Site-directed mutagenesis of coenzyme-independent carotenoid oxygenase CSO2 to enhance the enzymatic synthesis of vanillin

Yao, Xueyan; Lv, Yuemeng; Yu, Hui‐Lei; Cao, Hao; Wang, Luyao; Wen, Bin; Gu, Tianyi; Wang, Fengzhong; Sun, Lichao; Xin, Fengjiao

doi:10.1007/s00253-020-10433-1

Cited by 20 publications

(15 citation statements)

References 28 publications

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“…With respect to protein solubility, the technology was utilized by biologists to identify nonfunctional hydrophobic residues exposed to the solvent where aggregation problems arose, and replace them by hydrophilic ones to increase surface polar interactions for the target protein. Pioneered in Alan Fersht’s lab in the 1980s, it has gradually become the most used technique to enhance protein solubility and stability. − …”

Section: Solubility and Introduction To Protein Designmentioning

confidence: 99%

Protein Design: From the Aspect of Water Solubility and Stability

et al. 2022

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Water solubility and structural stability are key merits for proteins defined by the primary sequence and 3D-conformation. Their manipulation represents important aspects of the protein design field that relies on the accurate placement of amino acids and molecular interactions, guided by underlying physiochemical principles. Emulated designer proteins with well-defined properties both fuel the knowledge-base for more precise computational design models and are used in various biomedical and nanotechnological applications. The continuous developments in protein science, increasing computing power, new algorithms, and characterization techniques provide sophisticated toolkits for solubility design beyond guess work. In this review, we summarize recent advances in the protein design field with respect to water solubility and structural stability. After introducing fundamental design rules, we discuss the transmembrane protein solubilization and de novo transmembrane protein design. Traditional strategies to enhance protein solubility and structural stability are introduced. The designs of stable protein complexes and high-order assemblies are covered. Computational methodologies behind these endeavors, including structure prediction programs, machine learning algorithms, and specialty software dedicated to the evaluation of protein solubility and aggregation, are discussed. The findings and opportunities for Cryo-EM are presented. This review provides an overview of significant progress and prospects in accurate protein design for solubility and stability.

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Section: Solubility and Introduction To Protein Designmentioning

confidence: 99%

Protein Design: From the Aspect of Water Solubility and Stability

et al. 2022

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“…In particular, it was previously reported that the low catalytic efficiency of Cso2 and its thermal instability could affect the vanillin production. 27 In our study, the amino acidic substitutions A49P and Q390A were introduced in Cso2 (Table 1), as suggested by the site-directed mutagenesis study performed by Yao et al, 28 and the high-copy number plasmid was used for its recombinant production.…”

Section: ■ Results and Discussionmentioning

confidence: 99%

Whole-Cell Bioconversion of Renewable Biomasses-Related Aromatics to cis,cis-Muconic Acid

Molinari

Pollegioni

Rosini

2023

ACS Sustainable Chem. Eng.

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Lignin and wheat bran represent renewable feedstocks for generation of useful and value-added compounds such as vanillin (a popular flavoring agent) and cis,cis-muconic acid (ccMA, a building block for the synthesis of plastic materials). In the present work, we report on the setup of an efficient and green process for producing such valuable compounds based on (a) the optimization of the extraction procedures for vanillin from lignin and ferulic acid from wheat bran and (b) the genetic engineering of an Escherichia coli strain with up to three plasmids differing in copy numbers to modulate the expression of up to seven recombinant enzymes. In detail, we used two sequential reactions catalyzed by the decarboxylase Fdc and the dioxygenase Ado to convert wheat bran-derived ferulic acid into vanillin: nature-identical vanillin was produced in one pot with a >85% yield in 20 h. Next, the dehydrogenase LigV, the demethylase VanAB, the decarboxylase AroY, and the dioxygenase C12O converted lignin-derived vanillin into ccMA with a >95% conversion yield and a productivity of 4.2 mg of ccMA/g of Kraft lignin in 30 min. Finally, when the optimized E. coli strain expressing all the abovementioned enzymes was used, ccMA was produced with a >95% conversion yield starting from ferulic acid in 10 h following product isolation, corresponding to 0.73 g of ccMA/g of ferulic acid, 1.4 g of ccMA/L, and 2.2 g of ccMA/g of wheat bran biomass. The optimized whole-cell system represents a sustainable and cost-competitive process for producing high value-added products from renewable resources.

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“…The content of FA in the samples was determined by high-performance liquid chromatography (HPLC) with a minor modification . The ethanol extract of the sample was prepared using the method described above.…”

Section: Methodsmentioning

confidence: 99%

“…The content of FA in the samples was determined by high-performance liquid chromatography (HPLC) with a minor modification. 25 The ethanol extract of the sample was prepared using the method described above. The extract was analyzed by a Shimadzu HPLC (model LC-10ATvp Pumps and DGU-12A Degasser, Shimadzu, Kyoto, Japan) equipped with a diode array detector (model SPD-M10Avp, Shimadzu, Kyoto, Japan).…”

Section: ■ Materials and Methodsmentioning

confidence: 99%

Process Optimization for Production of Ferulic Acid and Pentosans from Wheat Brans by Solid-State Fermentation and Evaluation of Their Antioxidant Activities

Zhang

Zhou

Zhang

et al. 2022

ACS Food Sci. Technol.

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Wheat bran (WB) is largely discarded during processing, resulting in wastage of the bioactive components in it, such as ferulic acid (FA) and pentosans. The aim of this study was to enhance their yield by solid-state fermentation with three filamentous fungi (Aspergillus oryzae, Rhizopus oryzae, and Aspergillus niger). Through single-factor experiments, the optimal fermentation conditions were found to be a strain of A. oryzae, an inoculation amount of A. oryzae of 4 × 10 5 spores/g, a moisture content of 60%, a fermentation temperature of 28 °C, and a fermentation time of 48 h. By 2,2-diphenyl-1-picrylhydrazyl (DPPH), ferric reducing antioxidant power (FRAP), and 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) methods, the antioxidant activities of fermented WB were found to be significantly improved. Also, there was a good correlation between the total phenolics content (TPC) and antioxidant capacity. Briefly, fermentation not only provides a new avenue to increase the functional potential of WB, but also enhances the utilization of agricultural byproducts.

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Site-directed mutagenesis of coenzyme-independent carotenoid oxygenase CSO2 to enhance the enzymatic synthesis of vanillin

Cited by 20 publications

References 28 publications

Protein Design: From the Aspect of Water Solubility and Stability

Protein Design: From the Aspect of Water Solubility and Stability

Whole-Cell Bioconversion of Renewable Biomasses-Related Aromatics to cis,cis-Muconic Acid

Process Optimization for Production of Ferulic Acid and Pentosans from Wheat Brans by Solid-State Fermentation and Evaluation of Their Antioxidant Activities

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