{"id":4720,"date":"2025-08-28T16:16:09","date_gmt":"2025-08-28T07:16:09","guid":{"rendered":"https:\/\/bionauts.jp\/?p=4720"},"modified":"2025-09-08T21:46:58","modified_gmt":"2025-09-08T12:46:58","slug":"expression-vector-for-streptomyces-spp","status":"publish","type":"post","link":"https:\/\/bionauts.jp\/?p=4720","title":{"rendered":"Expression Vector for Streptomyces spp."},"content":{"rendered":"<h3>Features and Vector Design<\/h3>\n<ul>\n<li>Designed as a shuttle vector replicable in <em> coli<\/em>: The plasmid can be constructed in E. coli and subsequently transferred to <em>Streptomyces spp.<\/em><\/li>\n<li>Inducer-free high-level expression: Transcription is activated by the strong promoter system derived from the <em>Rhodococcus<\/em> nitrile hydratase gene cluster, including the transcriptional regulators NhhC and NhhD.<\/li>\n<li>Optimized Tat secretion pathway: Efficient extracellular secretion of target proteins is achieved using Tat signal peptides derived from <em>Streptomyces avermitilis<\/em>.<\/li>\n<\/ul>\n<table style=\"border-collapse: collapse; width: 56.6667%; height: 131px;\">\n<tbody>\n<tr>\n<td style=\"width: 100%;\"><a href=\"http:\/\/bionauts.jp\/wp-content\/uploads\/2025\/08\/WL-05177.png\"><img decoding=\"async\" loading=\"lazy\" class=\"alignleft size-full wp-image-4721\" src=\"http:\/\/bionauts.jp\/wp-content\/uploads\/2025\/08\/WL-05177.png\" alt=\"\" width=\"531\" height=\"347\" srcset=\"https:\/\/bionauts.jp\/wp-content\/uploads\/2025\/08\/WL-05177.png 531w, https:\/\/bionauts.jp\/wp-content\/uploads\/2025\/08\/WL-05177-300x196.png 300w\" sizes=\"(max-width: 531px) 85vw, 531px\" \/><\/a><\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n<h3>Background and Technology<\/h3>\n<p>To overcome the limitations of protein expression in <em>Streptomyces<\/em>, we successfully introduced a high-expression system derived from <em>Rhodococcus<\/em> into <em>Streptomyces<\/em> hosts. The expression system in <em>Rhodococcus<\/em> is industrially validated, exhibiting robust transcriptional activity sufficient to produce low-cost commodity chemicals such as acrylamide.<\/p>\n<p>Specifically, we introduced the transcriptional regulators NhhC and NhhD, along with the N-Hase promoter from the <em>Rhodococcus<\/em> nitrile hydratase gene cluster, into <em>Streptomyces lividans<\/em>. This demonstrated that the <em>Rhodococcus<\/em>-based system functions in <em>Streptomyces<\/em>, leading to the development of a high-expression vector, pHSA81.<\/p>\n<p>To further enable efficient extracellular secretion, we incorporated Tat secretion signal sequences from <em>S. avermitilis<\/em> and modified the vector to replicate in <em>E. coli<\/em>, resulting in the construction of a shuttle vector, pEHSA81k-ss1053, optimized for use in <em>Streptomyces<\/em>. Secretion efficiency was evaluated and confirmed. Additionally, we developed seven other expression vectors containing alternative Tat signal peptides (besides pEHSA81k-ss1053), enabling flexible adaptation depending on the target protein.<\/p>\n<h3>Data<\/h3>\n<ul>\n<li>Successful expression and secretion demonstrated for enzymes such as Pullulanase, Esterase, Chitinase, Phospholipase A2, and Amylase.<\/li>\n<li>In some cases, the target protein accounted for over two-thirds of the total secreted proteins in the culture supernatant.