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Wendisch, Volker F. - One of the best experts on this subject based on the ideXlab platform.
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Dynamic co-cultivation process of Corynebacterium glutamicum strains for the fermentative production of riboflavin
'MDPI AG', 2021Co-Authors: Perez Fernando, Wendisch, Volker F., Burgardt Arthur, Kallmann, Dina R., Bar NadavAbstract:Perez F, Burgardt A, Kallmann DR, Wendisch VF, Bar N. Dynamic co-cultivation process of Corynebacterium glutamicum strains for the fermentative production of riboflavin. Fermentation. 2021;7(1): 11
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Adaptive Laboratory Evolution accelerated glutarate production by Corynebacterium glutamicum
'Springer Science and Business Media LLC', 2021Co-Authors: Prell Carina, Busche Tobias, Rückert Christian, Nolte Lea, Brandenbusch Christoph, Wendisch, Volker F.Abstract:Prell C, Busche T, Rückert C, Nolte L, Brandenbusch C, Wendisch VF. Adaptive Laboratory Evolution accelerated glutarate production by Corynebacterium glutamicum. Microbial Cell Factories. 2021;20:97
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Improved plasmid-based inducible and constitutive gene expression in Corynebacterium glutamicum
'MDPI AG', 2021Co-Authors: Henke, Nadja Alina, Krahn Irene, Wendisch, Volker F.Abstract:Henke NA, Krahn I, Wendisch VF. Improved plasmid-based inducible and constitutive gene expression in Corynebacterium glutamicum. Microorganisms. 2021;9(1): 204.Corynebacterium glutamicum has been safely used in white biotechnology for the last 60 years and the portfolio of new pathways and products is increasing rapidly. Hence, expression vectors play a central role in discovering endogenous gene functions and in establishing heterologous gene expression. In this work, new expression vectors were designed based on two strategies: (i) a library screening of constitutive native and synthetic promoters and (ii) an increase of the plasmid copy number. Both strategies were combined and resulted in a very strong expression and overproduction of the fluorescence protein GfpUV. As a second test case, the improved vector for constitutive expression was used to overexpress the endogenous xylulokinase gene xylB in a synthetic operon with xylose isomerase gene xylA from Xanthomonas campestris. The xylose isomerase activity in crude extracts was increased by about three-fold as compared to that of the parental vector. In terms of application, the improved vector for constitutive xylA and xylB expression was used for production of the N-methylated amino acid sarcosine from monomethylamine, acetate, and xylose. As a consequence, the volumetric productivity of sarcosine production was 50% higher as compared to that of the strain carrying the parental vector
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Fermentative production of ʟ-2-hydroxyglutarate by engineered Corynebacterium glutamicum via pathway extension of ʟ-lysine biosynthesis
'Frontiers Media SA', 2021Co-Authors: Prell Carina, Burgardt Arthur, Meyer Florian, Wendisch, Volker F.Abstract:Prell C, Burgardt A, Meyer F, Wendisch VF. Fermentative production of ʟ-2-hydroxyglutarate by engineered Corynebacterium glutamicum via pathway extension of ʟ-lysine biosynthesis. Frontiers in Bioengineering and Biotechnology. Accepted
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Evolving a new efficient mode of fructose utilization for improved bioproduction in Corynebacterium glutamicum
'Frontiers Media SA', 2021Co-Authors: Krahn Irene, Wendisch, Volker F., Bonder Daniel, Torregrosa Lucia, Stoppel Dominik, Krause Jens, Rosenfeldt Natalie, Meiswinkel Tobias, Seibold, Gerd M., Lindner SteffenAbstract:Krahn I, Bonder D, Torregrosa L, et al. Evolving a new efficient mode of fructose utilization for improved bioproduction in Corynebacterium glutamicum. Frontiers in Biotechnology and Bioengineering. 2021;9: 669093
Sheng Yang - One of the best experts on this subject based on the ideXlab platform.
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crispr cpf1 assisted genome editing of Corynebacterium glutamicum
Nature Communications, 2017Co-Authors: Yu Jiang, Fenghui Qian, Junjie Yang, Yingmiao Liu, Feng Dong, Bingbing Sun, Biao Chen, Renxiao Wang, Sheng YangAbstract:Corynebacterium glutamicum is an important industrial metabolite producer that is difficult to genetically engineer. Although the Streptococcus pyogenes (Sp) CRISPR-Cas9 system has been adapted for genome editing of multiple bacteria, it cannot be introduced into C. glutamicum. Here we report a Francisella novicida (Fn) CRISPR-Cpf1-based genome-editing method for C. glutamicum. CRISPR-Cpf1, combined with single-stranded DNA (ssDNA) recombineering, precisely introduces small changes into the bacterial genome at efficiencies of 86-100%. Large gene deletions and insertions are also obtained using an all-in-one plasmid consisting of FnCpf1, CRISPR RNA, and homologous arms. The two CRISPR-Cpf1-assisted systems enable N iterative rounds of genome editing in 3N+4 or 3N+2 days. A proof-of-concept, codon saturation mutagenesis at G149 of γ-glutamyl kinase relieves L-proline inhibition using Cpf1-assisted ssDNA recombineering. Thus, CRISPR-Cpf1-based genome editing provides a highly efficient tool for genetic engineering of Corynebacterium and other bacteria that cannot utilize the Sp CRISPR-Cas9 system.
