Food and Fermentation Industries >

2025 , Vol. 51 >Issue 21: 27 - 34

DOI: https://doi.org/10.13995/j.cnki.11-1802/ts.042062

Biosynthesis of p-anisaldehyde from trans-anethole by whole-cell biotransformation using recombinant Escherichia coli

  • ZHANG Zhikai ,
  • LIN Qian
Expand
  • 1(College of Biology and Pharmacy, Yulin Normal University, Yulin 537000, China)
    2(Key Laboratory of Mountain Biodiversity Conservation, Education Department of Guangxi Zhuang Autonomous Region, Yulin Normal University, Yulin 537000, China)

Received date: 2025-01-08

  Revised date: 2025-02-19

  Online published: 2025-11-21

Fold

Abstract

p-Asnisaldehyde is an aromatic compound widely used in food, beverage, and cosmetics industries.To increase biosynthesis yield and conversion rate, whole-cell biocatalysis for p-anisaldehyde production was investigated.The trans-anethole oxygenase from Paraburkholderia sp.MR185 and a solubilization tag retro-protein XXA was fused and expressed in Escherichia coli BL21(DE3).The induced E.coli cells were used as whole-cell biocatalyst for conversion of trans-anethole to p-anisaldehyde, and were immobilized.The activity of trans-anethole oxygenase was dectected in induced E.coli cells and the fusion protein was present in the supernatant and sediment of cell extract.The induction conditions were optimized and the enzyme activity in E.coli cells reached 9.3 U/g.Under optimized conditions of whole-cell transformation, the yield of p-anisaldehyde reached 1.62 g/L with the conversion rate being 88.3%.E.coli cells were immobilized in sodium alginate and used for repeated transformation of trans-anethole to p-anisaldehyde.The yield of of p-anisaldehyde reached 1.26 g/L with the conversion rate being 68.5%, and in the fisrt 3 runs the yield of p-anisaldehyde were above 1 g/L.This study is the first report on whole-cell biocatalyst for p-anisaldehyde synthesis, and provides a technical reference for the industrialized biosynthesis of p-anisaldehyde.

Key words： p-anisaldehyde; trans-anethole; trans-anethole oxygenase; Escherichia coli; whole-cell biocatalysis

Cite this article

ZHANG Zhikai , LIN Qian . Biosynthesis of p-anisaldehyde from trans-anethole by whole-cell biotransformation using recombinant Escherichia coli[J]. Food and Fermentation Industries, 2025 , 51(21) : 27 -34 . DOI: 10.13995/j.cnki.11-1802/ts.042062

