Simplified high-throughput screening of AOX1-expressed laccase enzyme in Pichia pastoris

Current advances of Pichia pastoris as cell factories for production of recombinant proteins

“…The low efficiency of various modifications requires for rapid and functional perturbations. High throughput screening is an advanced and rapid laboratory technique that has been used in many microorganisms before ( Sjostrom et al, 2014 ; Kenzom et al, 2015 ; Clare et al, 2019 ). The identification of desired results combined with the appliance of reverse engineering can enable the attainment of strains carrying specific mutations associated with the desired phenotype ( Wang G. K. et al, 2017 ).…”

“…The identification of desired results combined with the appliance of reverse engineering can enable the attainment of strains carrying specific mutations associated with the desired phenotype ( Wang G. K. et al, 2017 ). High-throughput screening method can be simplified by carrying out the growth phase and induction phase together by inoculation of P. pastoris clones directly into methanol-containing medium ( Kenzom et al, 2015 ). In addition, high-throughput screening help understand the impact of genetic engineering such as ectopic integration.…”

“…Compared with traditional microplate screening, the reagent cost was reduced by millions of times ( Lu et al, 2019 ). With atmospheric and room temperature plasma (ARTP) mutagenesis and droplet-based microfluidic high-throughput screening, it is possible to explore the potential of protein production by employing iterative rounds of genome-wide diversity generation ( Kenzom et al, 2015 ; Ma et al, 2019 ; Yuan et al, 2022 ). For example, compared to the beginning strain, the best mutant strain showed a twofold increase in cellulase activity in one research ( Yuan et al, 2022 ).…”