Enhancement of Schizochytrium DHA synthesis by plasma mutagenesis aided with malonic acid and zeocin screening
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Schizochytrium sp. accumulates valuable polyunsaturated fatty acid (PUFA), such as docosahexaenoic acid (DHA). In order to increase DHA synthesis in this microorganism, physical or chemical mutagenesis aided with powerful screening methods are still preferable, as its DHA synthetic pathway has not yet been clearly defined for gene manipulation. To breed this agglomerate microorganism of thick cell wall and rather large genome for increasing lipid content and DHA percentage, a novel strategy of atmospheric and room temperature plasma (ARTP) mutagenesis coupled with stepped malonic acid (MA) and zeocin resistance screening was developed. The final resulted mutant strain mz-17 was selected with 1.8-fold increased DHA production. Accompanied with supplementation of Fe2+ in shake flask cultivation, DHA production of 14.0 g/L on average was achieved. This work suggests that ARTP mutation combined with stepped MA and zeocin resistance screening is an efficient method of breeding Schizochytrium sp. of high DHA production, and might be applied on other microorganisms for obtaining higher desired PUFA products.
KeywordsSchizochytrium sp. Docosahexaenoic acid (DHA) Atmospheric and room temperature plasma (ARTP) Malonic acid (MA) Zeocin Fe·EDTA
This work was supported by the National High Technology Research and Development Program of China (No. 2014AA021702); the Program of the Innovation Plan of Jiangsu Province (No. KYLX_1148); the Fundamental Research Funds for the Central Universities (No. JUSRP51402A); the Program of the Key Laboratory of Industrial Biotechnology, Ministry of Education, China (No. KLIB-KF201601), and the 111 Project (No. 111-2-06).
Compliance with ethical standards
Conflict of interest
The authors declare that they have no competing interests.
This paper does not contain any studies with human participants or animals performed by any of the authors.
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