TY - JOUR
T1 - Preparation of cross-linked enzyme aggregates (CLEAs) of an inulosucrase mutant for the enzymatic synthesis of inulin-type fructooligosaccharides
AU - Charoenwongpaiboon, Thanapon
AU - Pichyangkura, Rath
AU - Field, Robert A.
AU - Prousoontorn, Manchumas Hengsakul
N1 - Funding Information:
This research was partially funded by the UK BBSRC Institute Strategic Program on Molecules from Nature (MfN) [BB/PO12523/1] and the John Innes Foundation. The APC was funded by John Innes Centre. Acknowledgments: TC is thankful for scholarship supports from Science Achievement Scholarship of Thailand (SAST), Faculty of Science Chulalongkorn University and Department of Biological Chemistry John Innes Centre.
Funding Information:
Funding: This research was partially funded by the UK BBSRC Institute Strategic Program on Molecules from Nature (MfN) [BB/PO12523/1] and the John Innes Foundation. The APC was funded by John Innes Centre.
Publisher Copyright:
© 2019 by the authors. Licensee MDPI, Basel, Switzerland.
PY - 2019
Y1 - 2019
N2 - Fructooligosaccharides are well-known carbohydrate molecules that exhibit good probiotic activity and are widely used as sweeteners. Inulin-type fructooligosaccharides (IFOs) can be synthesized from sucrose using inulosucrase. In this study, cross-linked enzyme aggregates (CLEAs) of Lactobacillus reuteri 121 inulosucrase (R483A-LrInu) were prepared and used as a biocatalyst for IFOs production. Under optimum conditions, R483A-LrInu CLEAs retained 42% of original inulosucrase activity. Biochemical characterization demonstrated that the optimum pH of inulosucrase changed from 5 to 4 after immobilization, while the optimum temperature was unchanged. Furthermore, the pH stability and thermostability of the R483A-LrInu CLEAs was significantly improved. IFOs product characterization indicated that the product specificity of the enzyme was impacted by CLEA generation, producing a narrower range of IFOs than the soluble enzyme. In addition, the R483A-LrInu CLEAs showed operational stability in the batch synthesis of IFOs.
AB - Fructooligosaccharides are well-known carbohydrate molecules that exhibit good probiotic activity and are widely used as sweeteners. Inulin-type fructooligosaccharides (IFOs) can be synthesized from sucrose using inulosucrase. In this study, cross-linked enzyme aggregates (CLEAs) of Lactobacillus reuteri 121 inulosucrase (R483A-LrInu) were prepared and used as a biocatalyst for IFOs production. Under optimum conditions, R483A-LrInu CLEAs retained 42% of original inulosucrase activity. Biochemical characterization demonstrated that the optimum pH of inulosucrase changed from 5 to 4 after immobilization, while the optimum temperature was unchanged. Furthermore, the pH stability and thermostability of the R483A-LrInu CLEAs was significantly improved. IFOs product characterization indicated that the product specificity of the enzyme was impacted by CLEA generation, producing a narrower range of IFOs than the soluble enzyme. In addition, the R483A-LrInu CLEAs showed operational stability in the batch synthesis of IFOs.
KW - Cross-linked enzyme aggregates
KW - Fructooilgosaccharide
KW - Inulosucrase
UR - http://www.scopus.com/inward/record.url?scp=85070620468&partnerID=8YFLogxK
U2 - 10.3390/catal9080641
DO - 10.3390/catal9080641
M3 - Article
AN - SCOPUS:85070620468
VL - 9
JO - Catalysts
JF - Catalysts
SN - 2073-4344
IS - 8
M1 - 641
ER -