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  4. Vol 3, Issue 5, 2022

Purification and Characteristics of a NAD(P)-Dependent Gluconate 5-Dehydrogenase from Thermotoga maritima MSB8

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DOI: 10.23977/medsc.2022.030503 | Downloads: 19 | Views: 812

Author(s)

Xiao He 1, Ming Yan 1

Affiliation(s)

1 College of Biotechnology and Pharmaceutical Engineering, Nanjing Tech University, Nanjing, China

Corresponding Author

Xiao He

ABSTRACT

NADP-dependent gluconate 5-dehydrogenase catalyzes the reversible oxidoreduction reaction between D-gluconate and 5-keto-D-gluconate. NADP-dependent gluconate 5-dehydrogenase from bacteria and fungi has been reported. The reported enzyme showed activity only for NADP as the cofactor. This study purified and characterized a NAD(P)-dependent gluconate 5-dehydrogenase (TmGNDH) from the archaea Thermotoga maritima MSB8. TmGNDH is composed of two identical subunits, and a single subunit has 255 amino acids with molecular mass of 28 kDa. TmGNDH is affiliated with the short-chain dehydrogenase/reductase family. Purified TmGNDH has a specific activity of 102.8 U/mg. The enzyme preferred NADP to NAD as its cofactor. TmGNDH exhibited a Km value of 121.3 ± 3.0 mM and k_cat value of 3181 ± 34.1 s^(-1) toward D-gluconate as a substrate. TmGNDH had lower activity with D-sorbitol, D-xylitol, D-glucose and D-xylose as substrates. The optimal pH and temperature of TmGNDH were observed at pH 9.0 and 60℃ with NADP as the cofactor. The thermostability of the enzyme suggests that it can be used as an industrially effective tool for synthesizing 5-keto-D-gluconate.

KEYWORDS

gluconate dehydrogenase, hyperthermophile, archaea, thermostability, 5-keto-D-gluconate

CITE THIS PAPER

Xiao He, Ming Yan, Purification and Characteristics of a NAD(P)-Dependent Gluconate 5-Dehydrogenase from Thermotoga maritima MSB8. MEDS Clinical Medicine (2022) Vol. 3: 12-18. DOI: http://dx.doi.org/10.23977/medsc.2022.030503.

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