In vitro enzymatic conversion of γ-aminobutyric acid immobilization of glutamate decarboxylase with bacterial cellulose membrane (BCM) and non-linear model establishment

Wanying Yao, Xiao Wu, Jun Zhu, Bo Sun, Curtis Miller

Research output: Contribution to journalArticlepeer-review

24 Scopus citations

Abstract

The work investigated the properties and feasibility of using bacterial cellulose membrane (BCM) as a new and environmental friendly support carrier to immobilize glutamate decarboxylase (GAD) (a unique enzyme in the conversion of γ-aminobutyric acid (GABA) production). During cultivation, the porosities of BCM decreased successively with more extended fibrils piling above one another in a criss-crossing manner thus forming condensed and spatial structure. The BCM with this ultrafine network structure was found to immobilize GAD best via covalent binding because of the highest efficiency of immobilization (87.56% of the enzyme was bonded) and a good operational stability. And the covalent binding efficiency (amount of enzyme immobilized versus lost) was closely related to the porosity or the inner network of the BCM, not to the surface area. The capacity per surface area (mg/cm2) increased from 1.267mg/cm2 to 3.683mg/cm2 when the porosity of BCM ranged from 49% to 73.80%, while a declining trend of the loss of GAD specific activity (from 29.30%/cm2 to 7.38%/cm2) was observed when the porosity increased from 49.9% to 72.30%. Two non-linear regression relationships, between the porosity and loading capacity and between porosity and enzyme activity loss, were empirically modeled with the determination of coefficient R2 of 0.980 and 0.977, respectively. Finally, the established in vitro enzymatic conversion process demonstrated 6.03g/L of GABA at 0.10mol/L Glu, 60min of retention time and 160mL of suspension volume after the 1st run and a loss of 4.15% after the 4th run. The productivity of GABA was 6.03gL-1h-1, higher than that from other reported processes.

Original languageEnglish (US)
Pages (from-to)258-264
Number of pages7
JournalEnzyme and Microbial Technology
Volume52
Issue number4-5
DOIs
StatePublished - Apr 10 2013

Keywords

  • BCM
  • GABA production
  • GAD immobilization
  • In vitro conversion
  • Membrane porosity
  • Non-linear models

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