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Two-Stage Cultivation of Pseudomonas sp. F12 for the Production of Enzymes Converting dl-2-Amino-Δ2-thiazoline-4-carboxylic Acid to l-Cysteine

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Abstract

Pseudomonas sp. F12 isolated from soil could transform dl-2-amino-Δ2-thiazoline-4-carboxylic acid (DL-ATC) to l-cysteine. It could grow in minimal medium containing DL-ATC as the sole carbon and nitrogen source, and the apparent activity of l-cysteine synthesis (CS) achieved 122 U/mL in a 5-L bioreactor. Pseudomonas sp. F12 could utilize glucose as carbon source and ammonia as nitrogen source for growth, but no CS activity was formed. To reduce the cost of DL-ATC, the cultivation process was divided into a growth stage on glucose and ammonia and a production stage induced by DL-ATC. The excessive glucose led to the production of byproduct(s) which seriously inhibited cell growth and CS production. Ammonium was accumulated when DL-ATC was consumed, and ammonium did not inhibit CS activity formation until 60 mM. Based on the above features, fed-batch cultivation of the growth stage was developed by supplying glucose restrictively. The volumetric CS activity was enhanced more than two times that obtained under the initial conditions.

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Acknowledgments

This work was supported by the “863” Program of China, grant No. 2011AA02A203.

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  1. State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, 130 Meilong Road, Shanghai, 200237, China

    Cuili Fan, Zhimin Li & Qin Ye

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  1. Cuili Fan
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  2. Zhimin Li
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Correspondence to Zhimin Li or Qin Ye.

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Fan, C., Li, Z. & Ye, Q. Two-Stage Cultivation of Pseudomonas sp. F12 for the Production of Enzymes Converting dl-2-Amino-Δ2-thiazoline-4-carboxylic Acid to l-Cysteine. Appl Biochem Biotechnol 168, 1867–1879 (2012). https://doi.org/10.1007/s12010-012-9903-5

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  • DOI: https://doi.org/10.1007/s12010-012-9903-5

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