Crystal Structure Analysis and Characterization of NADP-Dependent Glutamate Dehydrogenase with Alcohols Activity from Geotrichum candidum
Source abstract
Abstract
To better understand its mechanism of activity towards higher alcohols, we overexpressed and purified new Geotrichum candidum GDH (GcGDH). The purified GcGDH (50.27 kDa) was then crystallized, and the crystal diffracted to a resolution of 2.3 Å using X-ray diffraction. We found that the GcGDH crystal structure belonged to space group P212121 and was comprised of two hexamers organized into an asymmetric unit, with each subunit consisting of 452 amino acid residues. The binding sites between higher alcohols or L-glutamic acid and GcGDH were consistent. The optimal reaction conditions for GcGDH and hexanol were a pH of 4.0 and temperature of 30 °C, and those for GcGDH and monosodium glutamate (MSG) were a pH of 8.0 and temperature of 20 °C. The Km values for hexanol and MSG were found to be 74.78 mM and 0.018 mM, respectively. Mutating GcGDH Lys 113 to either Ala or Gly caused a dramatic reduction in its catalytic efficiency towards both MSG and hexanol, suggesting that Lys 113 is essential to the active site of GcGDH.
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Research context
OpenAlex topics: Amino Acid Enzymes and Metabolism · Enzyme Structure and Function · Polyamine Metabolism and Applications
SciVX discovery tags are terminology-based. OpenAlex topics are source-provided classifications.
Affiliations
School of Food Science, Xinyang Agriculture and Forestry University, New 24 Street of Yangshan New District, Xinyang 464000, China
Key Laboratory for Space Bioscience and Biotechnology, School of Life Sciences, Northwestern Polytechnical University, 127 Youyi West Road, Xi’an 710072, China
Institute of Medical Research, Northwestern Polytechnical University, 127 Youyi West Road, Xi’an 710072, China
Funding
the Modern Agricultural Industry Technology System · the Natural Science Basic Research Plan in Shaanxi Province of China · the Scientific and Technological Planning in Henan Province · the Foundation for University Key Teacher by the Ministry of Education of Henan · the National Natural Science Foundation of Henan · the High-level research incubator project of College · the Key Research and Development Program of Shaanxi · the Science and Technology Innovation Team of Xinyang Agriculture and Forestry University
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