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. 2025 Apr;197(4):2579-2597.
doi: 10.1007/s12010-024-05145-5. Epub 2025 Jan 8.

Effect of Fold-Promoting Mutation and Signal Peptide Screening on Recombinant Glucan 1,4-Alpha-maltohydrolase Secretion in Pichia pastoris

Siyi Wang  1 , Kai Zhu  1 , Pulin Liu  2
Affiliations

Effect of Fold-Promoting Mutation and Signal Peptide Screening on Recombinant Glucan 1,4-Alpha-maltohydrolase Secretion in Pichia pastoris

Siyi Wang et al. Appl Biochem Biotechnol. 2025 Apr.

Abstract

Glucan 1,4-alpha-maltohydrolase (3.2.1.133, GMH) is an important biocatalyst in the baking industry, which could delay the retrogradation of bread and improve its cold-storage durability. In the present study, a newly cloned Thgmh was characterized and secreted by Pichia pastoris (Komagataella pastoris). After computationally assisted rational design that promotes peptide folding, the maltogenic activity in supernatant was enhanced 1.6-fold in comparison with the base strain. The signal leading sequence screening and the gene dosage increment further improved secretion by approximately 6.4-fold. The purified rationally designed ThGMHs exhibited maximal activity against soluble starch at pH 7.0 and 60 ℃, and maltose is the main catalytic product. In a 5-L bioreactor, conventional fed-batch fermentation resulted in 6130 U mL-1 extracellular maltogenic activity. Therefore, a promising strain for GMH production was developed, which provides a useful reference for the secretory production of other industrial enzymes.

Keywords: Pichia pastoris; Fermentation; Glucan 1,4-alpha-maltohydrolase; Rational design; Signal leading sequence.

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Conflict of interest statement

Declarations. Ethics Approval: This study does not contain any experiments involving human or animal subjects. Consent to Participate: Informed consent was obtained from all individual participants included in this study. Consent to Publish: The authors agreed to publish this paper in Applied Biochemistry and Biotechnology. Competing Interests: The authors declare no competing interests.

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References

    1. Miao, M., & BeMiller, J. N. (2023). Enzymatic approaches for structuring starch to improve functionality. Annual Review of Food Science and Technology, 14, 271–295. https://doi.org/10.1146/annurev-food-072122-023510 - DOI - PubMed
    1. Srichuwong, S., & Jane, J. L. (2007). Physicochemical properties of starch affected by molecular composition and structures: A review. Food Science and Biotechnology, 16, 663–674.
    1. Miao, M., Jiang, B., Jin, Z. Y., & BeMiller, J. N. (2018). Microbial starch-converting enzymes: Recent insights and perspectives. Comprehensive Reviews in Food Science and Food Safety, 17, 1238–1260. https://doi.org/10.1111/1541-4337.12381 - DOI - PubMed
    1. BeMiller, J. N., & Huber, K. C. (2015). Physical modification of food starch functionalities. Annual Review of Food Science and Technology, 6, 19–69. https://doi.org/10.1146/annurev-food-022814-015552 - DOI - PubMed
    1. Bangar, S. P., Ashogbon, A. O., Singh, A., Chaudhary, V., & Whiteside, W. S. (2022). Enzymatic modification of starch: A green approach for starch applications. Carbohydrate Polymers, 287, 119265. https://doi.org/10.1016/j.carbpol.2022.119265 - DOI

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