The Effect on Phosphatidylserine Biosynthesis of Gene sdaA，sdaB，pgpB in Escherichia coli

Abstract

Abstract: The gene sdaA, sdaB, pgpB were deleted in Escherichia coli by the technology of Red homologous recombination system, resulting in the cut-off of part of the L-serine and PGP catabolic pathway, then the levels of phosphatidylserine（PS）could be elevated. To assess the effects of sdaA, sdaB, pgpB deletion on PS accumulation, both the recombinant and recipient strains were cultivated in LB-containing flasks. Then the samples were taken after 1 000 minutes and quantification of PS performed with HPLC. The results showed that the recombinant E. coli cells produced PS one fold more than the wild-type strain, suggesting the effectiveness of targeted alteration of the metabolic pathways of PS for improvement of PS production. Cost-effective large-scale production of PS should find important applications in various industrial fields.

Key words: Escherichia coli, Gene knockout, Phosphatidylserine, Red homologous recombination system, HPLC

Tong Xinwei, Yang Hongzhe, Li Yu, Lu Fuping . The Effect on Phosphatidylserine Biosynthesis of Gene sdaA，sdaB，pgpB in Escherichia coli[J]. Biotechnology Bulletin, 2013, 0(4): 123-128.

References

［1］ Kidd PM. Phosphatidylserine ：membrane nutrient for memory.
Aclinical and mechanistic assessment [J]. Alternative Medicine Review, 1996, 1（2）：70-84.
［2］ Su HS, Moniakis J, Newman EB. Use of gene fusions of the structural gene sdaA to purify L-serine deaminase 1 from Escherichia coli K-12 [J]. Eur J Biochem, 1993, 211（3）：521-527.
［3］ Su HS, Lang BF, Newman EB. L-Serine degradation in Escherichia coli K-12 ：Cloning and sequencing of the sdaA gene [J]. Journal of Bacteriology, 1989, 171（9）：5095-5102.
［4］ Shao ZQ, Newman EB. Sequencing and characterization of the sdaB gene from Escherichia coli K-12 [J]. Eur J Biochem, 1993, 212（3）: 777-784.
［5］ Icho T. Membrane-bound phosphatases in Escherichia coli：sequence of the pgpA gene [J]. Journal of Bacteriology, 1988, 170（11）: 5110-5116.
［6］ Funk CR, Zimniak L, Dowhan W. The pgpA and pgpB genes of Escherichia coli are not essential ：Evidence for a third phosphatidylglycerophosphate phosphatase [J]. Journal of Bacteriology, 1992, 174 （1）：205-213.
［7］ Ishinaga M, Kito M. Participation of soluble phosphatidylserine synthetasein pliosphatidylethanolamine biosynthesis in Escherichia coli membrane [J]. Eur J Biochem, 1974, 42 ：483-487.
［8］ Hawrott E, Kennedy EP. Phospholipid composition and membrane function in phosphatidylserine decarbo xylase mutants of Escherichia coli [J]. The Journal of Biological, 1978, 253（22）：8213-8220.
［9］ Yamamoto S, Izumiya H, Morita M, et al. Application of λRed recombination system to Vibrio cholerae genetics ：Simple methods for inactivation and modification of chromosomal genes [J]. Gene, 2009, 438（1-2）：57-64.
［10］ Maresca M, Erler A, Fu J, et al. Single-stranded heteroduplex intermediates in lambda Red homologous recombination [J]. BMC Mol Biol, 2010, 29 ：11-54.
［11］ Tischer BK, Smith GA, Osterrieder N. En passant mutagenesis : a two step markerless red recombination system [J]. Methods Mol Biol, 2010, 634 ：421-430.
［12］ Li Y, Chen GK, Tong XW, et al. Construction of Escherichia coli strains producing L-serine from glucose[J]. Biotechnol Lett, 2012, 34（8）：1525-1530.
［13］ Romański KW. The role and mechanism of action of bile acids in the Digestive System-Bile Acids in the gut [J]. Adv Clin Exp Med, 2008, 17（1）：83-89.
［14］ Russell DW. 50 years of advances in bile acid synthesis and metabolism [J]. Lipid Res ASBMB, 2009, 50（4）：120-125.
［15］ Blois A, Holmsen H, Martino G, et al. Interactions of chromogranin A-derived vasostatins and monolyers of phosphatidylserine, phosphatidylcholine and phosphatidylethanolamine [J].
Regul Pept, 2006, 134（1）：30-37.
［16］ Golovastov VV, Zolov SN, Nesmeyanova MA. Study of interaction of export initiation domain of Escherichia coli mature alkaline phosphatase with membrane phospholipids during secretion [J].
Biochemistry（Moscow）, 2002, 67（9）：978-985.

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