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The application relates to the field of biotechnology, and particularly discloses a mutant strain Cbg61-T37, a co-expression strain Cbg61-T37-MPR1 and a construction method and application thereof. The mutant strain Cbg61-T37 is preserved in the China General Microbiological Culture Collection Center on August 18, 2025, has a preservation number of CGMCC No. 35649, and is classified and named as Pichia pastoris. Pichia pastoris The mutant strain Cbg61-T37 provided by the application has an enzyme hydrolysis activity of 62.9 U / mg, which is increased by 43.8% compared to the original strain. Therefore, the mutant strain Cbg61-T37 is more conducive to catalysis and synthesis of the rare ginsenoside CK.