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Journal Article 9

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site-directed mutagenesis 3

ARTP mutagenesis 2

1-naphthol 1

4-aminoantipyrine 1

Bacillus cereus QCG 1

Bacillus licheniformis 1

Sporisorium reilianum 1

?-poly-L-lysine 1

Streptomyces diastatochromogenes 1

AHV 1

Bioremediation 1

Enzyme immobilization 1

Lipase 1

Molecular dynamics simulation 1

Moxifloxacin 1

NADH-dependent enzyme 1

NADPH-dependent enzyme 1

Protein engineering 1

Rational design 1

Site-directed mutagenesis 1

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Combination of ARTP mutagenesis and color-mediated high-throughput screening to enhance 1-naphthol yield

Chenggang Qiu, Alei Zhang, Sha Tao, Kang Li, Kequan Chen, Pingkai Ouyang

Frontiers of Chemical Science and Engineering 2020, Volume 14, Issue 5,   Pages 793-801 doi: 10.1007/s11705-019-1876-2

Abstract: QCG was mutagenized to enhance 1-naphthol production, using atmospheric and room temperature plasma (ARTP

Keywords： Bacillus cereus QCG     naphthalene     1-naphthol     ARTP mutagenesis     high-throughput screening     4-aminoantipyrine    

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analysis of with enhanced poly--glutamic acid production through atmospheric and room temperature plasma mutagenesis

Frontiers of Chemical Science and Engineering 2022, Volume 16, Issue 12,   Pages 1751-1760 doi: 10.1007/s11705-022-2211-x

Abstract: Poly-γ-glutamic acid is an extracellular polymeric substance with various applications owing to its valuable properties of biodegradability, flocculating activity, water solubility, and nontoxicity. However, the ability of natural strains to produce poly-γ-glutamic acid is low. Atmospheric and room temperature plasma was applied in this study to conduct mutation breeding of Bacillus licheniformis CGMCC 2876, and a mutant strain M32 with an 11% increase in poly-γ-glutamic acid was obtained. Genome resequencing analysis identified 7 nonsynonymous mutations of ppsC encoding lipopeptide synthetase associated with poly-γ-glutamic acid metabolic pathways. From molecular docking, more binding sites and higher binding energy were speculated between the mutated plipastatin synthase subunit C and glutamate, which might contribute to the higher poly-γ-glutamic acid production. Moreover, the metabolic mechanism analysis revealed that the upregulated amino acids of M32 provided substrates for glutamate and promoted the conversion between L- and D-glutamate acids. In addition, the glycolytic pathway is enhanced, leading to a better capacity for using glucose. The maximum poly-γ-glutamic acid yield of 14.08 g·L–1 was finally reached with 30 g·L–1 glutamate.

Keywords： ARTP mutagenesis     Bacillus licheniformis     poly-γ-glutamic acid     metabolomics    

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Stem cell gene therapy: the risks of insertional mutagenesis and approaches to minimize genotoxicity

Chuanfeng Wu, Cynthia E. Dunbar

Frontiers of Medicine 2011, Volume 5, Issue 4,   Pages 356-371 doi: 10.1007/s11684-011-0159-1

Abstract: This review will summarize current information on the mechanisms of insertional mutagenesis in hematopoietic

Keywords： gene therapy     hematopoietic stem cells     insertional mutagenesis     genotoxicity     induced pluripotent stem cell    

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Site-directed mutagenesis of long QT syndrome KCNQ1 gene

LI Wei, WANG Bin, XU Qiumei, KE Qinmei, YANG Junguo, DU Rong, TIAN Li, WANG Qing

Frontiers of Medicine 2008, Volume 2, Issue 1,   Pages 100-104 doi: 10.1007/s11684-008-0018-x

Abstract: To construct a polymerase chain reaction (PCR) site-directed mutagenesis of the long QT syndrome KCNQ1Mutagenesis was performed in a two-step PCR.

