Expanding the Limits of Thermoacidophily in the Archaeon Sulfolobus solfataricus by Adaptive Evolution

Appl Environ Microbiol. 2015 Nov 20;82(3):857-67. doi: 10.1128/AEM.03225-15. Print 2016 Feb 1.

Abstract

Extremely thermoacidophilic Crenarchaeota belonging to the order Sulfolobales flourish in hot acidic habitats that are strongly oxidizing. The pH extremes of these habitats, however, often exceed the acid tolerance of type species and strains. Here, adaptive laboratory evolution was used over a 3-year period to test whether such organisms harbor additional thermoacidophilic capacity. Three distinct cell lines derived from a single type species were subjected to high-temperature serial passage while culture acidity was gradually increased. A 178-fold increase in thermoacidophily was achieved after 29 increments of shifted culture pH resulting in growth at pH 0.8 and 80°C. These strains were named super-acid-resistant Crenarchaeota (SARC). Mathematical modeling using growth parameters predicted the limits of acid resistance, while genome resequencing and transcriptome resequencing were conducted for insight into mechanisms responsible for the evolved trait. Among the mutations that were detected, a set of eight nonsynonymous changes may explain the heritability of increased acid resistance despite an unexpected lack of transposition. Four multigene components of the SARC transcriptome implicated oxidative stress as a primary challenge accompanying growth at acid extremes. These components included accelerated membrane biogenesis, induction of the mer operon, and an increased capacity for the generation of energy and reductant.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Adaptation, Physiological
  • Archaeal Proteins / genetics
  • Archaeal Proteins / metabolism
  • Biotechnology
  • Directed Molecular Evolution*
  • Genome, Bacterial
  • Hot Temperature*
  • Hydrogen-Ion Concentration
  • Models, Biological
  • Multigene Family
  • Mutation
  • Operon
  • Oxidation-Reduction
  • Oxidative Stress / genetics
  • Sequence Analysis, DNA
  • Sulfolobus solfataricus / genetics*
  • Sulfolobus solfataricus / growth & development
  • Sulfolobus solfataricus / physiology*
  • Time Factors
  • Transcriptome

Substances

  • Archaeal Proteins

Grants and funding

This work was supported by National Science Foundation grant MCB 1517408, the Nebraska Center for Energy Science Research, the UNL Cell Development Facility, and the Department of Energy Joint Genome Institute (DOE-JGI) under the community sequencing program (CSP Proposal 1218, projects 1019966, 1019969, and 1019972). The work conducted by the DOE-JGI, a DOE Office of Science User Facility, was supported by the Office of Science of the U.S. Department of Energy under contract no. DE-AC02-05CH11231.