Improvement of Thermostability of Cholesterol Oxidase From Streptomyces Sp. Sa-Coo by Random Mutagenesis

Improvement of Thermostability of Cholesterol Oxidase From Streptomyces Sp. Sa-Coo by Random Mutagenesis Publisher Pubmed

Fazaeli A¹ ; Fana SE^{2, 3} ; Golestani A² ; Aminian M²

Show Affiliations

1. Department of Clinical Biochemistry, School of Medicine, Ardabil University of Medical Sciences, Ardabil, Iran
2. Department of Clinical Biochemistry, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran
3. Student Scientific Research Center, Tehran University of Medical Sciences, Tehran, Iran

Source: Protein Expression and Purification Published:2022

Abstract

To enhance the thermal stability of Streptomyces Sp. SA-COO cholesterol oxidase, random mutagenesis was used. A random mutant library was generated using two types of error-prone PCR (single step and serial dilution) and two mutants (ChOA-M1 and ChOA-M2) with improved thermostability were obtained. The best mutant ChOA-M1 acquired three amino acid substitutions (G49T, W52K, and F62V) and improved thermostability (at 50 °C for 5 h) by 40% and increased the kcat/Km value by 23%. The optimum pH was desirably changed to encompass a broad range from alkali to acid and circular dichroism revealed no significant secondary structure changes in mutants against wild type. These findings indicated that random mutagenesis was an effective technique for optimizing cholesterol oxidase properties and make a foundation for practical applications of Cholesterol oxidase in clinical diagnosis and industrial fields. © 2021

2. Expression Optimization of Recombinant Cholesterol Oxidase in Escherichia Coli and Its Purification and Characterization, AMB Express (2018)

3. Expression Optimization, Purification, and Functional Characterization of Cholesterol Oxidase From Chromobacterium Sp. Ds1, PLoS ONE (2019)

Experts (# of related papers)

Mahdi Aminian (3)

Abolfazl Golestani (2)

Style	Citing Format
MLA	Fazaeli A, et al.. "Improvement of Thermostability of Cholesterol Oxidase From Streptomyces Sp. Sa-Coo by Random Mutagenesis." Protein Expression and Purification, vol. 191, no. , 2022, pp. -.
APA	Fazaeli A, Fana SE, Golestani A, Aminian M (2022). Improvement of Thermostability of Cholesterol Oxidase From Streptomyces Sp. Sa-Coo by Random Mutagenesis. Protein Expression and Purification, 191(), -.
Chicago	Fazaeli A, Fana SE, Golestani A, Aminian M. "Improvement of Thermostability of Cholesterol Oxidase From Streptomyces Sp. Sa-Coo by Random Mutagenesis." Protein Expression and Purification 191, no. (2022): -.
Harvard	Fazaeli A et al. (2022) 'Improvement of Thermostability of Cholesterol Oxidase From Streptomyces Sp. Sa-Coo by Random Mutagenesis', Protein Expression and Purification, 191(), pp. -.
Vancouver	Fazaeli A, Fana SE, Golestani A, Aminian M. Improvement of Thermostability of Cholesterol Oxidase From Streptomyces Sp. Sa-Coo by Random Mutagenesis. Protein Expression and Purification. 2022;191():-.
BibTex	@article{ author = {Fazaeli A and Fana SE and Golestani A and Aminian M}, title = {Improvement of Thermostability of Cholesterol Oxidase From Streptomyces Sp. Sa-Coo by Random Mutagenesis}, journal = {Protein Expression and Purification}, volume = {191}, number = {}, pages = {-}, year = {2022} }
RIS	TY - JOUR AU - Fazaeli A AU - Fana SE AU - Golestani A AU - Aminian M TI - Improvement of Thermostability of Cholesterol Oxidase From Streptomyces Sp. Sa-Coo by Random Mutagenesis JO - Protein Expression and Purification VL - 191 IS - SP - EP - PY - 2022 ER -

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