Keywords： restriction     digestion     syndrome     sequence     site-directed mutagenesis    

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Mutagenesis and selective breeding of a high producing

Tian WANG, Shiru JIA, Zhilei TAN, Yujie DAI, Shuai SONG, Guoliang WANG

Frontiers of Chemical Science and Engineering 2012, Volume 6, Issue 2,   Pages 179-183 doi: 10.1007/s11705-012-1273-6

Abstract: -Poly- -lysine ( -PL) is an -lysine linear homopolymer, which is produced by bacteria belonging to the family and by ergot fungi. However, the production of -PL by the wild bacteria strain is very low, which limits its utilization. In most bacteria including the genus, -lysine is a precursor of -PL and is biosynthesized by the -aspartate pathway. Aspartokinase (Ask) is the first key enzyme in this pathway and is subject to complex regulation such as the feedback inhibition by the end product amino acids. In addition, phosphoenolpyruvate carboxykinase is feedback-regulated by -aspartate. To reduce these feedback inhibitions and to improve -PL productivity, resistant mutants were produced using sulfaguanidine (SG) + glycine+ -lysine+ -3-hydroxynorvaline (AHV) as selective markers. Using the interaction between -PL and the charged dye in the solid culture medium, hundreds of colonies were simultaneously screened in a quick and effective manner. Finally, one -PL-producing strain, L9, was selected. The productivity of this strain during flask fermentation was 0.77 g/L, which was 15% higher than that of the original strain. Moreover, its fermentation performance and genetic characteristics were very stable.

Keywords： ?-poly-L-lysine     plasma     AHV     Streptomyces diastatochromogenes     fermentation    

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Cofactor engineering in cyanobacteria to overcome imbalance between NADPH and NADH: A mini review

Jongmoon Park,Yunnam Choi

Frontiers of Chemical Science and Engineering 2017, Volume 11, Issue 1,   Pages 66-71 doi: 10.1007/s11705-016-1591-1

Abstract: Cyanobacteria can produce useful renewable fuels and high-value chemicals using sunlight and atmospheric carbon dioxide by photosynthesis. Genetic manipulation has increased the variety of chemicals that cyanobacteria can produce. However, their uniquely abundant NADPH-pool, in other words insufficient supply of NADH, tends to limit their production yields in case of utilizing NADH-dependent enzyme, which is quite common in heterotrophic microbes. To overcome this cofactor imbalance and enhance cyanobacterial fuel and chemical production, various approaches for cofactor engineering have been employed. In this review, we focus on three approaches: (1) utilization of NADPH-dependent enzymes, (2) increasing NADH production, and (3) changing cofactor specificity of NADH-dependent enzymes from NADH to NADPH.

Keywords： NADH-dependent enzyme     NADPH-dependent enzyme     transhydrogenase     site-directed mutagenesis     enzyme engineering    

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Improved degradation of azo dyes by lignin peroxidase following mutagenesis at two sites near the catalytic

Karla Ilić Đurđić, Raluca Ostafe, Olivera Prodanović, Aleksandra Đurđević Đelmaš, Nikolina Popović, Rainer Fischer, Stefan Schillberg, Radivoje Prodanović

Frontiers of Environmental Science & Engineering 2021, Volume 15, Issue 2, doi: 10.1007/s11783-020-1311-4

Abstract: subjected two positions (165 and 264) in the environment of the catalytic Trp171 residue to saturation mutagenesis

Keywords： Bioremediation     Enzyme immobilization     Protein engineering     Yeast surface display.    

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Rational Design, Over-expression and Characterization of a Novel Staphylokinase with Lower Tendency of Polymerization

Song Gang,Yu Min,Mo Wei,Song Houyan

Strategic Study of CAE 2000, Volume 2, Issue 11,   Pages 68-72

Abstract: RGD-Sak gene was constructed by substitution mutagenesis of Lys-109 and Phe-111 with Arg-109 and Asp

Keywords： staphylokinase     dimer     molecular docking     molecular design     site-directed mutagenesis    

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Efficient Activity Enhancement of a Lipase from Sporisorium reilianum for the Synthesis of a Moxifloxacin Chiral Intermediate via Rational Design Article

