - 10 Sep 2017

Fingerprint

sucrose phosphorylase
Sugar (sucrose)
Glycerol
Catalyst supports
Enzymes
Immobilization
Immobilized Enzymes
Cosmetics
Enzyme activity
Catalyst selectivity
Half-Life
Sucrose

Keywords

  • 2-O-(α-D-Glucopyranosyl)-sn-glycerol
  • Carbohydrate biotechnology
  • Enzyme microreactors
  • Fusion protein
  • Oriented immobilization
  • Phosphorylase
  • Transglycosylation
  • Z binding module

ASJC Scopus subject areas

  • Biotechnology
  • Applied Microbiology and Biotechnology

Cooperations

  • NAWI Graz

Cite this

Production of glucosyl glycerol by immobilized sucrose phosphorylase : Options for enzyme fixation on a solid support and application in microscale flow format. / Bolivar, Juan M.; Luley-Goedl, Christiane; Leitner, Ernestine; Sawangwan, Thornthan; Nidetzky, Bernd.

In: Journal of Biotechnology, Vol. 257, 10.09.2017, p. 131-138.

Research output: Contribution to journalArticleResearchpeer-review

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