Skip to main content
Journal cover image

Identification of cyclosporin C from Amphichorda felina using a Cryptococcus neoformans differential temperature sensitivity assay.

Publication ,  Journal Article
Xu, L; Li, Y; Biggins, JB; Bowman, BR; Verdine, GL; Gloer, JB; Alspaugh, JA; Bills, GF
Published in: Appl Microbiol Biotechnol
March 2018

We used a temperature differential assay with the opportunistic fungal pathogen Cryptococcus neoformans as a simple screening platform to detect small molecules with antifungal activity in natural product extracts. By screening of a collection extracts from two different strains of the coprophilous fungus, Amphichorda felina, we detected strong, temperature-dependent antifungal activity using a two-plate agar zone of inhibition assay at 25 and 37 °C. Bioassay-guided fractionation of the crude extract followed by liquid chromatography-mass spectrometry (LC-MS) and nuclear magnetic resonance spectroscopy (NMR) identified cyclosporin C (CsC) as the main component of the crude extract responsible for growth inhibition of C. neoformans at 37 °C. The presence of CsC was confirmed by comparison with a commercial standard. We sequenced the genome of A. felina to identify and annotate the CsC biosynthetic gene cluster. The only previously characterized gene cluster for the biosynthesis of similar compounds is that of the related immunosuppressant drug cyclosporine A (CsA). The CsA and CsC gene clusters share a high degree of synteny and sequence similarity. Amino acid changes in the adenylation domain of the CsC nonribosomal peptide synthase's sixth module may be responsible for the substitution of L-threonine compared to L-α-aminobutyric acid in the CsA peptide core. This screening strategy promises to yield additional antifungal natural products with a focused spectrum of antimicrobial activity.

Duke Scholars

Altmetric Attention Stats
Dimensions Citation Stats

Published In

Appl Microbiol Biotechnol

DOI

EISSN

1432-0614

Publication Date

March 2018

Volume

102

Issue

5

Start / End Page

2337 / 2350

Location

Germany

Related Subject Headings

  • Temperature
  • Hypocreales
  • Cyclosporins
  • Cryptococcus neoformans
  • Biotechnology
  • Antifungal Agents
 

Citation

APA
Chicago
ICMJE
MLA
NLM
Xu, L., Li, Y., Biggins, J. B., Bowman, B. R., Verdine, G. L., Gloer, J. B., … Bills, G. F. (2018). Identification of cyclosporin C from Amphichorda felina using a Cryptococcus neoformans differential temperature sensitivity assay. Appl Microbiol Biotechnol, 102(5), 2337–2350. https://doi.org/10.1007/s00253-018-8792-0
Xu, Lijian, Yan Li, John B. Biggins, Brian R. Bowman, Gregory L. Verdine, James B. Gloer, J Andrew Alspaugh, and Gerald F. Bills. “Identification of cyclosporin C from Amphichorda felina using a Cryptococcus neoformans differential temperature sensitivity assay.Appl Microbiol Biotechnol 102, no. 5 (March 2018): 2337–50. https://doi.org/10.1007/s00253-018-8792-0.
Xu L, Li Y, Biggins JB, Bowman BR, Verdine GL, Gloer JB, et al. Identification of cyclosporin C from Amphichorda felina using a Cryptococcus neoformans differential temperature sensitivity assay. Appl Microbiol Biotechnol. 2018 Mar;102(5):2337–50.
Xu, Lijian, et al. “Identification of cyclosporin C from Amphichorda felina using a Cryptococcus neoformans differential temperature sensitivity assay.Appl Microbiol Biotechnol, vol. 102, no. 5, Mar. 2018, pp. 2337–50. Pubmed, doi:10.1007/s00253-018-8792-0.
Xu L, Li Y, Biggins JB, Bowman BR, Verdine GL, Gloer JB, Alspaugh JA, Bills GF. Identification of cyclosporin C from Amphichorda felina using a Cryptococcus neoformans differential temperature sensitivity assay. Appl Microbiol Biotechnol. 2018 Mar;102(5):2337–2350.
Journal cover image

Published In

Appl Microbiol Biotechnol

DOI

EISSN

1432-0614

Publication Date

March 2018

Volume

102

Issue

5

Start / End Page

2337 / 2350

Location

Germany

Related Subject Headings

  • Temperature
  • Hypocreales
  • Cyclosporins
  • Cryptococcus neoformans
  • Biotechnology
  • Antifungal Agents