Biosynthesis of 3-（4-Hydroxyphenyl）Propionic Acid in Saccharomyces cerevisiae

doi:10.13560/j.cnki.biotech.bull.1985.2017-0208

Abstract

Abstract: 3-（4-Hydroxyphenyl）propionic acid（HPPA）is an aromatic compound with importantly economic value,because it is an intermediate mainly used in medicine,food and chemical industry. In this work,we aim to achieve the De novo synthesis of HPPA in Saccharomyces cerevisiae,and it was conducted via overexpressing a tyrosine ammonia-lyase from Flavobacterium johnsoniaeu（FjTAL）and a p-coumaroyl-CoA ligase from Arabidopsis thaliana（At4CL）,as well as using super long-chain enoyl-CoA reductase（ScTSC13）and acyl-CoA thioesterases from S. cerevisiae BY4742. The yield of HPPA reached 70 mg/L. Bioconversion rate of tyrosine into p-coumaric acid was 36% under the condition of 4 mmol/L tyrosine enhancement in shake flask after 72 h fermentation at 30℃ and galactose inducing FjTAL overexpression. While in the recombinant S. cerevisiae overexpressing At4CL combined with enzymes ScTSC13 and acyl-CoA thioesterases,94% of 4 mmol/L p-coumaric acid was converted to HPPA in shake flask after 72 h fermentation at 30℃,and 91 % of 4 mM caffeic acid was converted to DHPPA. This work laid a foundation for the biosynthesis of HPPA and its derivatives.

Key words: Saccharomyces cerevisiae, 3-（4-hydroxyphenyl）propionic acid, biosynthesis, tyrosine ammonia-lyase, p-coumaroyl-coA ligase

JIANG Jing-jie,ZHUANG Yi-bin,YIN Hua,LIU Tao,LIU Shao-wei. Biosynthesis of 3-（4-Hydroxyphenyl）Propionic Acid in Saccharomyces cerevisiae[J]. Biotechnology Bulletin, 2017, 33(9): 184-190.

References

[1] Araki H, Kawabata A, Kuroda R, et al. Compositions for preventing and treating digestive organs diseases：US, US7550437[P] . 2009.
[2] Tang YH, Wang JY, Hu HH, et al. Analysis of species-dependent hydrolysis and protein binding of esmolol enantiomers[J] . Journal of Pharmaceutical Analysis, 2012, 2（3）：220-225.
[3] Kawai S, Nakata K, Ichizawa H, et al. 3-（4-Hydroxyphenyl）propionic acid is involved in the biosynthesis of myricanol in Myrica rubra[J] . Journal of Wood Science, 2012, 56（2）：148-153.
[4] Yahyaa M, Davidovich-Rikanati R, Eyal Y, et al. Identification and characterization of UDP-glucose：Phloretin 4'-O-glycosyltransferase from Malus x domestica Borkh[J] . Phytochemistry, 2016, 130：47-55.
[5] Croitoru R, Fi?ig?u F, Broek LAMVD, et al. Biocatalytic acylation of sugar alcohols by 3-（4-hydroxyphenyl）propionic acid[J] . Process Biochemistry, 2012, 47（12）：1894-1902.
[6] 管惠娟, 张雪, 屠凤娟, 等. 铁皮石斛化学成分的研究[J] . 中草药, 2009, 40（12）：1873-1876.
[7] 黄洋, 邵慧凯, 李康, 等. 小叶榕叶抗炎成分分析及活性评价[J] . 中成药, 2014, 36（6）：1227-1233.
[8] 张明星, 盛喆. 对羟基苯丙酸的制备与应用[J] . 精细与专用化学品, 2010, 18（9）：48-49.
[9] 李晓林, 周威, 庄以彬, 等. 3, 4-二羟基扁桃酸在大肠杆菌中的生物合成[J] . 生物技术通报, 2017, 33（1）：135-140.
[10] Jing S, Lin Y, Shen X, et al. Aerobic biosynthesis of hydrocinnamic acids in Escherichia coli, with a strictly oxygen-sensitive enoate reductase[J] . Metabolic Engineering, 2016, 35：75-82.
[11] Hong KK, Nielsen J. Metabolic engineering of Saccharomyces cerevisiae：a key cell factory platform for future biorefneries[J] . Cellular & Molecular Life Sciences Cmls, 2012, 69（16）：2671-2690.
[12] Gosch C, Halbwirth H, Stich K. Phloridzin：biosynthesis, distribution and physiological relevance in plants[J] . Phytochemistry, 2010, 71（8-9）：838-843.
[13] Jiang J, Bi H, Zhuang Y, et al. Engineered synthesis of rosmarinic acid in Escherichia coli, resulting production of a new intermediate, caffeoyl-phenyllactate[J] . Biotechnology Letters, 2016, 38（1）：81-88.
[14] Jendresen CB, Stahlhut SG, Li M, et al. Novel highly active and specific tyrosine ammonia-lyases from diverse origins enable enhanced production of aromatic compounds in bacteria and yeast[J] . Applied & Environmental Microbiology, 2015, 81（13）：4458-76.
[15] Rodriguez A, Kildegaard KR, Li M, et al. Establishment of a yeast platform strain for production of p- coumaric acid through metabolic engineering of aromatic amino acid biosynthesis[J] . Metabolic Engineering, 2015, 31：181.
[16] Sepp D. Kohlwein, Sandra E, et al. Tsc13p is required for fatty acid elongation and localizes to a novel structure at the nuclear-vacuolar interface in Saccharomyces cerevisiae[J] . 2001, 21（1）：109-125.
[17] Eichenberger M, Lehka BJ, Folly C, et al. Metabolic engineering of Saccharomyces cerevisiae for de novo production of dihydrochalc-ones with known antioxidant, antidiabetic, and sweet tasting prope-rties[J] . Metabolic Engineering, 2016.
[18] Gietz RD, Woods RA. Transformation of yeast by lithium acetate/single-stranded carrier DNA/polyethylene glycol method[J] . Methods in enzymology, 2002, 350（350）：87-96.
[19] Beekwilder J, Wolswinkel R, Jonker H, et al. Production of resveratrol in recombinant microorganisms[J] . Applied & Environmental Microbiology, 2006, 72（8）：5670-5672.
[20] Kal AJ, Hettema EH, Van dBM, et al. In silicio search for genes encoding peroxisomal proteins in Saccharomyces cerevisiae[J] . Cell biochemistry and biophysics, 2000, 32（1）：1-8.
[21] Maeda I, Delessert S, Hasegawa S, et al. The peroxisomal Acyl-CoA thioesterase Pte1p from Saccharomyces cerevisiae is required for efficient degradation of short straight chain and branched chain fatty acids[J] . Journal of Biological Chemistry, 2006, 281（17）：11729-11735.