Construction and Optimization of an Efficient Tyrosol Biosynthesis System in Escherichia coli

Authors

Yizhe XIONG, School of Biotechnology , Jiangnan University , Wuxi 214122 , China ;Key Laboratory of Industrial Biotechnology of Ministry of Education , Jiangnan University , Wuxi 214122 , China
Shougang WANG, School of Biotechnology , Jiangnan University , Wuxi 214122 , China ;Key Laboratory of Industrial Biotechnology of Ministry of Education , Jiangnan University , Wuxi 214122 , China
Zhiming RAO, School of Biotechnology , Jiangnan University , Wuxi 214122 , China ;Key Laboratory of Industrial Biotechnology of Ministry of Education , Jiangnan University , Wuxi 214122 , China
Meijuan XU, School of Biotechnology , Jiangnan University , Wuxi 214122 , China ;Key Laboratory of Industrial Biotechnology of Ministry of Education , Jiangnan University , Wuxi 214122 , ChinaFollow

Corresponding Author(s)

徐美娟（1982—），女，博士，教授，博士研究生导师，主要从事微生物发酵菌种的系统代谢工程选育、合成生物学及关键酶改造方面的研究。E-mail：xumeijuan@jiangnan.edu.cn

Abstract

[Objective ] An efficient biosynthetic system was constructed in Escherichia coli to achieve high-yield production of tyramine and its key derivative tyrosol.[Method ] Through the analysis of protein expression,enzymatic properties,and whole-cell catalysis results,tyrosine decarboxylase (TDC ) derived from Enterococcus faecalis was selected as the key enzyme for tyramine synthesis.Subsequently,tyramine oxidase (TYO ) was introduced to construct the TDC-TYO pathway for tyrosol production.Meanwhile,the LAAD-PDC pathway for tyrosol synthesis was established.A dual-enzyme co-expression system was established,and the effects of tandem enzyme order,production pathways,and key enzyme sources on tyrosol yield were analyzed.The recombinant strain B 1,carrying the plasmid pET 28a-EfTDC-TYO,was obtained through whole-cell catalysis,and the reaction conditions were optimized.[Result ] Under optimal reaction parameters,the recombinant strain B 1 achieved a tyrosol yield of 3.96 g/L within 24 h,with a substrate conversion rate of 51.9%.[Conclusion ] This study provides valuable insights for the biosynthesis of tyrosol and its derivatives.

Publication Date

12-15-2025

First Page

18

Last Page

29

DOI

10.12441/spyswjs.20250312001

Recommended Citation

XIONG, Yizhe; WANG, Shougang; RAO, Zhiming; and XU, Meijuan (2025) "Construction and Optimization of an Efficient Tyrosol Biosynthesis System in Escherichia coli," Journal of Food Science and Biotechnology: Vol. 44: Iss. 12, Article 3.
DOI: 10.12441/spyswjs.20250312001
Available at: https://spsw.spyswjs.cnjournals.com/journal/vol44/iss12/3