References

[1] XIONG J, YUAN X, ZONG M H, et al.Iron-incorporated metal-organic frameworks for oxidative cleavage of trans-anethole to p-anisaldehyde[J].Nanoscale, 2023, 15(48):19493-19498.
[2] MANINA A S, FORLANI F.Biotechnologies in perfume manufacturing:Metabolic engineering of terpenoid biosynthesis[J].International Journal of Molecular Sciences, 2023, 24(9):7874.
[3] DICKEY R M, GOPAL M R, NAIN P, et al.Recent developments in enzymatic and microbial biosynthesis of flavor and fragrance molecules[J].Journal of Biotechnology, 2024, 389:43-60.
[4] HAN D F, RYU J Y, KANALY R A, et al.Isolation of a gene responsible for the oxidation of trans-anethole to para-anisaldehyde by Pseudomonas putida JYR-1 and its expression in Escherichia coli[J].Applied and Environmental Microbiology, 2012, 78(15):5238-5246.
[5] HAN D F, SADOWSKY M J, CHONG Y, et al.Characterization of a self-sufficient trans-anethole oxygenase from Pseudomonas putida JYR-1[J].PLoS One, 2013, 8(9):e73350.
[6] 粟桂娇, 刘雄民, 朱萍, 等.反式茴脑生物降解菌株的筛选及其生物转化[J].化工进展, 2009, 28(1):141-144;172.
SU G J, LIU X M, ZHU P, et al.Isolation of trans-anethole degrading bacterial strains and biotransformation of trans-anethole[J].Chemical Industry and Engineering Progress, 2009, 28(1):141-144;172.
[7] 粟桂娇, 刘雄民, 李伟光, 等.两相系统中生物转化茴脑生成茴香醛[J].精细化工, 2009, 26(2):136-141.
SU G J, LIU X M, LI W G, et al.Biotransformation of anethole to anisaldehyde in aqueous-organic solvents biphasic systems[J].Fine Chemicals, 2009, 26(2):136-141.
[8] 梁肖仍, 宋张杨, 曾华贺, 等.一株转化茴脑生成茴香醛的菌株筛选与鉴定[J].食品与生物技术学报, 2011, 30(6):956-961.
LIANG X R, SONG Z Y, ZENG H H, et al.Isolation and characterization of a strain for bioconversion of anethole to anialdehyde[J].Journal of Food Science and Biotechnology, 2011, 30(6):956-961.
[9] 刘思琴, 郑璞, 陈鹏程, 等.合成洋茉莉醛重组菌株的构建与产物鉴定[J].食品与发酵工业, 2018, 44(6):1-7.
LIU S Q, ZHENG P, CHEN P C, et al.Construction of a recombinant strain for biosynthesis of heliotropin and its product identification[J].Food and Fermentation Industries, 2018, 44(6):1-7.
[10] WEN P, WU D, ZHENG P, et al.Highly efficient biosynthesis of heliotropin by engineered Escherichia coli coexpressing trans-anethole oxygenase and formate dehydrogenase[J].Journal of Agricultural and Food Chemistry, 2019, 67(51):14121-14128.
[11] LIN Q, LI J N, LING X R, et al.Cloning and expression of a novel trans-anethole oxygenase gene from Paraburkholderia sp.MR185[J].Journal of General and Applied Microbiology, 2022, 68(3):163-167.
[12] PURWANI N N, ROZEBOOM H J, WILLERS V P, et al.Discovery of a new class of bacterial heme-containing CC cleaving oxygenases[J].New Biotechnology, 2024, 83:82-90.
[13] TAVANTI M, HOSFORD J, LLOYD R C, et al.ATP regeneration by a single polyphosphate kinase powers multigram-scale aldehyde synthesis in vitro[J].Green Chemistry, 2021, 23(2):828-837.
[14] XIE X, WU P, HUANG X C, et al.Retro-protein XXA is a remarkable solubilizing fusion tag for inclusion bodies[J].Microbial Cell Factories, 2022, 21(1):51.
[15] YE X, LING X, WU M, et al.Improving soluble expression of SARS-CoV-2 spike priming protease TMPRSS2 with an artificial fusing protein[J].International Journal of Molecular Sciences, 2023, 24(13):10475.
[16] 敖龙, 朱玲玉, 庞欣, 等.人工蛋白XXA促进人源肿瘤抑素Tumstatin可溶性表达[J].食品与发酵工业, 2024, 50(12):1-8.
AO L, ZHU L Y, PANG X, et al.Artificial protein XXA promotes soluble expression of human-derived Tumstatin[J].Food and Fermentation Industries, 2024, 50(12):1-8.
[17] HEBDITCH M, CARBALLO-AMADOR M A, CHARONIS S, et al.Protein-Sol:A web tool for predicting protein solubility from sequence[J].Bioinformatics, 2017, 33(19):3098-3100.
[18] 邹树平, 颜海蔚, 胡忠策, 等.固定化重组大肠杆菌细胞催化合成(R)-环氧氯丙烷[J].现代化工, 2013, 33(7):55-59.
ZOU S P, YAN H W, HU Z C, et al.Synthesis of (R)-epichlorohydrin catalyzed by immobilized recombinant Escherichia coli cells[J].Mordern Chemical Industry, 2013, 33(7):55-59.
[19] 魏小平, 李建平, 蓝小月.分光光度法测定微量茴香醛的研究[J].食品科学, 2002, 23(1):101-103.
WEI X P, LI J P, LAN X Y.Spectrophotometric determination of micro-amount of anisaldehyde by condensation reaction with tba[J].Food Science, 2002, 23(1):101-103.
[20] 李沂馨, 毛银, 刘伟, 等.生物基1,6-己二醇的全细胞催化及其过程强化[J].食品与发酵工业, 2025, 51(5):8-14.
LI Y X, MAO Y, LIU W, et al.Whole-cell catalysis of biobased 1,6-hexanediol and its fermentation enhancement[J].Food and Fermentation Industries, 2025, 51(5):8-14.
[21] 付声亮, 邹诗瑶, 高娃, 等.重组大肠杆菌全细胞转化苹果酸合成丙酮酸[J].中国食品学报, 2024, 24(7):219-228.
FU S L, ZOU S Y, GAO W, et al.Biosynthesis of pyruvate from malic acid by whole-cell biotransformation using recombinant Escherichia coli[J].Journal of Chinese Institute of Food Science and Technology, 2024, 24(7):219-228.
[22] AUEZOVA L, NAJJAR A, KFOURY M, et al.Antibacterial activity of free or encapsulated selected phenylpropanoids against Escherichia coli and Staphylococcus epidermidis[J].Journal of Applied Microbiology, 2020, 128(3):710-720.
[23] LIN Y, HUANG R, SUN X X, et al.The p-Anisaldehyde/β-cyclodextrin inclusion complexes as a sustained release agent:Characterization, storage stability, antibacterial and antioxidant activity[J].Food Control, 2022, 132:108561.
[24] TAZON A W, AWWAD F, MEDDEB-MOUELHI F, et al.Biotechnological advances in vanillin production:From natural Vanilla to metabolic engineering platforms[J].BioChem, 2024, 4(4):323-349.
Options
Outlines

/

〈 〉

Powered by Beijing Magtech Co. Ltd,.