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One-Pot Purification and Immobilization of Phenylalanine Dehydrogenase from Bacillus nanhaiensi by Functional Reduced Graphene Oxide

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Abstract

The one-pot immobilization of halophilic phenylalanine dehydrogenase from marine microorganism with metal ions modified reduced graphene oxide (CRGO) material was studied. Phenylalanine dehydrogenase was from Bacillus nanhaiensi and expressed with a C-terminal His-tag. Investigation of CRGO, CRGO-PEI, CRCO-Mn, and CRGO-PEI-Mn for one-pot purification and immobilization of phenylalanine dehydrogenase from crude enzyme solution was carried out. Enzyme activity yield rate achieved 80.0% by immobilization with CRCO-Mn, and the loading capacity was 6.7 mg/mg. Manganese ion coordination greatly improved the selectivity of the CRGO for the target His-tagged enzyme. Furthermore, the effect of NaCl concentration on the immobilization was investigated, which the loading capacity of CRGO-PEI and CRGO-Mn-PEI was increased by 10.7% and 30.6% with 1 M NaCl, respectively. The adsorption curves of crude enzyme one-pot immobilized by CRGO-Mn and purified enzyme immobilized by CRGO-Mn were similar. Therefore, one-pot immobilization strategy is promising for industrial application with advantages such as high efficiency and low cost, which shorten the pipelines for enzyme discovery towards industrial applications through the establishing of marine enzyme collections.

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The data sets supporting the results of this article are included within the article and its additional files.

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Abbreviations

CRGO:

Reduced graphene oxide

PEI:

Polyethylenimine

CRGO-PEI:

Polyethylenimine-grafted reduced graphene oxide

CRGO-Mn:

Manganese ions coordinated reduced graphene oxide

CRGO-PEI-Mn:

Manganese ions coordinated polyethylenimine-grafted reduced graphene oxide

PheDH:

Phenylalanine dehydrogenase

GO:

Graphene oxide

L-Phe:

L-phenylalanine

NAD(P)H:

Triphosphopyridine nucleotide

IPTG:

Kanamycin and isopropyl-beta-D-thiogalactopyranoside

NAD+ :

Nicotinamide adenine dinucleotide

U:

One unit

PEI:

Polyethyleneimine

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Funding

This work was supported by the National Natural Science Foundation of China (No. 21776233, 22078273), Natural Science Foundation of Fujian Province of China (No. 2018J01013), the Fundamental Research Funds for the Central Universities (No. 20720200038).

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Authors and Affiliations

  1. Department of Chemical and Biochemical Engineering, College of Chemistry and Chemical Engineering, Xiamen University, Xiamen, 361005, China

    Zhehui Ji, Guangxiao Yao, Liang Jiang & Shizhen Wang

  2. The Key Lab for Synthetic Biotechnology of Xiamen City, Xiamen University, Xiamen, Fujian, 361005, People’s Republic of China

    Shizhen Wang

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  1. Zhehui Ji
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  2. Guangxiao Yao
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Contributions

Zhehui Ji: methodology, investigation, analysis, interpretation, and writing. Guangxiao Yao: methodology, investigation, and analysis. Liang Jiang: investigation, analysis. Shizhen Wang: conception, design, funding acquisition, supervision, analysis and interpretation, drafting the manuscript, reading and final approval of the version to be published.

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Correspondence to Shizhen Wang.

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We declare that we have no financial and personal relationships with other people or organizations that can inappropriately influence our work, there is no professional or other personal interest of any nature or kind in any product, service and/or company that could be construed as influencing the position presented in, or the review of, the manuscript entitled, “One Pot Purification and Immobilization of Amino Acid Dehydrogenase by Functional Reduced Graphene Oxide.”

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Ji, Z., Yao, G., Jiang, L. et al. One-Pot Purification and Immobilization of Phenylalanine Dehydrogenase from Bacillus nanhaiensi by Functional Reduced Graphene Oxide. Mar Biotechnol 24, 555–565 (2022). https://doi.org/10.1007/s10126-022-10123-1

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