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Cited by 118Cassava research and cyanidePlant Toxicity and Pharmacological PropertiesEnzyme Structure and Function

Evolution of heteromeric nitrilase complexes in Poaceae with new functions in nitrile metabolism

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Jenrich, Roland, Trompetter, Inga, Bak, Søren, Olsen, Carl Erik, Møller, Birger Lindberg, Piotrowski, Markus·2007·Proceedings of the National Academy of Sciences
DOI 10.1073/pnas.0709315104PMC PMC2141865
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Abstract

Members of the nitrilase 4 (NIT4) family of higher plants catalyze the conversion of β-cyanoalanine to aspartic acid and asparagine, a key step in cyanide detoxification. Grasses (Poaceae) possess two different NIT4 homologs (NIT4A and NIT4B), but none of the recombinant Poaceae enzymes analyzed showed activity with β-cyanoalanine, whereas protein extracts of the same plants clearly posses this activity. Sorghum bicolor contains three NIT4 isoforms Sb NIT4A, Sb NIT4B1, and Sb NIT4B2. Individually, each isoform does not possess enzymatic activity whereas the heteromeric complexes Sb NIT4A/B1 and Sb NIT4A/B2 hydrolyze β-cyanoalanine with high activity. In addition, the Sb NIT4A/B2 complex accepts additional substrates, the best being 4-hydroxyphenylacetonitrile. Corresponding NIT4A and NIT4B isoforms from other Poaceae species can functionally complement the sorghum isoforms in these complexes. Site-specific mutagenesis of the active site cysteine residue demonstrates that hydrolysis of β-cyanoalanine is catalyzed by the NIT4A isoform in both complexes whereas hydrolysis of 4-hydroxyphenylacetonitrile occurs at the NIT4B2 isoform. 4-Hydroxyphenylacetonitrile was shown to be an in vitro breakdown product of the cyanogenic glycoside dhurrin, a main constituent in S. bicolor . The results indicate that the Sb NIT4A/B2 heterocomplex plays a key role in an endogenous turnover of dhurrin proceeding via 4-hydroxyphenylacetonitrile and thereby avoiding release of toxic hydrogen cyanide. The operation of this pathway would enable plants to use cyanogenic glycosides as transportable and remobilizable nitrogenous storage compounds. Through combinatorial biochemistry and neofunctionalizations, the small family of nitrilases has gained diverse biological functions in nitrile metabolism.

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  • Two sorghum nitrilases are capable of producing Asn from beta-cyano-Ala.

    “Indeed, two sorghum (Sorghum bicolor) nitrilases are capable of producing Asn from b-cyano-Ala (Jenrich et al., 2007).”

    From Plant Secondary Metabolites as Defenses, Regulators, and Primary Metabolites: The Blurred Functional Trichotomy (2020) · cites this paper as Jenrich et al., 2007

    The Abstract and Results report that two sorghum heteromeric complexes, SbNIT4A/B1 and SbNIT4A/B2, metabolize β-cyanoalanine; the paper describes asparagine as a product of this NIT4 reaction. But the claim omits the essential qualification that the sorghum nitrilase isoforms are inactive individually: the activity belongs to the two complexes, not to two standalone enzymes. (LLM, conf=0.86)

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Cited by

Sorted by citations

Authors
Jenrich, R., Trompetter, I., Bak, S., Olsen, C. E., Møller, B. L., & Piotrowski, M.
Affiliations
  • Ruhr University Bochum (DE)
  • University of Copenhagen (DK)
ORCID
Venue
Proceedings of the National Academy of Sciences
Publisher
National Academy of Sciences
Published
2007-11-20
Volume
104
Issue
47
Pages
18848-18853
ISSN
  • 0027-8424 (ISSN-L)
  • 1091-6490
DOI
10.1073/pnas.0709315104
PMC
PMC2141865
OpenAlex
W2147203026
Cited by
118
References
1
Citations by year
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FWCI
11.75
Type
journal article
Language
en
Open access
Locations
  • Proceedings of the National Academy of Sciences (publishedVersion)
  • Europe PMC (PubMed Central) (publishedVersion)
  • PubMed Central (publishedVersion)
  • PubMed (publishedVersion)
  • Research at the University of Copenhagen (University of Copenhagen) (submittedVersion)
Topics
  • Life Sciences › Agricultural and Biological Sciences › Plant Science › Cassava research and cyanide
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Plant Toxicity and Pharmacological Properties
  • Physical Sciences › Materials Science › Materials Chemistry › Enzyme Structure and Function
Keywords
Poaceae, sorghum, cyanide detoxification, dhurrin, cyanogenic glycosides, nitrile metabolism, neofunctionalization, active site cysteine, nitrogen remobilization
MeSH terms
Alanine, Aminohydrolases, Binding Sites, Cyanides, Poaceae, Kinetics, Nitriles, Phylogeny, Protein Binding, Recombinant Proteins, Substrate Specificity, Molecular Structure, Evolution, Molecular

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