Multidimensional Engineering of Cyberlindnera jadinii for Efficient Invertase Production

Area of Science:

• Biotechnology
• Microbial Engineering
• Industrial Microbiology

Background:

• Cyberlindnera jadinii yeast is utilized in food and feed industries.
• Limited protein secretion capacity hinders its use as a host for protein production.

Purpose of the Study:

• To engineer Cyberlindnera jadinii for enhanced extracellular protein production.
• To overcome limitations in protein secretion using genetic and cell wall engineering.

Main Methods:

• Multidimensional engineering of a tetraploid wild-type strain (NBRC0988).
• Multicopy integration of gene expression cassettes using a landing pad strategy.
• Transcriptional enhancement via promoter/terminator optimization and chitin-based cell wall remodeling.

Main Results:

• Engineered strain demonstrated significantly enhanced protein secretion in a 5-L bioreactor.
• Achieved an extracellular protein yield of 3456 ± 81 mg/L after 168 hours of cultivation.
• Attained invertase specific activity of 789.3 ± 52.3 U/mg, a record for yeast-based invertase production.

Conclusions:

• Cyberlindnera jadinii can be engineered into a high-performance platform for recombinant protein production.
• The developed multidimensional engineering strategy effectively enhances protein secretion.
• This study establishes a new benchmark for yeast-based extracellular protein yields.