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A method for the efficient adenosylation of corrinoids.

Flavia G Costa1, Elizabeth A Villa1, Jorge C Escalante-Semerena1

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|May 19, 2022
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Summary

This study details a new method for enzymatically synthesizing adenosylcobamides (AdoCbas), essential coenzymes. The described technique also allows for the quantification of these vital organometallic compounds.

Keywords:
ACATB(12) adenosylationCobamide adenosyltransferaseCoenzyme B(12) biosynthesisCorrinoid adenosylationCorrinoid adenosyltransferaseVitamin B(12) to coenzyme B(12) conversion

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Area of Science:

  • Biochemistry
  • Enzymology
  • Organometallic Chemistry

Background:

  • Adenosylcobamides (AdoCbas) are crucial coenzymes across all life domains, facilitating complex chemical reactions via a unique cobalt-carbon organometallic bond.
  • This Co-C bond is cleaved during enzymatic catalysis to generate an adenosyl radical, essential for intramolecular rearrangements.
  • ATP:Co(I)rrinoid adenosyltransferases (ACATs) catalyze AdoCba synthesis through a two-step process involving corrinoid planarization, reduction, and adenosyl group transfer from ATP.

Purpose of the Study:

  • To describe an enzymatic method for the synthesis of specific adenosylcobamides (AdoCbas).
  • To provide a means for the quantification of synthesized AdoCbas.
  • To address the challenge of maintaining anoxic conditions for in vitro synthesis.

Main Methods:

  • Enzymatic synthesis of AdoCbas using ATP:Co(I)rrinoid adenosyltransferases (ACATs).
  • Strict maintenance of anoxic conditions to prevent oxidation of cobalt ions (Co(II) and Co(I)).
  • Development of a quantification method for the synthesized AdoCbas.

Main Results:

  • Successful enzymatic synthesis of specific adenosylcobamides (AdoCbas) was achieved.
  • A method for quantifying these synthesized AdoCbas was established.
  • The described method facilitates the production of these essential coenzymes under controlled conditions.

Conclusions:

  • The developed method enables the efficient enzymatic synthesis and quantification of AdoCbas.
  • This work provides a valuable tool for researchers studying AdoCba-dependent enzymes and reactions.
  • The described protocol aids in understanding the synthesis and function of these vital biological molecules.