References

［1］ KARKOVIĆ MARKOVIĆ A，TORIĆ J，BARBARIĆ M，et al.Hydroxytyrosol，tyrosol and derivatives and their potential effects on human health ［J］.Molecules，2019，24（10）：2001.
［2］ GABBIA D.Beneficial effects of tyrosol and oleocanthal from extra virgin olive oil on liver health：insights into their mechanisms of action ［J］.Biology，2024，13（10）：760.
［3］ KOUKA P，TSAKIRI G，TZORTZI D，et al.The polyphenolic composition of extracts derived from different Greek extra virgin olive oils is correlated with their antioxidant potency ［J］.Oxidative Medicine and Cellular Longevity，2019，2019：1870965.
［4］ PLOTNIKOV M B，PLOTNIKOVA T M.Tyrosol as a neuroprotector：strong effects of a “weak” antioxidant ［J］.Current Neuropharmacology，2021，19（4）：434-448.
［5］ ČERNÁKOVÁ L，MACKOVÁ M，KLEMPOVÁ T，et al.Enzymatic methoxycarbonylation of tyrosol and hydroxytyrosol ［J］.International Journal of Molecular Sciences，2024，25（18）：10057.
［6］ XIONG T Z，LI X M，LIU W，et al.Multienzyme cascade for synthesis of hydroxytyrosol via engineered Escherichia coli［J］.Scientific Reports，2025，15：471.
［7］ VAN KESSEL S P，FRYE A K，EL-GENDY A O，et al.Gut bacterial tyrosine decarboxylases restrict levels of levodopa in the treatment of Parkinson ’s disease［J］.Nature Communications，2019，10：310.
［8］ 刘英杰，符长春，张学鹏，等.酪醇及其衍生物的微生物代谢工程调控研究进展 ［J］.生物工程学报，2024，40（8）：2604 -2625.LIU Y J，FU C C，ZHANG X P，et al.Recent advances in metabolic engineering of microorganisms for production of tyrosol and its derivatives ［J］.Chinese Journal of Biotechnology，2024，40（8）：2604 -2625.（in Chinese ）
［9］ ZENG W Z，WANG H J，CHEN J B，et al.Engineering Escherichia coli for efficient de novo synthesis of s alidroside［J］.Journal of Agricultural and Food Chemistry，2024，72（51）：28369 -28377.
［10］ LIU H Y，TIAN Y J，ZHOU Y，et al.Multi-modular engineering of Saccharomyces cerevisiae for high-titre production of tyrosol and salidroside ［J］.Microbial Biotechnology，2021，14（6）：2605 -2616.
［11］ HAZELWOOD L A，DARAN J M，VAN MARIS A J A，et al.The Ehrlich pathway for fusel alcohol production：a century of research on Saccharomyces cerevisiae metabolism ［J］.Applied and Environmental Microbiology，2008，74（8）：2259 -2266.
［12］ SATOH Y，TAJIMA K，MUNEKATA M，et al.Engineering of a tyrosol-producing pathway，utilizing simple sugar and the central metabolic tyrosine，in Escherichia coli ［J］.Journal of Agricultural and Food Chemistry，2012，60（4）：979-984.
［13］ XIA Y Y，QI L N，SHI X L，et al.Construction of an Escherichia coli cell factory for de novo synthesis of tyrosol through semi-rational design based on phenylpyruvate decarboxylase ARO 10 engineering ［J］.International Journal of Biological Macromolecules，2023，253：127385.
［14］ CHUNG D，KIM S Y，AHN J H.Production of three phenylethanoids，tyrosol，hydroxytyrosol，and salidroside，using plant genes expressing in Escherichia coli ［J］.Scientific Reports，2017，7：2578.
［15］ LIU J B，WANG K P，WANG M，et al.Efficient whole cell biotransformation of tyrosol from L-tyrosine by engineered Escherichia coli ［J］.Enzyme and Microbial Technology，2022，160：110100.
［16］ RUAN X B，ZHANG S，SONG W，et al.Efficient synthesis of tyrosol from L-tyrosine via heterologous Ehrlich pathway in Escherichia coli ［J］.Chinese Journal of Chemical Engineering，2022，47：18-30.
［17］ 张月，裴慧洁，何维，等.发酵食品中酪胺的形成机制及其控制研究进展 ［J］.食品科学，2024，45（15）：263-271.ZHANG Y，PEI H J，HE W，et al.Research progress on the formation mechanism and control of tyramine in fermented foods ［J］.Food Science，2024，45（15）：263-271.（in Chinese ）
