{"id":13470,"date":"2024-01-18T10:09:34","date_gmt":"2024-01-18T10:09:34","guid":{"rendered":"https:\/\/www.nanion.de\/?post_type=resource_library&#038;p=13470"},"modified":"2024-01-18T10:10:14","modified_gmt":"2024-01-18T10:10:14","slug":"2024-a-cost-effective-pichia-pastoris-cell-free-system-driven-by-glycolytic-intermediates-enables-the-production-of-complex-eukaryotic-proteins","status":"publish","type":"resource_library","link":"https:\/\/nanion.com\/ja\/resource_library\/2024-a-cost-effective-pichia-pastoris-cell-free-system-driven-by-glycolytic-intermediates-enables-the-production-of-complex-eukaryotic-proteins\/","title":{"rendered":"2024 &#8211; A Cost-Effective Pichia pastoris Cell-Free System Driven by Glycolytic Intermediates Enables the Production of Complex Eukaryotic Proteins"},"content":{"rendered":"<p>Cell-free systems are particularly attractive for screening applications and the production of difficult-to-express proteins. However, the production of cell lysates is difficult to implement on a larger scale due to large time requirements, cultivation costs, and the supplementation of cell-free reactions with energy regeneration systems. Consequently, the methylotrophic yeast <em>Pichia pastoris<\/em>, which is widely used in recombinant protein production, was utilized in the present study to realize cell-free synthesis in a cost-effective manner. Sensitive disruption conditions were evaluated, and appropriate signal sequences for translocation into ER vesicles were identified. An alternative energy regeneration system based on fructose-1,6-bisphosphate was developed and a ~2-fold increase in protein production was observed. Using a statistical experiment design, the optimal composition of the cell-free reaction milieu was determined. Moreover, functional ion channels could be produced, and a G-protein-coupled receptor was site-specifically modified using the novel cell-free system. Finally, the established <em>P. pastoris<\/em> cell-free protein production system can economically produce complex proteins for biotechnological applications in a short time.<\/p>\n","protected":false},"featured_media":0,"template":"","products_tags":[98],"resource_type":[75],"application_areas":[50,53,99],"therapeutic_areas":[143],"target_categories":[341],"target_families":[349],"targets":[310],"resource_keyword":[],"class_list":["post-13470","resource_library","type-resource_library","status-publish","hentry","products_tags-orbit-16-tc","resource_type-publication","application_areas-ion-channels","application_areas-assay-development-and-validation","application_areas-pores-and-toxins","therapeutic_areas-assay-development-and-validation","target_categories-pores-and-toxins","target_families-prokaryotic-pores","targets-kcsa"],"acf":[],"_links":{"self":[{"href":"https:\/\/nanion.com\/ja\/wp-json\/wp\/v2\/resource_library\/13470","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/nanion.com\/ja\/wp-json\/wp\/v2\/resource_library"}],"about":[{"href":"https:\/\/nanion.com\/ja\/wp-json\/wp\/v2\/types\/resource_library"}],"wp:attachment":[{"href":"https:\/\/nanion.com\/ja\/wp-json\/wp\/v2\/media?parent=13470"}],"wp:term":[{"taxonomy":"products_tags","embeddable":true,"href":"https:\/\/nanion.com\/ja\/wp-json\/wp\/v2\/products_tags?post=13470"},{"taxonomy":"resource_type","embeddable":true,"href":"https:\/\/nanion.com\/ja\/wp-json\/wp\/v2\/resource_type?post=13470"},{"taxonomy":"application_areas","embeddable":true,"href":"https:\/\/nanion.com\/ja\/wp-json\/wp\/v2\/application_areas?post=13470"},{"taxonomy":"therapeutic_areas","embeddable":true,"href":"https:\/\/nanion.com\/ja\/wp-json\/wp\/v2\/therapeutic_areas?post=13470"},{"taxonomy":"target_categories","embeddable":true,"href":"https:\/\/nanion.com\/ja\/wp-json\/wp\/v2\/target_categories?post=13470"},{"taxonomy":"target_families","embeddable":true,"href":"https:\/\/nanion.com\/ja\/wp-json\/wp\/v2\/target_families?post=13470"},{"taxonomy":"targets","embeddable":true,"href":"https:\/\/nanion.com\/ja\/wp-json\/wp\/v2\/targets?post=13470"},{"taxonomy":"resource_keyword","embeddable":true,"href":"https:\/\/nanion.com\/ja\/wp-json\/wp\/v2\/resource_keyword?post=13470"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}