Discovery of Novel Pseudomonas putida Flavin-Binding Fluorescent Protein Variants with Significantly Improved Quantum Yield

Sanghwan Ko, Sanghwan Ko, Hyunwoo Jeon, Sanghan Yoon, Munsu Kyung, Hyungdon Yun, Jung Hyun Na, Sang Taek Jung

Research output: Contribution to journalArticlepeer-review

1 Citation (Scopus)

Abstract

Oxygen-independent, flavin-binding fluorescent proteins (FbFPs) are emerging as alternatives to green fluorescent protein (GFP), which has limited applicability in studying anaerobic microorganisms, such as human gastrointestinal bacteria, which grow in oxygen-deficient environments. However, the utility of these FbFPs has been compromised because of their poor fluorescence emission. To overcome this limitation, we have employed a high-throughput library screening strategy and engineered an FbFP derived from Pseudomonas putida (SB2) for enhanced quantum yield. Of the resulting SB2 variants, KOFP-7 exhibited a significantly improved quantum yield (0.61) compared to other reported engineered FbFPs, which was even higher than that of enhanced GFP (EGFP, 0.60), with significantly enhanced tolerance against a strong reducing agent.

Original languageEnglish
Pages (from-to)5873-5879
Number of pages7
JournalJournal of agricultural and food chemistry
Volume68
Issue number21
DOIs
Publication statusPublished - 2020 May 27

Keywords

  • flavin-binding fluorescent protein
  • fluorescence intensity
  • quantum yield

ASJC Scopus subject areas

  • Chemistry(all)
  • Agricultural and Biological Sciences(all)

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