Enhancing human‐like collagen accumulation by deleting the major glucose transporter ptsG in recombinant Escherichia coli BL21. (11th March 2014)

• Record Type: Journal Article
• Authors: Luo, Yan'e; Zhang, Tao; Fan, Daidi; Mu, Tingzhen; Xue, Wenjiao; Hui, Junfeng; Ma, Xiaoxuan
• Is Part Of: Biotechnology and applied biochemistry. Volume 61:Number 2(2014)
• Page Start: 237
• Page End: 247
• Publication Date: 2014-03-11
• Subjects: Biotechnology -- Periodicals; Biochemical engineering -- Periodicals; Biochemistry -- Periodicals; Biochemistry -- Periodicals; Genetic Techniques -- Periodicals; Microbiological Techniques -- Periodicals; 660.6
• DOI: 10.1002/bab.1171
• Languages: English
• ISSNs: 0885-4513

Abstract

<abstract abstract-type="main"> <title>Abstract</title> <p>Collagen has been proven to be a valuable biomedical material for many medical applications. Human‐like collagen (HLC) is a novel important biomedical material with diverse medical applications. In this work, recombinant <italic>Escherichia coli</italic> BL21 3.7 ∆<italic>ptsG</italic> was constructed, the characters of <italic>ptsG</italic> mutant strain were analyzed, and real‐time quantitative polymerase chain reaction (PCR) was applied to investigate the effect of <italic>ptsG</italic> gene deletion on the transcriptional level of the phosphotransferase system (PTS) genes responsible for glucose transport. The HLC production and cell growth ability were 1.33‐ and 1.24‐fold higher than those of its parent strain in the fermentation medium, respectively, and 1.16‐ and 1.17‐fold in the modified minimal medium individually. The acetate accumulation decreased by 42%–56% compared to its parent strain in the fermentation medium, and 70%–87% in the modified minimal medium. The results of RT‐qPCR showed that the transcriptional level of <italic>crr</italic>, <italic>ptsH</italic>, <italic>ptsI</italic>, and <italic>blgF</italic> in <italic>ptsG</italic> mutant all decreased dramatically, which inferred a decrease in the glucose uptake rate, but the transcriptional level of <italic>FruB</italic> and <italic>manX</italic> increased slightly, which demonstrated the activation of fructose‐ and mannose‐specific transport pathways in the <italic>ptsG</italic> mutant. This study demonstrates that <italic>ptsG</italic> deletion is an effective strategy to reduce acetate accumulation and increase biomass and HLC production.</p> </abstract>