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Characterization of L-rhamnose isomerase from Clostridium stercorarium and its application to the production of D-allose from D-allulose (D-psicose)
- Source :
- Biotechnology letters. 40(2)
- Publication Year :
- 2017
-
Abstract
- To characterize l-rhamnose isomerase (l-RI) from the thermophilic bacterium Clostridium stercorarium and apply it to produce d-allose from d-allulose. A recombinant l-RI from C. stercorarium exhibited the highest specific activity and catalytic efficiency (k cat/K m) for l-rhamnose among the reported l-RIs. The l-RI was applied to the high-level production of d-allose from d-allulose. The isomerization activity for d-allulose was maximal at pH 7, 75 °C, and 1 mM Mn2+ over 10 min reaction time. The half-lives of the l-RI at 65, 70, 75, and 80 °C were 22.8, 9.5, 1.9, and 0.2 h, respectively. To ensure full stability during 2.5 h incubation, the optimal temperature was set at 70 °C. Under the optimized conditions of pH 7, 70 °C, 1 mM Mn2+, 27 U l-RI l−1, and 600 g d-allulose l−1, l-RI from C. stercorarium produced 199 g d-allose l−1 without by-products over 2.5 h, with a conversion yield of 33% and a productivity of 79.6 g l−1 h−1. To the best of our knowledge, this is the highest concentration and productivity of d-allose reported thus far.
- Subjects :
- 0301 basic medicine
Stereochemistry
Bioengineering
Isomerase
Fructose
Applied Microbiology and Biotechnology
Substrate Specificity
03 medical and health sciences
chemistry.chemical_compound
Clostridium
Bacterial Proteins
Enzyme Stability
Escherichia coli
Clostridium stercorarium
Aldose-Ketose Isomerases
L-rhamnose isomerase
biology
Chemistry
Thermophile
Temperature
General Medicine
Hydrogen-Ion Concentration
biology.organism_classification
Recombinant Proteins
030104 developmental biology
Glucose
Yield (chemistry)
Allose
Biotechnology
Subjects
Details
- ISSN :
- 15736776
- Volume :
- 40
- Issue :
- 2
- Database :
- OpenAIRE
- Journal :
- Biotechnology letters
- Accession number :
- edsair.doi.dedup.....8bc02ed3fb27a7fcce27125f6b7484ca