Discovery of an acidic, thermostable and highly NADP + dependent formate dehydrogenase from Lactobacillus buchneri NRRL B-30929.

Autor: Alpdağtaş S; Department of Biology, Faculty of Science, Van Yuzuncu Yil University, Tusba, 65080, Van, Turkey.; Department of Bioengineering, Faculty of Chemistry and Metallurgy, Yildiz Technical University, Esenler, 34210, Istanbul, Turkey., Yücel S; Department of Bioengineering, Faculty of Chemistry and Metallurgy, Yildiz Technical University, Esenler, 34210, Istanbul, Turkey., Kapkaç HA; Department of Biology, Anadolu University, Tepebaşı, 26470, Eskişehir, Turkey., Liu S; U.S. Department of Agriculture, Renewable Product Technology Research Unit, National Centre for Agricultural Utilization Research, Peoria, IL, USA., Binay B; Department of Bioengineering, Gebze Technical University, Gebze, 41400, Kocaeli, Turkey. binay@gtu.edu.tr.
Jazyk: angličtina
Zdroj: Biotechnology letters [Biotechnol Lett] 2018 Jul; Vol. 40 (7), pp. 1135-1147. Date of Electronic Publication: 2018 May 18.
DOI: 10.1007/s10529-018-2568-6
Abstrakt: Objectives: To identify a robust NADP + dependent formate dehydrogenase from Lactobacillus buchneri NRRL B-30929 (LbFDH) with unique biochemical properties.
Results: A new NADP + dependent formate dehydrogenase gene (fdh) was cloned from genomic DNA of L. buchneri NRRL B-30929. The recombinant construct was expressed in Escherichia coli BL21(DE3) with 6 × histidine at the C-terminus and the purified protein obtained as a single band of approx. 44 kDa on SDS-PAGE and 90 kDa on native-PAGE. The LbFDH was highly active at acidic conditions (pH 4.8-6.2). Its optimum temperature was 60 °C and 50 °C with NADP + and NAD + , respectively and its T m value was 78 °C. Its activity did not decrease after incubation in a solution containing 20% of DMSO and acetonitrile for 6 h. The K M constants were 49.8, 0.12 and 1.68 mM for formate (with NADP + ), NADP + and NAD + , respectively.
Conclusions: An NADP + dependent FDH from L. buchneri NRRL B-30929 was cloned, expressed and identified with its unusual characteristics. The LbFDH can be a promising candidate for NADPH regeneration through biocatalysis requiring acidic conditions and high temperatures.
Databáze: MEDLINE
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