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crispr cpf1 assisted genome editing of Corynebacterium glutamicum
Nature Communications, 2017Co-Authors: Yu Jiang, Fenghui Qian, Junjie Yang, Feng Dong, Biao Chen, Renxiao Wang, Chongmao Xu, Xiaoshu Xu, Yan Li, Sheng YangAbstract:Corynebacterium glutamicum is an important industrial metabolite producer that is difficult to genetically engineer. Although the Streptococcus pyogenes (Sp) CRISPR-Cas9 system has been adapted for genome editing of multiple bacteria, it cannot be introduced into C. glutamicum. Here we report a Francisella novicida (Fn) CRISPR-Cpf1-based genome-editing method for C. glutamicum. CRISPR-Cpf1, combined with single-stranded DNA (ssDNA) recombineering, precisely introduces small changes into the bacterial genome at efficiencies of 86–100%. Large gene deletions and insertions are also obtained using an all-in-one plasmid consisting of FnCpf1, CRISPR RNA, and homologous arms. The two CRISPR-Cpf1-assisted systems enable N iterative rounds of genome editing in 3N+4 or 3N+2 days. A proof-of-concept, codon saturation mutagenesis at G149 of γ-glutamyl kinase relieves L-proline inhibition using Cpf1-assisted ssDNA recombineering. Thus, CRISPR-Cpf1-based genome editing provides a highly efficient tool for genetic engineering of Corynebacterium and other bacteria that cannot utilize the Sp CRISPR-Cas9 system. Corynebacterium glutamicum is an important industrial microbe, however it has proven difficult to genetically engineer using Cas9 from Streptococcus pyogenes. Here the authors report effective genome engineering of the bacterium using Cpf1 from Francisella novicida.
Hideaki Yukawa - One of the best experts on this subject based on the ideXlab platform.
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Characterization of shikimate dehydrogenase homologues of Corynebacterium glutamicum
Applied Microbiology and Biotechnology, 2013Co-Authors: Takeshi Kubota, Yuya Tanaka, Kazumi Hiraga, Masayuki Inui, Hideaki YukawaAbstract:The function of three Corynebacterium glutamicum shikimate dehydrogenase homologues, designated as qsuD (cgR_0495), cgR_1216, and aroE (cgR_1677), was investigated. A disruptant of aroE required shikimate for growth, whereas a qsuD-deficient strain did not grow in medium supplemented with either quinate or shikimate as sole carbon sources. There was no discernible difference in growth rate between wild-type and a cgR_1216-deficient strain. Enzymatic assays showed that AroE both reduced 3-dehydroshikimate, using NADPH as cofactor, and oxidized shikimate, the reverse reaction, using NADP+ as cofactor. The reduction reaction was ten times faster than the oxidation. QsuD reduced 3-dehydroquinate using NADH and oxidized quinate using NAD+ as cofactor. Different from the other two homologues, the product of cgR_1216 displayed considerably lower enzyme activity for both the reduction and the oxidation. The catalytic reaction of QsuD and AroE was highly susceptible to pH. Furthermore, reduction of 3-dehydroshikimate by AroE was inhibited by high concentrations of shikimate, but neither quinate nor aromatic amino acids had any effect on the reaction. Expression of qsuD mRNA was strongly enhanced in the presence of shikimate, whereas that of cgR_1216 and aroE decreased. We conclude that while AroE is the main catalyst for shikimate production in the shikimate pathway, QsuD is essential for quinate/shikimate utilization.
