A new engineered endo-inulinase with improved activity and thermostability: Application in the production of prebiotic fructo-oligosaccharides from inulin

•An engineered endo-inulinase with high activity and thermostability was constructed.•C-terminal fused IBM was effective to improve enzymatic properties.•High temperature in situ inulin hydrolysis was implemented for FOS production.•High purity of FOS (91.4%) were achieved with a productivity of 358...

Full description

Saved in:

Bibliographic Details
Published in:	Food chemistry Vol. 294; pp. 293 - 301
Main Authors:	Mao, Weian, Han, Yaozu, Wang, Xiaoxiang, Zhao, Xiaoxue, Chi, Zhe, Chi, Zhenimg, Liu, Guanglei
Format:	Journal Article
Language:	English
Published:	England Elsevier Ltd 01-10-2019
Subjects:	Bioreactors Engineered endo-inulinase Fructo-oligosaccharides Glycoside Hydrolases - genetics Glycoside Hydrolases - metabolism Hydrogen-Ion Concentration Hydrolysis Inulin Inulin - metabolism Inulin binding module Kinetics Oligosaccharides - metabolism Prebiotic activity Prebiotics Protein Stability Recombinant Proteins - biosynthesis Recombinant Proteins - isolation & purification Saccharomycetales - growth & development Saccharomycetales - metabolism Temperature Glucose (PubChem CID: 5793) 1-Fructofuranosyl-d-Nystose (GF4) (PubChem CID: 123133737) Sucrose (PubChem CID: 5988) Inulin Engineered endo-inulinase Nystose (GF3) (PubChem CID: 166775) Fructo-oligosaccharides Fructose (PubChem CID: 5984) 1-Kestose (GF2) (PubChem CID: 440080) Inulin binding module Prebiotic activity Inulin (PubChem CID: 16219508) Nystose (GF) (PubChem CID: 166775) 1-Fructofuranosyl-d-Nystose (GF) (PubChem CID: 123133737) 1-Kestose (GF) (PubChem CID: 440080)
Online Access:	Get full text
Tags:	Add Tag No Tags, Be the first to tag this record!

Description
Summary:	•An engineered endo-inulinase with high activity and thermostability was constructed.•C-terminal fused IBM was effective to improve enzymatic properties.•High temperature in situ inulin hydrolysis was implemented for FOS production.•High purity of FOS (91.4%) were achieved with a productivity of 358.6 g/L/h. To construct a high-performance engineered endo-inulinase for fructo-oligosaccharides (FOS) production from inulin, an inulin binding module (IBM) was fused into either N- or C-terminal of an endo-inulinase. After heterologous expression, purification and characterization, the C-terminal fusion one (Eninu-IBM) with better activity, thermostability and inulin binding ability was employed for high-temperature in situ inulin hydrolysis in a 10-L fermentor. During this process, Eninu-IBM was first efficiently produced by the yeast cells at 28 °C for 96 h, and subsequently 1600 g unsterilized inulin per liter fermentation liquor was directly supplemented into the bioreactor for FOS production at 60 °C for 2 h. Finally, high purity of FOS (91.4%) were obtained with FOS titer, yield and productivity of 717.3 g/L, 0.912 gFOS/gInulin and 358.6 g/L/h, respectively. The in vitro prebiotic assay indicated that the final FOS products with main polymerization degrees of 3–5 were preferably fermented by beneficial bifidobacteria and lactobacilli.
Bibliography:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23
ISSN:	0308-8146 1873-7072
DOI:	10.1016/j.foodchem.2019.05.062