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High-efficiency expression of a superior β-mannanase engineered by cooperative substitution method in Pichia pastoris and its application in preparation of prebiotic mannooligosaccharides
- Source :
- Bioresource Technology. 311:123482
- Publication Year :
- 2020
- Publisher :
- Elsevier BV, 2020.
-
Abstract
- β-mannanase with high specific activity is a prerequisite for the industrial preparation of prebiotic mannooligosaccharides. Three mutants, namely MEI, MER, and MEIR, were constructed by cooperative substitution based on three predominant single-point site mutations (K291E, L211I, and Q112R, respectively). Heterologous expression was facilitated in Pichia pastoris and the recombinase was characterized completely. The specific activities of MER (7481.9 U mg−1) and MEIR (9003.1 U mg−1) increased by 1.07- and 1.29-fold from the initial activity of ME (6970.2U mg−1), respectively. MEIR was used for high-cell-density fermentation to further improve enzyme activity, and the expression levels achieved in the 10-L fermenter were significantly high (105,836 U mL−1). The prebiotic mannooligosaccharides (
- Subjects :
- 0106 biological sciences
Environmental Engineering
medicine.medical_treatment
Mutant
Bioengineering
Industrial fermentation
010501 environmental sciences
01 natural sciences
Pichia
Pichia pastoris
010608 biotechnology
medicine
Recombinase
Waste Management and Disposal
0105 earth and related environmental sciences
biology
Renewable Energy, Sustainability and the Environment
Chemistry
Hydrolysis
Prebiotic
beta-Mannosidase
General Medicine
biology.organism_classification
Enzyme assay
Prebiotics
Biochemistry
biology.protein
Fermentation
Heterologous expression
Amorphophallus
Subjects
Details
- ISSN :
- 09608524
- Volume :
- 311
- Database :
- OpenAIRE
- Journal :
- Bioresource Technology
- Accession number :
- edsair.doi.dedup.....52312707ca57c994a5bcc17d9e027abc