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Xylitol production by recombinant Corynebacterium glutamicum under oxygen deprivation
Applied Microbiology and Biotechnology, 2010Co-Authors: Miho Sasaki, Masayuki Inui, Toru Jojima, Hideaki YukawaAbstract:Wild-type Corynebacterium glutamicum produced 0.6 g l^−1 xylitol from xylose at a productivity of 0.01 g l^−1 h^−1 under oxygen deprivation. To increase this productivity, the pentose transporter gene ( araE ) from C. glutamicum ATCC31831 was integrated into the C . glutamicum R chromosome. Consequent disruption of its lactate dehydrogenase gene ( ldhA ), and expression of single-site mutant xylose reductase from Candida tenuis ( CtXR (K274R)) resulted in recombinant C. glutamicum strain CtXR4 that produced 26.5 g l^−1 xylitol at 3.1 g l^−1 h^−1. To eliminate possible formation of toxic intracellular xylitol phosphate, genes encoding xylulokinase (XylB) and phosphoenolpyruvate-dependent fructose phosphotransferase (PTS^fru) were disrupted to yield strain CtXR7. The productivity of strain CtXR7 increased 1.6-fold over that of strain CtXR4. A fed-batch 21-h CtXR7 culture in mineral salts medium under oxygen deprivation yielded 166 g l^−1 xylitol at 7.9 g l^−1 h^−1, representing the highest bacterial xylitol productivity reported to date.
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transcription of Corynebacterium glutamicum genes involved in tricarboxylic acid cycle and glyoxylate cycle
Journal of Molecular Microbiology and Biotechnology, 2008Co-Authors: Sung Ok Han, Masayuki Inui, Hideaki YukawaAbstract:Transcription of the tricarboxylic acid cycle genes of Corynebacterium glutamicum was investigated. Northern hybridizations revealed that gltA-fkb, odhA-orfA, sucC-sucD, sd
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production of d lactic acid by Corynebacterium glutamicum under oxygen deprivation
Applied Microbiology and Biotechnology, 2005Co-Authors: Shohei Okino, Masayuki Inui, Masako Suda, Keitaro Fujikura, Hideaki YukawaAbstract:In mineral salts medium under oxygen deprivation, Corynebacterium glutamicum exhibits high productivity of l-lactic acid accompanied with succinic and acetic acids. In taking advantage of this elevated productivity, C. glutamicum was genetically modified to produce d-lactic acid. The modification involved expression of fermentative d-lactate dehydrogenase (d-LDH)-encoding genes from Escherichia coli and Lactobacillus delbrueckii in l-lactate dehydrogenase (l-LDH)-encoding ldhA-null C. glutamicum mutants to yield strains C. glutamicum ΔldhA/pCRB201 and C. glutamicum ΔldhA/pCRB204, respectively. The productivity of C. glutamicum ΔldhA/pCRB204 was fivefold higher than that of C. glutamicum ΔldhA/pCRB201. By using C. glutamicum ΔldhA/pCRB204 cells packed to a high density in mineral salts medium, up to 1,336 mM (120 g l−1) of d-lactic acid of greater than 99.9% optical purity was produced within 30 h.
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metabolic analysis of Corynebacterium glutamicum during lactate and succinate productions under oxygen deprivation conditions
Journal of Molecular Microbiology and Biotechnology, 2004Co-Authors: Masayuki Inui, Hideo Kawaguchi, Shikiko Murakami, Shohei Okino, Alain A Vertes, Hideaki YukawaAbstract:Lactate and succinate were produced from glucose by Corynebacterium glutamicum under oxygen deprivation conditions without growth. Addition of bicarbonate to the reaction mixture le
Yu Jiang - One of the best experts on this subject based on the ideXlab platform.
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crispr cpf1 assisted genome editing of Corynebacterium glutamicum
Nature Communications, 2017Co-Authors: Yu Jiang, Fenghui Qian, Junjie Yang, Yingmiao Liu, Feng Dong, Bingbing Sun, Biao Chen, Renxiao Wang, Sheng YangAbstract:Corynebacterium glutamicum is an important industrial metabolite producer that is difficult to genetically engineer. Although the Streptococcus pyogenes (Sp) CRISPR-Cas9 system has been adapted for genome editing of multiple bacteria, it cannot be introduced into C. glutamicum. Here we report a Francisella novicida (Fn) CRISPR-Cpf1-based genome-editing method for C. glutamicum. CRISPR-Cpf1, combined with single-stranded DNA (ssDNA) recombineering, precisely introduces small changes into the bacterial genome at efficiencies of 86-100%. Large gene deletions and insertions are also obtained using an all-in-one plasmid consisting of FnCpf1, CRISPR RNA, and homologous arms. The two CRISPR-Cpf1-assisted systems enable N iterative rounds of genome editing in 3N+4 or 3N+2 days. A proof-of-concept, codon saturation mutagenesis at G149 of γ-glutamyl kinase relieves L-proline inhibition using Cpf1-assisted ssDNA recombineering. Thus, CRISPR-Cpf1-based genome editing provides a highly efficient tool for genetic engineering of Corynebacterium and other bacteria that cannot utilize the Sp CRISPR-Cas9 system.
