AI Article Synopsis
- Methanobactin (Mbn) is a natural peptide that effectively binds and chelates copper ions, produced by the bacteria Methylosinus trichosporium OB3b through a complex biosynthetic process involving various enzymes.
- The core enzyme complex MbnBC modifies a precursor peptide (MbnA) to form Mbn, with additional modifications by the aminotransferase MbnN after a cleavage step.
- The text outlines detailed methods for expressing and purifying the necessary enzymes and for studying the structural and functional aspects of MbnBC, with broader implications for synthesizing Mbn-like compounds for potential therapeutic uses.
Article Abstract
Methanobactin (Mbn) is a ribosomally synthesized and post-translationally modified peptide (RiPP) natural product that binds Cu(I) with high affinity. The copper-chelating thioamide/oxazolone groups in Mbn are installed on the precursor peptide MbnA by the core enzyme complex, MbnBC, which includes the multinuclear non-heme iron-dependent oxidase (MNIO) MbnB and its RiPP recognition element-containing partner protein MbnC. For the extensively characterized Mbn biosynthetic gene cluster (BGC) from the methanotroph Methylosinus trichosporium OB3b, the tailoring aminotransferase MbnN further modifies MbnA after leader sequence cleavage by an unknown mechanism. Here we detail methods to express and purify M. trichosporium OB3b MbnBC and MbnN along with protocols for assessing MbnA modification by MbnBC and MbnN aminotransferase activity. In addition, we describe crystallization and structure determination of MbnBC. These procedures can be adapted for other MNIOs and partner proteins encoded in Mbn and Mbn-like BGCs. Furthermore, these methods provide a first step toward in vitro biosynthesis of Mbns and related natural products as potential therapeutics.
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| http://www.ncbi.nlm.nih.gov/pmc/articles/PMC11622243 | PMC |
| http://dx.doi.org/10.1016/bs.mie.2024.06.011 | DOI Listing |
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Article Synopsis
- Methanobactin (Mbn) is a natural peptide that effectively binds and chelates copper ions, produced by the bacteria Methylosinus trichosporium OB3b through a complex biosynthetic process involving various enzymes.
- The core enzyme complex MbnBC modifies a precursor peptide (MbnA) to form Mbn, with additional modifications by the aminotransferase MbnN after a cleavage step.
- The text outlines detailed methods for expressing and purifying the necessary enzymes and for studying the structural and functional aspects of MbnBC, with broader implications for synthesizing Mbn-like compounds for potential therapeutic uses.
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