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Optimisation of Hydrocortisone Production by Curvularia lunata

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Abstract

A new method for breeding the hydrocortisone overproducing strain Curvularia lunata by screening ketoconazole-resistance mutant was developed. A hydrocortisone overproducing mutant C. lunata KA-91 with ketoconazole-resistance marker was obtained from protoplasts treated with ultraviolet radiation. The hydrocortisone conversion rate of C. lunata KA-91 was increased by 42.1% compared to the original strain CL-114 at the substrate 17α-hydroxypregn-4-en-3, 20-dione-21-acetate addition concentration of 1.0 g/L. The by-products produced by KA-91 were fewer than those of the original strain. It was assumed that the higher cytochrome P450 content of ketoconazole-resistance mutant resulted in the increase of 11β-hydroxylation capacity. The culture conditions for biotransformation of 17α-hydroxypregn-4-en-3, 20-dione-21-acetate to hydrocortisone were optimized by response surface methodology. Plackett–Burman design was applied to elucidate the key factors affecting the hydrocortisone production, and the results indicated that glucose, initial pH, and glucose to total nitrogen sources ratio (ω) had significant effects on hydrocortisone production. Box–Behnken design was employed to search for the optimal parameters of those three key factors. According to the model, the trial checking at the optimal conditions showed a high hydrocortisone conversion rate of 82.67%.

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Abbreviations

HCCR:

hydrocortisone conversion rate (%)

RT:

retain time (min)

Y :

experimental response of Plackett–Burman design (HCCR, %)

Z :

experimental response of Box–Behnken design (HCCR, %)

x i :

coded value of the variable X i

X i :

variable

X 0 :

value of X i at the center point

X :

step change

α i :

linear coefficient

α 0 :

intercept of α i

β i :

linear coefficients

β ii :

squared coefficients

β ij :

interaction coefficients

β 0 :

intercept of β i

ω :

glucose to total nitrogen sources ratio

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Acknowledgments

The authors wish to acknowledge the financial support provided by the Key Science and Technology Development Foundation of Tianjin (No. 023180211) and the Key Science and Technology Research Program of Chinese Educational Ministry (No. 01004).

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Authors and Affiliations

  1. School of Bioengineering, Tianjin University of Science and Technology, Tianjin, 300222, PR China

    Wenyu Lu, Lianxiang Du, Min Wang, Yawen Guo, Wei Gong, Huike Bao & Jing Yang

  2. School of Chemical Engineering and Technology, Tianjin University, Tianjin, 300072, PR China

    Wenyu Lu, Xiaoqiang Jia, Jianping Wen & Bing Sun

  3. School of Biological and Chemical Engineering, Tianjin University of Technology, Tianjin, 300191, PR China

    Yuping Huang

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  1. Wenyu Lu
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Correspondence to Lianxiang Du.

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Lu, W., Du, L., Wang, M. et al. Optimisation of Hydrocortisone Production by Curvularia lunata . Appl Biochem Biotechnol 142, 17–28 (2007). https://doi.org/10.1007/s12010-007-0005-8

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