Discovery of cyanophycin dipeptide hydrolase enzymes suggests widespread utility of the natural biopolymer cyanophycin.

Publication Type:

Journal Article

Source:

Proc Natl Acad Sci U S A, Volume 120, Issue 8, p.e2216547120 (2023)

Keywords:

Bacteria, Bacterial Proteins, Biopolymers, Dipeptides, Nitrogen, Polymers

Abstract:

<p>Cyanophycin is a bacterial polymer mainly used for nitrogen storage. It is composed of a peptide backbone of L-aspartate residues with L-arginines attached to their side chains through isopeptide bonds. Cyanophycin is degraded in two steps: Cyanophycinase cleaves the polymer into β-Asp-Arg dipeptides, which are hydrolyzed into free Asp and Arg by enzymes possessing isoaspartyl dipeptide hydrolase activity. Two unrelated enzymes with this activity,&nbsp;isoaspartyl dipeptidase (IadA) and isoaspartyl aminopeptidase (IaaA) have been shown to degrade β-Asp-Arg dipeptides, but bacteria which encode cyanophycin-metabolizing genes can lack and genes. In this study, we investigate a previously uncharacterized enzyme whose gene can cluster with cyanophycin-metabolizing genes. This enzyme, which we name cyanophycin dipeptide hydrolase (CphZ), is specific for dipeptides derived from cyanophycin degradation. Accordingly, a co-complex structure of CphZ and β-Asp-Arg shows that CphZ, unlike IadA or IaaA, recognizes all portions of its β-Asp-Arg substrate. Bioinformatic analyses showed that CphZ is found in very many proteobacteria and is homologous to an uncharacterized protein encoded in the &quot;arginine/ornithine transport&quot; (aot) operon of many pseudomonas species, including . In&nbsp;vitro assays show that AotO is indeed a CphZ, and in cellulo growth experiments show that this enzyme and the operon allow to take up and use β-Asp-Arg as a sole carbon and nitrogen source. Together the results establish the novel, highly specific enzyme subfamily of CphZs, suggesting that cyanophycin is potentially used by a much wider range of bacteria than previously appreciated.</p>

PDB: 
8EIN (AbCphZ) and 8EIP (AbCphZ_E251A + β-Asp-Arg).
Detector: 
EIGER2
Beamline: 
24-ID-C