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l-myo-Inositol-1-Phosphate Synthase Expressed in Developing Organ: Isolation and Characterisation of the Enzyme from Human Fetal Liver

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Abstract

l-myo-inositol-1-phosphate synthase (MIPS; EC: 5.5.1.4) activity has been detected and partially purified for the first time from human fetal liver. Crude homogenate from the fetal liver was subjected to streptomycin sulphate precipitation and 0–60 % ammonium sulphate fractionation followed by successive chromatography through DEAE cellulose and BioGel A 0.5-m columns. After the final chromatography, the enzyme was purified 51-fold and 3.46 % of MIPS could be recovered. The human fetal liver MIPS specifically utilised d-glucose-6-phosphte and NAD+ as its substrate and coenzyme, respectively. It shows pH optima between 7.0 and 7.5 while the temperature maximum was at 40 °C. The enzyme activity was remarkably stimulated by NH +4 , slightly stimulated by K+ and Ca2+ and highly inhibited by Zn2+, Cu2+ and Hg2+. The K m values of MIPS for d-glucose-6-phosphate and NAD+ were found to be as 1.15 and 0.12 mM respectively while the V max values were 280 nM and 252 nM for d-glucose-6-phosphate and NAD+ correspondingly. The apparent molecular weight of the native enzyme was determined to be 170 kDa.

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Abbreviations

G-6-P:

d-glucose-6-phosphate

MIPS:

l-myo-inositol-1-phosphate synthase

MIP:

l-myo-inositol-1-phosphate

ME:

2-mercaptoethanol

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Authors and Affiliations

  1. Biochemistry and Molecular Biology Laboratory, Darjeeling Government College, Darjeeling, West Bengal, India

    Dhani Raj Chhetri & Seema Gupta

  2. Biochemistry and Molecular Biology Laboratory, Burdwan University, Golapbag, Burdwan, West Bengal, India

    Asok Kumar Mukherjee

  3. Biochemistry Laboratory, Presidency University, Kolkata, West Bengal, India

    Jukta Adhikari

  4. Department of Ethnobotany and Social Medicine Studies, Sikkim University, Narsing-Jhelum Evaluation Block, 5th Mile-Tadong, Gangtok, 737102, Sikkim, India

    Dhani Raj Chhetri

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  1. Dhani Raj Chhetri
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  2. Seema Gupta
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  3. Asok Kumar Mukherjee
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  4. Jukta Adhikari
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Correspondence to Dhani Raj Chhetri.

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Chhetri, D.R., Gupta, S., Mukherjee, A.K. et al. l-myo-Inositol-1-Phosphate Synthase Expressed in Developing Organ: Isolation and Characterisation of the Enzyme from Human Fetal Liver. Appl Biochem Biotechnol 167, 2269–2282 (2012). https://doi.org/10.1007/s12010-012-9767-8

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