Construction of a Pseudozyma antarctica strain without foreign DNA sequences (self-cloning strain) for high yield production of a biodegradable plastic-degrading enzyme ...

• Main Authors: Sameshima-Yamashita, Yuka, Watanabe, Takashi, Tanaka, Takumi, Tsuboi, Shun, Yarimizu, Tohru, Morita, Tomotake, Koike, Hideaki, Suzuki, Ken, Kitamoto, Hiroko
• Format: Text
• Language: unknown
• Published: Taylor & Francis 2019
• Subjects: Biochemistry; Space Science; Microbiology; FOS Biological sciences; Genetics; Molecular Biology; Biotechnology; Ecology; Biological Sciences not elsewhere classified; Developmental Biology; Science Policy; Infectious Diseases; FOS Health sciences; Antarc*; Antarctica
• Online Access: https://dx.doi.org/10.6084/m9.figshare.7666973.v1; https://tandf.figshare.com/articles/journal_contribution/Construction_of_a_i_Pseudozyma_antarctica_i_strain_without_foreign_DNA_sequences_self-cloning_strain_for_high_yield_production_of_a_biodegradable_plastic-degrading_enzyme/7666973/1

The basidiomycetous yeast Pseudozyma antarctica GB-4(0) esterase (PaE) is a promising candidate for accelerating degradation of used biodegradable plastics (BPs). To increase safety and reduce costs associated with the use of PaE, we constructed a self-cloning strain with high-PaE productivity. A Lys12 gene (Pa LYS12 )-deleted lysine auxotroph strain GB4-(0)-L1 was obtained from GB-4(0) by ultraviolet mutagenesis and nystatin enrichment. Subsequently, the PaE gene (Pa CLE1 ) expression cassette consisting of GB-4(0)-derived Pa CLE1 , under the control of a xylose-inducible xylanase promoter with Pa LYS12 , was randomly introduced into the GB4-(0)-L1 genome. A PaE high-producing strain, PGB474, was selected from among the transformants by high throughput double-screening based on its ability to degrade emulsified polybutylene succinate- co -adipate. Quantitative PCR revealed that four copies of the PaE gene expression cassette were introduced into the PGB474 genome. PGB474 produced 2.0 g/L of PaE by ...