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Production of Synthetic Methionine-Free and Synthetic Methionine-Limited Alpha Casein: Protein Foodstuff for Patients with Homocystinuria due to Cystathionine Beta-Synthase Deficiency

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Abstract

The aim of this study was to evaluate the possibility that synthetic forms of methionine-free alpha-casein and methionine-limited alpha casein could be produced by recombinant means to form the basis for developing an industrial-scale process for the provision of a foodstuff suitable for patients with homocystinuria due to cystathionine beta-synthase (CBS) deficiency. As a first step, two forms of alpha casein gene, encoding methionine-free alpha casein (Fcas) or a methionine-limited alpha casein (Mcas), were synthesised and expressed in Escherichia coli. Using the overexpression vector pET28a, both genes were highly expressed in E. coli in soluble form as well as in inclusion bodies. The two recombinant proteins were purified by the one step methods using the fused His-tag and the Ni2+column and validated by Western blot analysis. This work paves the way for industrial-scale production of proteins suitable for patients with homocystinuria due to CBS deficiency.

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Abbreviations

SDS–PAGE:

Sodium dodecyl sulfate–polyacrylamide gel electrophoresis

IPTG:

Isopropyl-β-d-thiogalactopyranoside

CBS:

Cystathionine beta-synthase

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Acknowledgments

The authors would like to thank Shafalah Medical Genetic Centre (SMGC), Doha, Qatar for performing all the DNA sequencing and for other technical support. The author would like to thank Yaseen I. Gad for his help and support. This publication was made possible by a grant from the Qatar National Research Fund under its Undergraduate Research Experience Program. Its contents are solely the responsibility of the authors and do not necessarily represent the official views of the Qatar National Research Fund.

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Authors and Affiliations

  1. College of Arts and Sciences, Qatar University, P.O. Box 2713, Doha, Qatar

    Sayed K. Goda, Yasmin W. Abu Aqel & Mai R. Al-Aswad

  2. Chemistry Department, Faculty of Science, Cairo University, Cairo, Egypt

    Fatma A. B. Rashedy & Amr S. Mohamed

Authors
  1. Sayed K. Goda
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  2. Yasmin W. Abu Aqel
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  3. Mai R. Al-Aswad
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  4. Fatma A. B. Rashedy
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  5. Amr S. Mohamed
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Correspondence to Sayed K. Goda.

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Goda, S.K., Abu Aqel, Y.W., Al-Aswad, M.R. et al. Production of Synthetic Methionine-Free and Synthetic Methionine-Limited Alpha Casein: Protein Foodstuff for Patients with Homocystinuria due to Cystathionine Beta-Synthase Deficiency. Protein J 29, 44–49 (2010). https://doi.org/10.1007/s10930-009-9219-7

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  • DOI: https://doi.org/10.1007/s10930-009-9219-7

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