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High level production of a Bacillus amlyoliquefaciens chitosanase in Pichia pastoris suitable for chitooligosaccharides preparation
- Source :
- International Journal of Biological Macromolecules. 149:1034-1041
- Publication Year :
- 2020
- Publisher :
- Elsevier BV, 2020.
-
Abstract
- Chitooligosaccharides (COS) are hydrolytic products of chitosan that are essential in functional food, medicine, and other fields due to their biological activities. Commercial COS are often prepared by the hydrolysis of chitosan by chitosanase. In this study, a glycoside hydrolase family 46 cluster B chitosanase from Bacillus amyloliquefaciens (BaCsn46B) was efficiently expressed in Pichia pastoris. The recombinant enzyme was secreted into the culture medium that reached a total extracellular protein concentration of 4.5 g/L with an activity of 8907.2 U/mL in a high cell density fermenter (5 L). The molecular mass of deglycosylated BaCsn46B was 29.0 kDa. Purified BaCsn46B exhibited excellent enzymatic properties, which had high specific activity (2380.5 U/mg) under optimal reaction conditions (55 °C and pH 6.5). BaCsn46B hydrolyzed chitosan yielded a series of COS with different degrees of polymerization by endo-type cleavage. The end hydrolytic products of BaCsn46B were chitobiose and chitotriose, while no monosaccharide yield was evident in the hydrolytic reaction. The excellent secreted expression level and hydrolytic performance make the enzyme a desirable biocatalyst for the industrial preparation of COS.
- Subjects :
- Glycoside Hydrolases
Bacillus amyloliquefaciens
Oligosaccharides
Bacillus
Chitin
02 engineering and technology
Chitobiose
Biochemistry
Pichia
Pichia pastoris
03 medical and health sciences
chemistry.chemical_compound
Hydrolysis
Structural Biology
Monosaccharide
Glycoside hydrolase
Chitosanase
Molecular Biology
Phylogeny
030304 developmental biology
chemistry.chemical_classification
Chitosan
0303 health sciences
biology
Molecular mass
General Medicine
021001 nanoscience & nanotechnology
biology.organism_classification
Recombinant Proteins
chemistry
Fermentation
0210 nano-technology
Subjects
Details
- ISSN :
- 01418130
- Volume :
- 149
- Database :
- OpenAIRE
- Journal :
- International Journal of Biological Macromolecules
- Accession number :
- edsair.doi.dedup.....fb5c8547bb5155c8c8f3fe2cdd8962c7