促进甘油对大肠杆菌重组蛋白产生影响的诱导条件

Q1 Immunology and Microbiology

Biotechnology Reports Pub Date : 2025-05-16 DOI:10.1016/j.btre.2025.e00898

Yoshihiro Ojima , Hajime Saito , Shintaro Miyuki , Koichi Fukunaga , Terumichi Tsuboi , Masayuki Azuma

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引用次数: 0

摘要

用pET系统在含有葡萄糖（Glu培养基）、葡萄糖和甘油（Glu培养基）或甘油（Gly培养基）的最小培养基中表达胰岛素原菌。当IPTG为100 μM时，胰岛素原的生成不随甘油的加入而增加。相比之下，在GluGly和Gly培养基中，每介质体积的胰岛素原产量比在10 μM IPTG的Glu培养基中大约高3 ~ 4倍。与Glu培养基相比，Glu培养基中靶蛋白的mRNA表达更高，表明释放葡萄糖诱导的分解代谢抑制物增强了胰岛素原的产生。虽然GluGly和Gly培养基在25 h时的胰岛素原产量没有差异，但在GluGly培养基中底物消耗很快，15h时胰岛素原产量高出1.55±0.12倍。考虑到生产周期，GluGly培养基的产量最高。本研究表明葡萄糖和甘油的混合物在低IPTG浓度的大肠杆菌中生产蛋白质是有价值的。

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Induction conditions that promote the effect of glycerol on recombinant protein production in Escherichia coli

Proinsulin was expressed by Escherichia coli SHuffle T7 with pET system in minimal medium containing glucose (Glu medium), glucose and glycerol (GluGly medium) or glycerol (Gly medium). With 100 μM IPTG, proinsulin production did not increase with glycerol. In contrast, proinsulin production per medium volume in GluGly and Gly media was approximately 3∼4-fold higher than in Glu medium with 10 μM IPTG. mRNA expression of target protein was higher in GluGly versus Glu medium, indicating that proinsulin production was enhanced by the release of glucose-induced catabolite inhibition. Although proinsulin production did not differ between GluGly and Gly media at 25 h, substrate was consumed quickly in GluGly medium with 1.55±0.12 times higher proinsulin production at 15 h. Productivity, considering the production period, was highest in the GluGly medium. This study shows a mixture of glucose and glycerol is valuable for protein production in E. coli with low IPTG concentrations.

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来源期刊

Biotechnology Reports Immunology and Microbiology-Applied Microbiology and Biotechnology

CiteScore

15.80

自引率

0.00%

发文量

审稿时长

55 days

期刊介绍： Biotechnology Reports covers all aspects of Biotechnology particularly those reports that are useful and informative and that will be of value to other researchers in related fields. Biotechnology Reports loves ground breaking science, but will also accept good science that can be of use to the biotechnology community. The journal maintains a high quality peer review where submissions are considered on the basis of scientific validity and technical quality. Acceptable paper types are research articles (short or full communications), methods, mini-reviews, and commentaries in the following areas: Healthcare and pharmaceutical biotechnology Agricultural and food biotechnology Environmental biotechnology Molecular biology, cell and tissue engineering and synthetic biology Industrial biotechnology, biofuels and bioenergy Nanobiotechnology Bioinformatics & systems biology New processes and products in biotechnology, bioprocess engineering.