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High-Level Production of Bacteriotoxic Phospholipase A1 in Bacterial Host Pseudomonas fluorescens via ABC Transporter-Mediated Secretion and Inducible Expression
- Source :
- Microorganisms, Vol 8, Iss 2, p 239 (2020), Microorganisms, Volume 8, Issue 2
- Publication Year :
- 2020
- Publisher :
- MDPI AG, 2020.
-
Abstract
- Bacterial phospholipase A1 (PLA1) is used in various industrial fields because it can catalyze the hydrolysis, esterification, and transesterification of phospholipids to their functional derivatives. It also has a role in the degumming process of crude plant oils. However, bacterial expression of the foreign PLA1-encoding gene was generally hampered because intracellularly expressed PLA1 is inherently toxic and damages the phospholipid membrane. In this study, we report that secretion-based production of recombinant PlaA, a bacterial PLA1 gene, or co-expression of PlaS, an accessory gene, minimizes this harmful effect. We were able to achieve high-level PlaA production via secretion-based protein production. Here, TliD/TliE/TliF, an ABC transporter complex of Pseudomonas fluorescens SIK-W1, was used to secrete recombinant proteins to the extracellular medium. In order to control the protein expression with induction, a new strain of P. fluorescens, which had the lac operon repressor gene lacI, was constructed and named ZYAI strain. The bacteriotoxic PlaA protein was successfully produced in a bacterial host, with help from ABC transporter-mediated secretion, induction-controlled protein expression, and fermentation. The final protein product is capable of degumming oil efficiently, signifying its application potential.
- Subjects :
- Microbiology (medical)
Repressor
lac operon
Pseudomonas fluorescens
ATP-binding cassette transporter
Lac repressor
Microbiology
Article
03 medical and health sciences
Phospholipase A1
Virology
phospholipase a1 (pla1)
protein secretion
Secretion
lcsh:QH301-705.5
030304 developmental biology
0303 health sciences
biology
030306 microbiology
Chemistry
pseudomonas fluorescens
biology.organism_classification
Secretory protein
Biochemistry
lcsh:Biology (General)
plaa, abc transporter
Subjects
Details
- Language :
- English
- ISSN :
- 20762607
- Volume :
- 8
- Issue :
- 2
- Database :
- OpenAIRE
- Journal :
- Microorganisms
- Accession number :
- edsair.doi.dedup.....2fb98ddf6a2a993dcd165baab28de2f4