Purification and characterization of isocitrate dehydrogenase from Mortierella alpina. (October 2022)

• Record Type: Journal Article
• Authors: Wang, Xuxu; Tang, Xin; Chen, Haiqin; Zhang, Hao; Chen, Yong Q.; Zhao, Jianxin; Chen, Wei
• Is Part Of: Process biochemistry. Volume 121(2022)
• Page Start: 575
• Page End: 583
• Publication Date: 2022-10
• Subjects: MaIDH4 Mortierella alpina Isocitrate dehydrogenase 4 -- NAD+ β-Nicotinamide adenine dinucleotide -- NADP+ β-Nicotinamide adenine dinucleotide phosphate -- NAD-IDH NAD+-dependent isocitrate dehydrogenase -- NADP-IDH NADP+-dependent isocitrate dehydrogenase -- Km Michaelis constant -- kcat catalytic rate constant; Mortierella alpina -- Isocitrate dehydrogenase -- Biochemical characterization -- Kinetics; Biochemical engineering -- Periodicals; Biotechnology -- Periodicals; Biochemistry -- periodicals; Biotechnology -- periodicals; Chemical Engineering -- periodicals; Génie biochimique -- Périodiques; Biotechnologie -- Périodiques; Biochemical engineering; Biotechnology; Periodicals; 660.63
• DOI: 10.1016/j.procbio.2022.07.031
• Languages: English
• ISSNs: 1359-5113

Abstract

Abstract: Isocitrate dehydrogenase (IDH) in oleaginous microorganisms plays an essential role in fatty acid synthesis. We expressed and purified an IDH isozyme from the oleaginous fungus Mortierella alpina ( Ma IDH4) and identified its enzymatic properties. Ma IDH4 is NADP + -dependent and has no detectable activity with NAD + . It exerts its maximum activity when Mn 2+ is used as a cofactor, followed by Mg 2+ . The optimum temperatures was 60 °C and 65 °C, and the optimum pH was 8.5 and 9.5, in the presence of Mn 2+ and Mg 2+, respectively. The catalytic efficiency of Ma IDH4 for NADP + was 16.33 (Mn 2+ ) and 3.86 (Mg 2+ ) μM −1 s −1, while that for isocitrate was 3.06 (Mn 2+ ) and 0.65 (Mg 2+ ) μM −1 s −1 . Characterization of the enzymatic properties of Ma IDH4 provides a basis for further understanding lipid synthesis in M. alpina . Graphical Abstract: ga1 Highlights: Expression of an IDH isozyme from Mortierella alpina ( MaI DH4) in Pichia pastoris. MaI DH4 was purified and characterized. MaI DH4 was thermophilic and basophilic. The optimum metal ion of MaI DH4 was Mn 2+ . MaI DH4 depended only on NADP + .