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crispr cpf1 assisted genome editing of Corynebacterium glutamicum
Nature Communications, 2017Co-Authors: Yu Jiang, Fenghui Qian, Junjie Yang, Feng Dong, Biao Chen, Renxiao Wang, Chongmao Xu, Xiaoshu Xu, Yan Li, Sheng YangAbstract:Corynebacterium glutamicum is an important industrial metabolite producer that is difficult to genetically engineer. Although the Streptococcus pyogenes (Sp) CRISPR-Cas9 system has been adapted for genome editing of multiple bacteria, it cannot be introduced into C. glutamicum. Here we report a Francisella novicida (Fn) CRISPR-Cpf1-based genome-editing method for C. glutamicum. CRISPR-Cpf1, combined with single-stranded DNA (ssDNA) recombineering, precisely introduces small changes into the bacterial genome at efficiencies of 86–100%. Large gene deletions and insertions are also obtained using an all-in-one plasmid consisting of FnCpf1, CRISPR RNA, and homologous arms. The two CRISPR-Cpf1-assisted systems enable N iterative rounds of genome editing in 3N+4 or 3N+2 days. A proof-of-concept, codon saturation mutagenesis at G149 of γ-glutamyl kinase relieves L-proline inhibition using Cpf1-assisted ssDNA recombineering. Thus, CRISPR-Cpf1-based genome editing provides a highly efficient tool for genetic engineering of Corynebacterium and other bacteria that cannot utilize the Sp CRISPR-Cas9 system. Corynebacterium glutamicum is an important industrial microbe, however it has proven difficult to genetically engineer using Cas9 from Streptococcus pyogenes. Here the authors report effective genome engineering of the bacterium using Cpf1 from Francisella novicida.
Renxiao Wang - One of the best experts on this subject based on the ideXlab platform.
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crispr cpf1 assisted genome editing of Corynebacterium glutamicum
Nature Communications, 2017Co-Authors: Yu Jiang, Fenghui Qian, Junjie Yang, Yingmiao Liu, Feng Dong, Bingbing Sun, Biao Chen, Renxiao Wang, Sheng YangAbstract:Corynebacterium glutamicum is an important industrial metabolite producer that is difficult to genetically engineer. Although the Streptococcus pyogenes (Sp) CRISPR-Cas9 system has been adapted for genome editing of multiple bacteria, it cannot be introduced into C. glutamicum. Here we report a Francisella novicida (Fn) CRISPR-Cpf1-based genome-editing method for C. glutamicum. CRISPR-Cpf1, combined with single-stranded DNA (ssDNA) recombineering, precisely introduces small changes into the bacterial genome at efficiencies of 86-100%. Large gene deletions and insertions are also obtained using an all-in-one plasmid consisting of FnCpf1, CRISPR RNA, and homologous arms. The two CRISPR-Cpf1-assisted systems enable N iterative rounds of genome editing in 3N+4 or 3N+2 days. A proof-of-concept, codon saturation mutagenesis at G149 of γ-glutamyl kinase relieves L-proline inhibition using Cpf1-assisted ssDNA recombineering. Thus, CRISPR-Cpf1-based genome editing provides a highly efficient tool for genetic engineering of Corynebacterium and other bacteria that cannot utilize the Sp CRISPR-Cas9 system.
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crispr cpf1 assisted genome editing of Corynebacterium glutamicum
Nature Communications, 2017Co-Authors: Yu Jiang, Fenghui Qian, Junjie Yang, Feng Dong, Biao Chen, Renxiao Wang, Chongmao Xu, Xiaoshu Xu, Yan Li, Sheng YangAbstract:Corynebacterium glutamicum is an important industrial metabolite producer that is difficult to genetically engineer. Although the Streptococcus pyogenes (Sp) CRISPR-Cas9 system has been adapted for genome editing of multiple bacteria, it cannot be introduced into C. glutamicum. Here we report a Francisella novicida (Fn) CRISPR-Cpf1-based genome-editing method for C. glutamicum. CRISPR-Cpf1, combined with single-stranded DNA (ssDNA) recombineering, precisely introduces small changes into the bacterial genome at efficiencies of 86–100%. Large gene deletions and insertions are also obtained using an all-in-one plasmid consisting of FnCpf1, CRISPR RNA, and homologous arms. The two CRISPR-Cpf1-assisted systems enable N iterative rounds of genome editing in 3N+4 or 3N+2 days. A proof-of-concept, codon saturation mutagenesis at G149 of γ-glutamyl kinase relieves L-proline inhibition using Cpf1-assisted ssDNA recombineering. Thus, CRISPR-Cpf1-based genome editing provides a highly efficient tool for genetic engineering of Corynebacterium and other bacteria that cannot utilize the Sp CRISPR-Cas9 system. Corynebacterium glutamicum is an important industrial microbe, however it has proven difficult to genetically engineer using Cas9 from Streptococcus pyogenes. Here the authors report effective genome engineering of the bacterium using Cpf1 from Francisella novicida.