Xue Cai, Jiang-Wei Shen, Yu Qiang, Jing Hua, Zhang-Qi Ma, Zhi-Qiang Liu, Yu-Guo Zheng

Engineering 2022, Volume 19, Issue 12,   Pages 207-216 doi: 10.1016/j.eng.2022.03.020

Abstract:

Lipase-catalyzed stereoselective resolution of cis-(±)-dimethyl 1-acetylpiperidine-2,3-dicarboxylate (cis-(±)-1) is an attractive route for the synthesis of (S,S)-2,8-diazobicyclo[4.3.0]nonane, an important chiral intermediate of the fluoroquinolone antibiotic, moxifloxacin. In our previous study, a lipase from Sporisorium reilianum (SRL) was identified to possess excellent thermostability and pH stability. However, the low enzymatic activity of the SRL is a challenge that must be addressed. A rational design was initially employed for SRL tailoring according to the engineered Candida antarctica lipase B (CALB), resulting in a beneficial variant called SRL-I194N/V195L. Subsequently, two key amino acid residues in loop 6, L145 and L154, which might modulate the lid conformation between open and closed, were identified. A tetra-site variant, SRL-I194N/V195L/L145V/L154G (V13), with a significantly enhanced activity of 87.8 U∙mg−1 was obtained; this value was 2195-fold higher than that of wild-type SRL. Variant V13 was used to prepare optically pure (2S,3R)-dimethyl 1-acetylpiperidine-2,3-dicarboxylate ((2S,3R)-1), resolving 1 mol∙L−1 cis-(±)-1 with a conversion of 49.9% in 2 h and absolute stereoselectivity (E > 200). Excellent stability with a half-life of 92.5 h was also observed at 50 °C. Overall, the study findings reveal a lipase with high activity toward cis-(±)-1 at an industrial level and may offer a general strategy for enhancing the enzyme activity of other lipases and other classes of enzymes with a lid moiety.

Keywords： Lipase     Sporisorium reilianum     Site-directed mutagenesis     Molecular dynamics simulation     Rational    

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Title Author Date Type Operation

Combination of ARTP mutagenesis and color-mediated high-throughput screening to enhance 1-naphthol yield

Chenggang Qiu, Alei Zhang, Sha Tao, Kang Li, Kequan Chen, Pingkai Ouyang

Journal Article

analysis of with enhanced poly--glutamic acid production through atmospheric and room temperature plasma mutagenesis

Journal Article

Stem cell gene therapy: the risks of insertional mutagenesis and approaches to minimize genotoxicity

Chuanfeng Wu, Cynthia E. Dunbar

Journal Article

Site-directed mutagenesis of long QT syndrome KCNQ1 gene

LI Wei, WANG Bin, XU Qiumei, KE Qinmei, YANG Junguo, DU Rong, TIAN Li, WANG Qing

Journal Article

Mutagenesis and selective breeding of a high producing

Tian WANG, Shiru JIA, Zhilei TAN, Yujie DAI, Shuai SONG, Guoliang WANG

Journal Article

Cofactor engineering in cyanobacteria to overcome imbalance between NADPH and NADH: A mini review

Jongmoon Park,Yunnam Choi

Journal Article

Improved degradation of azo dyes by lignin peroxidase following mutagenesis at two sites near the catalytic

Karla Ilić Đurđić, Raluca Ostafe, Olivera Prodanović, Aleksandra Đurđević Đelmaš, Nikolina Popović, Rainer Fischer, Stefan Schillberg, Radivoje Prodanović

Journal Article

Rational Design, Over-expression and Characterization of a Novel Staphylokinase with Lower Tendency of Polymerization

Song Gang,Yu Min,Mo Wei,Song Houyan

Journal Article

Efficient Activity Enhancement of a Lipase from Sporisorium reilianum for the Synthesis of a Moxifloxacin Chiral Intermediate via Rational Design

Xue Cai, Jiang-Wei Shen, Yu Qiang, Jing Hua, Zhang-Qi Ma, Zhi-Qiang Liu, Yu-Guo Zheng

Journal Article