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Cloning and functional analysis of putative malonyl-CoA:acyl-carrier protein transacylase gene from the docosahexaenoic acid-producer Schizochytrium sp. TIO1101

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Abstract

Malonyl-CoA:acyl-carrier protein transacylase (MCAT), which transfers the malonyl group from malonyl-CoA to holo-acyl carrier protein (ACP), is a key enzyme in fatty acid biosynthesis. Schizochytrium sp. TIO1101 is a marine protist with high levels of docosahexaenoic acid accumulation. In this study, the putative fabD gene coding MCAT was isolated from Schizochytrium sp. TIO1101. The Schizochytrium MCAT gene (ScTIOfabD) contained an 1176 bp open reading frame encoding a protein of 391 amino acids. The ScTIOfabD gene exhibited high novelty in nucleotide and amino acid sequence. The highest amino acid identity was only 35 % between ScTIOMCAT and the reported MCATs. Further studies demonstrated that ScTIOMCAT could bind malonyl-CoA directly and transfer malonyl group from malonyl-CoA to the ACP domain in vitro. Phylogenetic analysis suggested that ScTIOMCAT was relative close to MCATs of yeast strains. Overexpression of ScTIOMCAT in Saccharomyces cereviseae significantly increased the MCAT activity, without negative effects on the growth rate of the host strain. In addition, ScTIOMCAT generated 16.8 and 62 % increase in biomass and fatty acid accumulation, respectively, and did not alter the profile of fatty acid. Our results indicated that the novel MCAT gene from Schizochytrium sp. TIO1101 was crucial for fatty acid synthesis and had potential applications for genetic modifications of oil-producing species.

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Acknowledgments

This work was supported by the National Natural Science Foundation of China (No. 81202820), Zhejiang Provincial Natural Science Foundation of China (Grant No. LQ12H28005), Zhejiang Provincial Administration of Traditional Chinese Medicine (No. 2012ZQ006) and Scientific Research Fund of Zhejiang Provincial Education Department (No. Y201223804).

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Authors and Affiliations

  1. College of Pharmaceutical Science, Zhejiang Chinese Medical University, 548 Binwen Road, Hangzhou, 310003, China

    Rubin Cheng, Bo Yang, Xiaoming Zhong & Zhen Huang

  2. Zhejiang Hospital of Traditional Chinese Medicine, Zhejiang Chinese Medical University, 54 Youdian Road, Hangzhou, 310006, China

    Yuqing Ge

  3. Engineering Research Center of Marine Biological Resource Comprehensive Utilization, Third Institute of Oceanography, State Oceanic Administration, 178 Daxue Road, Xiamen, 361005, China

    Xiangzhi Lin

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  1. Rubin Cheng
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  2. Yuqing Ge
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  3. Bo Yang
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  4. Xiaoming Zhong
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  5. Xiangzhi Lin
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  6. Zhen Huang
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Correspondence to Zhen Huang.

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Cheng, R., Ge, Y., Yang, B. et al. Cloning and functional analysis of putative malonyl-CoA:acyl-carrier protein transacylase gene from the docosahexaenoic acid-producer Schizochytrium sp. TIO1101. World J Microbiol Biotechnol 29, 959–967 (2013). https://doi.org/10.1007/s11274-013-1253-0

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