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Production of Squalene in Bacillus subtilis by Squalene Synthase Screening and Metabolic Engineering

Authors
  • Song, Yafeng1
  • Guan, Zheng1
  • van Merkerk, Ronald1
  • Pramastya, Hegar1, 2
  • Abdallah, Ingy I.1, 3
  • Setroikromo, Rita1
  • Quax, Wim J.1
  • 1 University of Groningen, The Netherlands
  • 2 Institut Teknologi Bandung, Indonesia , (Indonesia)
  • 3 Alexandria University, Egypt , (Egypt)
Type
Published Article
Journal
Journal of Agricultural and Food Chemistry
Publisher
American Chemical Society
Publication Date
Mar 25, 2020
Volume
68
Issue
15
Pages
4447–4455
Identifiers
DOI: 10.1021/acs.jafc.0c00375
PMID: 32208656
PMCID: PMC7168599
Source
PubMed Central
Keywords
License
Unknown

Abstract

Squalene synthase (SQS) catalyzes the conversion of two farnesyl pyrophosphates to squalene, an important intermediate in between isoprene and valuable triterpenoids. In this study, we have constructed a novel biosynthesis pathway for squalene in Bacillus subtilis and performed metabolic engineering aiming at facilitating further exploitation and production of squalene-derived triterpenoids. Therefore, systematic studies and analysis were performed including selection of multiple SQS candidates from various organisms, comparison of expression vectors, optimization of cultivation temperatures, and examination of rate-limiting factors within the synthetic pathway. We were, for the first time, able to obtain squalene synthesis in B. subtilis . Furthermore, we achieved a 29-fold increase of squalene yield (0.26–7.5 mg/L) by expressing SQS from Bacillus megaterium and eliminating bottlenecks within the upstream methylerythritol-phosphate pathway. Moreover, our findings showed that also ispA could positively affect the production of squalene.

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