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      Indirect Pathway Metabolic Engineering Strategies for Enhanced Biosynthesis of Hyaluronic Acid in Engineered Corynebacterium glutamicum

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          Abstract

          Hyaluronic acid (HA) is composed of alternating d-glucuronic acid and N-acetyl- d-glucosamine, with excellent biocompatibility and water retention capacity. To achieve heterologous biosynthesis of HA, Corynebacterium glutamicum, a safe GRAS (generally recognized as safe) host, was utilized and metabolically engineered previously. In this work, to achieve further enhancement of HA yield, four strategies were proposed and performed separately first, i.e., (1) improvement of glucose uptake via iolR gene knockout, releasing the inhibition of transporter IolT1/IolT2 and glucokinases; (2) intensification of cardiolipin synthesis through overexpression of genes pgsA1/ pgsA2/ cls involved in cardiolipin synthesis; (3) duly expressed Vitreoscilla hemoglobin in genome, enhancing HA titer coupled with more ATP and improved NAD +/NADH (>7.5) ratio; and (4) identification of the importance of glutamine for HA synthesis through transcriptome analyses and then enhancement of the HA titer via its supplement. After that, we combined different strategies together to further increase the HA titer. As a result, one of the optimal recombinant strains, Cg-dR-CLS, yielded 32 g/L of HA at 60 h in a fed-batch culture, which was increased by 30% compared with that of the starting strain. This high value of HA titer will enable the industrial production of HA via the engineered C. glutamicum.

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          Most cited references60

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          Small mobilizable multi-purpose cloning vectors derived from the Escherichia coli plasmids pK18 and pK19: selection of defined deletions in the chromosome of Corynebacterium glutamicum

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            Hyaluronic acid—Based wound dressings: A review

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              Molecular basis for membrane phospholipid diversity: why are there so many lipids?

              W Dowhan (1996)
              Phospholipids play multiple roles in cells by establishing the permeability barrier for cells and cell organelles, by providing the matrix for the assembly and function of a wide variety of catalytic processes, by acting as donors in the synthesis of macromolecules, and by actively influencing the functional properties of membrane-associated processes. The function, at the molecular level, of phosphatidylethanolamine, phosphatidylglycerol, and cardiolipin in specific cellular processes is reviewed, with a focus on the results of combined molecular genetic and biochemical studies in Escherichia coli. These results are compared with primarily biochemical data supporting similar functions for these phospholipids in eukaryotic organisms. The wide range of processes in which specific involvement of phospholipids has been documented explains the need for diversity in phospholipid structure and why there are so many membrane lipids.
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                Author and article information

                Contributors
                Journal
                Front Bioeng Biotechnol
                Front Bioeng Biotechnol
                Front. Bioeng. Biotechnol.
                Frontiers in Bioengineering and Biotechnology
                Frontiers Media S.A.
                2296-4185
                20 December 2021
                2021
                : 9
                : 768490
                Affiliations
                [1] 1 Key Laboratory for Industrial Biocatalysis of the Ministry of Education, Department of Chemical Engineering, Tsinghua University , Beijing, China
                [2] 2 Center for Synthetic and Systems Biology, Tsinghua University , Beijing, China
                Author notes

                Edited by: Yu Wang, Tianjin Institute of Industrial Biotechnology (CAS), China

                Reviewed by: Yu-Sin Jang, Gyeongsang National University, South Korea

                Yota Tsuge, Kanazawa University, Japan

                *Correspondence: Huimin Yu, yuhm@ 123456tsinghua.edu.cn

                This article was submitted to Synthetic Biology, a section of the journal Frontiers in Bioengineering and Biotechnology

                Article
                768490
                10.3389/fbioe.2021.768490
                8721151
                ac887cca-0cf1-4587-b150-172be57c38d3
                Copyright © 2021 Du, Cheng, Wang, Xu and Yu.

                This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

                History
                : 31 August 2021
                : 22 November 2021
                Categories
                Bioengineering and Biotechnology
                Original Research

                hyaluronic acid,engineered corynebacterium glutamicum ,iolr deletion,cardiolipin (cl), vitreoscilla hemoglobin (vhb),glutamine

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