Metabolic engineering of probiotic Saccharomyces boulardii

Jing Jing Liu, In Iok Kong, Guo Chang Zhang, Lahiru N. Jayakody, Heejin Kim, Peng Fei Xia, Suryang Kwak, Bong Hyun Sung, Jung Hoon Sohn, Hanna E. Walukiewicz, Christopher V. Rao, Yong Su Jin

Research output: Contribution to journalArticle

Abstract

Saccharomyces boulardii is a probiotic yeast that has been used for promoting gut health as well as preventing diarrheal diseases. This yeast not only exhibits beneficial phenotypes for gut health but also can stay longer in the gut than Saccharomyces cerevisiae. Therefore, S. boulardii is an attractive host for metabolic engineering to produce biomolecules of interest in the gut. However, the lack of auxotrophic strains with defined genetic backgrounds has hampered the use of this strain for metabolic engineering. Here, we report the development of well-defined auxotrophic mutants (leu2, ura3, his3, and trp1) through clustered regularly interspaced short palindromic repeat (CRISPR)-Cas9-based genome editing. The resulting auxotrophic mutants can be used as a host for introducing various genetic perturbations, such as overexpression or deletion of a target gene, using existing genetic tools for S. cerevisiae. We demonstrated the overexpression of a heterologous gene (lacZ), the correct localization of a target protein (red fluorescent protein) into mitochondria by using a protein localization signal, and the introduction of a heterologous metabolic pathway (xylose-assimilating pathway) in the genome of S. boulardii. We further demonstrated that human lysozyme, which is beneficial for human gut health, could be secreted by S. boulardii. Our results suggest that more sophisticated genetic perturbations to improve S. boulardii can be performed without using a drug resistance marker, which is a prerequisite for in vivo applications using engineered S. boulardii.

Original languageEnglish (US)
Pages (from-to)2280-2287
Number of pages8
JournalApplied and environmental microbiology
Volume82
Issue number8
DOIs
StatePublished - Apr 1 2016

Fingerprint

Saccharomyces cerevisiae var. cerevisiae
Metabolic Engineering
metabolic engineering
probiotics
Probiotics
auxotrophs
digestive system
engineering
protein
yeast
genome
perturbation
diarrheal disease
drug resistance
Saccharomyces cerevisiae
gene
Health
mitochondrion
Clustered Regularly Interspaced Short Palindromic Repeats
Yeasts

ASJC Scopus subject areas

  • Biotechnology
  • Food Science
  • Applied Microbiology and Biotechnology
  • Ecology

Cite this

Liu, J. J., Kong, I. I., Zhang, G. C., Jayakody, L. N., Kim, H., Xia, P. F., ... Jin, Y. S. (2016). Metabolic engineering of probiotic Saccharomyces boulardii. Applied and environmental microbiology, 82(8), 2280-2287. https://doi.org/10.1128/AEM.00057-16

Metabolic engineering of probiotic Saccharomyces boulardii. / Liu, Jing Jing; Kong, In Iok; Zhang, Guo Chang; Jayakody, Lahiru N.; Kim, Heejin; Xia, Peng Fei; Kwak, Suryang; Sung, Bong Hyun; Sohn, Jung Hoon; Walukiewicz, Hanna E.; Rao, Christopher V.; Jin, Yong Su.

In: Applied and environmental microbiology, Vol. 82, No. 8, 01.04.2016, p. 2280-2287.

Research output: Contribution to journalArticle

Liu, JJ, Kong, II, Zhang, GC, Jayakody, LN, Kim, H, Xia, PF, Kwak, S, Sung, BH, Sohn, JH, Walukiewicz, HE, Rao, CV & Jin, YS 2016, 'Metabolic engineering of probiotic Saccharomyces boulardii', Applied and environmental microbiology, vol. 82, no. 8, pp. 2280-2287. https://doi.org/10.1128/AEM.00057-16
Liu JJ, Kong II, Zhang GC, Jayakody LN, Kim H, Xia PF et al. Metabolic engineering of probiotic Saccharomyces boulardii. Applied and environmental microbiology. 2016 Apr 1;82(8):2280-2287. https://doi.org/10.1128/AEM.00057-16
Liu, Jing Jing ; Kong, In Iok ; Zhang, Guo Chang ; Jayakody, Lahiru N. ; Kim, Heejin ; Xia, Peng Fei ; Kwak, Suryang ; Sung, Bong Hyun ; Sohn, Jung Hoon ; Walukiewicz, Hanna E. ; Rao, Christopher V. ; Jin, Yong Su. / Metabolic engineering of probiotic Saccharomyces boulardii. In: Applied and environmental microbiology. 2016 ; Vol. 82, No. 8. pp. 2280-2287.
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