<\/li>\n<\/ul>\n<table style=\"border-collapse: collapse; width: 53.1667%; height: 413px;\">\n<tbody>\n<tr>\n<td style=\"width: 100%;\"><a href=\"http:\/\/bionauts.jp\/wp-content\/uploads\/2025\/08\/WL-05177b.png\"><img decoding=\"async\" loading=\"lazy\" class=\"alignleft size-full wp-image-4722\" src=\"http:\/\/bionauts.jp\/wp-content\/uploads\/2025\/08\/WL-05177b.png\" alt=\"\" width=\"531\" height=\"542\" srcset=\"https:\/\/bionauts.jp\/wp-content\/uploads\/2025\/08\/WL-05177b.png 531w, https:\/\/bionauts.jp\/wp-content\/uploads\/2025\/08\/WL-05177b-294x300.png 294w\" sizes=\"(max-width: 531px) 85vw, 531px\" \/><\/a><\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n<h3>Patent<\/h3>\n<p>JPB 006906215<\/p>\n<h3>Researcher<\/h3>\n<p>Dr. Michihiko Kobayashi (University of Tsukuba)<\/p>\n<h3>Expectations<\/h3>\n<p>We are seeking partners interested in adopting this expression vector system for protein production. Material Transfer Agreements (MTA) for technical evaluation are available.<\/p>\n<p>&nbsp;<\/p>\n<p>Project ID:WL-05177<\/p>\n<p>&nbsp;<\/p>\n","protected":false},"excerpt":{"rendered":"<p>High-Yield Protein Production Platform<\/p>\n","protected":false},"author":1,"featured_media":0,"comment_status":"closed","ping_status":"open","sticky":false,"template":"","format":"standard","meta":{"_mi_skip_tracking":false,"_monsterinsights_sitenote_active":false,"_monsterinsights_sitenote_note":"","_monsterinsights_sitenote_category":0},"categories":[1],"tags":[],"categories_of_inventions":[48],"disease_categories___indication":[],"asset_type":[],"development_phase":[],"medicine":[],"other_than_medicine":[131,130,133],"material":[],"engineering":[],"software_it":[],"aioseo_notices":[],"_links":{"self":[{"href":"https:\/\/bionauts.jp\/index.php?rest_route=\/wp\/v2\/posts\/4720"}],"collection":[{"href":"https:\/\/bionauts.jp\/index.php?rest_route=\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/bionauts.jp\/index.php?rest_route=\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/bionauts.jp\/index.php?rest_route=\/wp\/v2\/users\/1"}],"replies":[{"embeddable":true,"href":"https:\/\/bionauts.jp\/index.php?rest_route=%2Fwp%2Fv2%2Fcomments&post=4720"}],"version-history":[{"count":4,"href":"https:\/\/bionauts.jp\/index.php?rest_route=\/wp\/v2\/posts\/4720\/revisions"}],"predecessor-version":[{"id":4726,"href":"https:\/\/bionauts.jp\/index.php?rest_route=\/wp\/v2\/posts\/4720\/revisions\/4726"}],"wp:attachment":[{"href":"https:\/\/bionauts.jp\/index.php?rest_route=%2Fwp%2Fv2%2Fmedia&parent=4720"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/bionauts.jp\/index.php?rest_route=%2Fwp%2Fv2%2Fcategories&post=4720"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/bionauts.jp\/index.php?rest_route=%2Fwp%2Fv2%2Ftags&post=4720"},{"taxonomy":"categories_of_inventions","embeddable":true,"href":"https:\/\/bionauts.jp\/index.php?rest_route=%2Fwp%2Fv2%2Fcategories_of_inventions&post=4720"},{"taxonomy":"disease_categories___indication","embeddable":true,"href":"https:\/\/bionauts.jp\/index.php?rest_route=%2Fwp%2Fv2%2Fdisease_categories___indication&post=4720"},{"taxonomy":"asset_type","embeddable":true,"href":"https:\/\/bionauts.jp\/index.php?rest_route=%2Fwp%2Fv2%2Fasset_type&post=4720"},{"taxonomy":"development_phase","embeddable":true,"href":"https:\/\/bionauts.jp\/index.php?rest_route=%2Fwp%2Fv2%2Fdevelopment_phase&post=4720"},{"taxonomy":"medicine","embeddable":true,"href":"https:\/\/bionauts.jp\/index.php?rest_route=%2Fwp%2Fv2%2Fmedicine&post=4720"},{"taxonomy":"other_than_medicine","embeddable":true,"href":"https:\/\/bionauts.jp\/index.php?rest_route=%2Fwp%2Fv2%2Fother_than_medicine&post=4720"},{"taxonomy":"material","embeddable":true,"href":"https:\/\/bionauts.jp\/index.php?rest_route=%2Fwp%2Fv2%2Fmaterial&post=4720"},{"taxonomy":"engineering","embeddable":true,"href":"https:\/\/bionauts.jp\/index.php?rest_route=%2Fwp%2Fv2%2Fengineering&post=4720"},{"taxonomy":"software_it","embeddable":true,"href":"https:\/\/bionauts.jp\/index.php?rest_route=%2Fwp%2Fv2%2Fsoftware_it&post=4720"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}