NAD(H) recycling activity of an engineered bifunctional enzyme galactose dehydrogenase/lactate dehydrogenase
(2006) In International Journal of Biological Sciences 2(1). p.6-10- Abstract
A chimeric bifunctional enzyme composing of galactose dehydrogenase (galDH; from Pseudomonas fluorescens) and lactate dehydrogenase (LDH; from Bacillus stearothermophilus) was successfully constructed. The chimeric galDH/LDH possessed dual characteristics of both galactose dehydrogenase and lactate dehydrogenase activities while exhibiting hexameric rearrangement with a molecular weight of approximately 400 kDa. In vitro observations showed that the chimeric enzyme was able to recycle NAD with a continuous production of lactate without any externally added NADH. Two fold higher recycling rate (0.3 mM/h) than that of the native enzyme was observed at pH values above 8.5. Proximity effects became especially pronounced during the recycling... (More)
A chimeric bifunctional enzyme composing of galactose dehydrogenase (galDH; from Pseudomonas fluorescens) and lactate dehydrogenase (LDH; from Bacillus stearothermophilus) was successfully constructed. The chimeric galDH/LDH possessed dual characteristics of both galactose dehydrogenase and lactate dehydrogenase activities while exhibiting hexameric rearrangement with a molecular weight of approximately 400 kDa. In vitro observations showed that the chimeric enzyme was able to recycle NAD with a continuous production of lactate without any externally added NADH. Two fold higher recycling rate (0.3 mM/h) than that of the native enzyme was observed at pH values above 8.5. Proximity effects became especially pronounced during the recycling assay when diffusion hindrance was induced by polyethylene glycol. All these findings open up a high feasibility to apply the NAD(H) recycling system for metabolic engineering purposes e.g. as a model to gain a better understanding on the molecular proximity process and as the routes for synthesizing of numerous high-value-added compounds.
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- author
- Prachayasittikul, Virapong ; Ljung, Sarah ; Isarankura-Na-Ayudhya, Chartchalerm and Bülow, Leif LU
- organization
- publishing date
- 2006
- type
- Contribution to journal
- publication status
- published
- subject
- keywords
- Bacterial Proteins, Enzyme Stability, Escherichia coli, Galactose Dehydrogenases, Geobacillus stearothermophilus, Hydrogen-Ion Concentration, L-Lactate Dehydrogenase, NAD, Protein Engineering, Pseudomonas fluorescens, Recombinant Fusion Proteins
- in
- International Journal of Biological Sciences
- volume
- 2
- issue
- 1
- pages
- 7 pages
- publisher
- Ivyspring International Publisher
- external identifiers
-
- pmid:16585948
- scopus:33646535853
- ISSN
- 1449-2288
- DOI
- 10.7150/ijbs.2.10
- language
- English
- LU publication?
- yes
- id
- aafefe18-d01e-4f62-aa08-4748cc76fcbb
- date added to LUP
- 2016-04-18 15:53:18
- date last changed
- 2024-01-04 02:07:43
@article{aafefe18-d01e-4f62-aa08-4748cc76fcbb,
abstract = {{<p>A chimeric bifunctional enzyme composing of galactose dehydrogenase (galDH; from Pseudomonas fluorescens) and lactate dehydrogenase (LDH; from Bacillus stearothermophilus) was successfully constructed. The chimeric galDH/LDH possessed dual characteristics of both galactose dehydrogenase and lactate dehydrogenase activities while exhibiting hexameric rearrangement with a molecular weight of approximately 400 kDa. In vitro observations showed that the chimeric enzyme was able to recycle NAD with a continuous production of lactate without any externally added NADH. Two fold higher recycling rate (0.3 mM/h) than that of the native enzyme was observed at pH values above 8.5. Proximity effects became especially pronounced during the recycling assay when diffusion hindrance was induced by polyethylene glycol. All these findings open up a high feasibility to apply the NAD(H) recycling system for metabolic engineering purposes e.g. as a model to gain a better understanding on the molecular proximity process and as the routes for synthesizing of numerous high-value-added compounds.</p>}},
author = {{Prachayasittikul, Virapong and Ljung, Sarah and Isarankura-Na-Ayudhya, Chartchalerm and Bülow, Leif}},
issn = {{1449-2288}},
keywords = {{Bacterial Proteins; Enzyme Stability; Escherichia coli; Galactose Dehydrogenases; Geobacillus stearothermophilus; Hydrogen-Ion Concentration; L-Lactate Dehydrogenase; NAD; Protein Engineering; Pseudomonas fluorescens; Recombinant Fusion Proteins}},
language = {{eng}},
number = {{1}},
pages = {{6--10}},
publisher = {{Ivyspring International Publisher}},
series = {{International Journal of Biological Sciences}},
title = {{NAD(H) recycling activity of an engineered bifunctional enzyme galactose dehydrogenase/lactate dehydrogenase}},
url = {{http://dx.doi.org/10.7150/ijbs.2.10}},
doi = {{10.7150/ijbs.2.10}},
volume = {{2}},
year = {{2006}},
}