fulltext.study @t Gmail

Optimizing the biocatalytic productivity of an engineered sialidase from Trypanosoma rangeli for 3′-sialyllactose production

Paper ID Volume ID Publish Year Pages File Format Full-Text
17234 42653 2014 9 PDF Available
Title
Optimizing the biocatalytic productivity of an engineered sialidase from Trypanosoma rangeli for 3′-sialyllactose production
Abstract

•Four different enzyme re-use/immobilization methods for a new Trypanosoma rangeli trans-sialidase mutant are examined for 3′-sialyllactose production.•A 9-fold increase in the trans-sialidase biocatalytic productivity for 3′-sialyllactose is achieved by enzyme re-use in a membrane reactor.•The T. rangeli trans-sialidase mutant has a denaturation midpoint temperature (Tm) of 57 °C.

An engineered sialidase, Tr6, from Trypanosoma rangeli was used for biosynthetic production of 3′-sialyllactose, a human milk oligosaccharide case compound, from casein glycomacropeptide (CGMP) and lactose, components abundantly present in industrial dairy side streams. Four different enzyme re-use methods were compared to optimize the biocatalytic productivity, i.e. 3′-sialyllactose formation per amount of Tr6 employed: (i) His-tag immobilization on magnetic Cu2+-iminodiacetic acid-functionalized nanoparticles (MNPs), (ii) membrane immobilization, (iii) calcium alginate encapsulation of cross-linked Tr6, and (iv) Tr6 catalysis in a membrane reactor. Tr6 immobilized on MNPs gave a biocatalytic productivity of 84 mg 3′-sialyllactose/mg Tr6 after seven consecutive reaction runs. Calcium-alginate and membrane immobilization were inefficient. Using free Tr6 in a 10 kDa membrane reactor produced a 9-fold biocatalytic productivity increase compared to using free Tr6 in a batch reactor giving 306 mg 3′-sialyllactose/mg Tr6 after seven consecutive reaction runs. The 3′-sialyllactose yield on α-2,3-bound sialic acid in CGMP was 74%. Using circular dichroism, a temperature denaturation midpoint of Tr6, Tm, of 57.2 °C was determined. The thermal stability of free Tr6 was similarly high and the Tr6 was stable at the reaction temperature (25 °C) for at least 24 h.

Keywords
trans-Sialidase; Trypanosoma rangeli; 3′-Sialyllactose; Enzyme immobilization; Enzymatic membrane reactor; Thermal stability
First Page Preview
Optimizing the biocatalytic productivity of an engineered sialidase from Trypanosoma rangeli for 3′-sialyllactose production
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us
Publisher
Database: Elsevier - ScienceDirect
Journal: Enzyme and Microbial Technology - Volume 55, 5 February 2014, Pages 85–93
Authors
, , , , , ,
Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us