［18］ LIU J，LIU J M，YANG B，et al.Production of phenylpyruvic acid by engineered l-amino acid deaminase from Proteus mirabilis ［J］.Biotechnology Letters，2022，44（5）：635-642.
［19］ SUTIONO S，CARSTEN J，SIEBER V.Structure-guided engineering of α-keto acid decarboxylase for the production of higher alcohols at elevated temperature ［J］.ChemSusChem，2018，11（18）：3335 -3344.
［20］ CROWE J，MASONE B S，RIBBE J.One-step purification of recombinant proteins with the 6xHis tag and Ni-NTA resin ［J］.Molecular Biotechnology，1995，4（3）：247-258.
［21］ PEREZ M，LADERO V，DEL RIO B，et al.The relationship among tyrosine decarboxylase and agmatine deiminase pathways in Enterococcus faecalis ［J］.Frontiers in Microbiology，2017，8：2107.
［22］ 张慧，曹慧，王瑾，等.高效液相色谱 -三重四极杆串联质谱法分析奶酪中 5种生物胺 ［J］.食品与机械，2023，39（10）：80-86.ZHANG H，CAO H，WANG J，et al.Determination of 5 kinds of biogenic amines in cheeses by high performance liquid chromatography-tandem mass spectrometry ［J］.Food & Machinery，2023，39（10）：80-86.（in Chinese ）
［23］ ANDERSEN G，MARCINEK P，SULZINGER N，et al.Food sources and biomolecular targets of tyramine ［J］.Nutrition Reviews，2019，77（2）：107-115.
［24］ LUCAS P，LANDETE J，COTON M，et al.The tyrosine decarboxylase operon of Lactobacillus brevis IOEB 9809：characterization and conservation in tyramine-producing bacteria［J］.FEMS Microbiology Letters，2003，229（1）：65-71.
［25］ KANG H R，KIM H S，MAH J H，et al.Tyramine reduction by tyrosine decarboxylase inhibitor in Enterococcus faecium for tyramine controlled cheonggukjang ［J］.Food Science and Biotechnology，2018，27（1）：87-93.
［26］ 殷方荣，翟李欣，田乔鹏，等.贝莱斯芽孢杆菌角蛋白酶的克隆表达、纯化及酶学性质 ［J］.食品与生物技术学报，2023，42（6）：57-66.YIN F R，ZHAI L X，TIAN Q P，et al.Gene cloning，expression，purification，and enzymatic properties study of keratinase from Bacillus velezensis ［J］.Journal of Food Science and Biotechnology，2023，42（6）：57-66.（in Chinese）
［27］ YU T Q，YIN Y F，GE Y H，et al.Enzymatic production of 4-hydroxyphenylacetaldehyde by oxidation of the amino group of tyramine with a recombinant primary amine oxidase ［J］.Process Biochemistry，2020，92：105-112.
［28］ SHEN N，SATOH Y，KOMA D，et al.Optimization of tyrosol-producing pathway with tyrosine decarboxylase and tyramine oxidase in high-tyrosine-producing Escherichia coli ［J］.Journal of Bioscience and Bioengineering，2024，137（2）：115-123.
［29］ 郑蝶，杨海泉，夏媛媛，等.重组大肠杆菌全细胞催化法生产 N-乙酰神经氨酸 ［J］.食品与生物技术学报，2024，43（2）：38-46.ZHENG D，YANG H Q，XIA Y Y，et al.Production of N-acetylneuraminic acid from fermentation broth using recombinant Escherichia coli whole cells biotransformation ［J］.Journal of Food Science and Biotechnology，2024，43（2）：38-46.（in Chinese ）
［30］ KNAPP B D，HUANG K C.The effects of temperature on cellular physiology ［J］.Annual Review of Biophysics，2022，51：499-526.
［31］ GUO J，MA Z P，GAO J S，et al.Recent advances of pH homeostasis mechanisms in Corynebacterium glutamicum［J］.World Journal of Microbiology and Biotechnology，2019，35（12）：192.
［32］ LI X B，LI K H，FARAJTABAR A，et al.Solubility of d-tryptophan and l-tyrosine in several organic solvents：determination and solvent effect ［J］.Journal of Chemical & Engineering Data，2019，64（7）：3164 -3169.
［33］ WANG J L，MA W J，ZHOU J W，et al.Microbial chassis design and engineering for production of amino acids used in food industry ［J］.Systems Microbiology and Biomanufacturing，2023，3（1）：28-48.