Skip to main content
SpringerLink
Log in

Regioselective O-acetylation of various glucosides catalyzed by Escherichia coli maltose O-acetyltransferase

  • Biotechnological Products and Process Engineering
  • Published:
Applied Microbiology and Biotechnology Aims and scope Submit manuscript

Abstract

Escherichia coli, a well-known prokaryotic organism, has been widely employed as a versatile host for heterologous overexpression of proteins/biocatalysts and the production of pharmaceutically important intermediates/small molecules. However, some E. coli endogenous enzymes showing substrate promiscuity may disturb the heterologous metabolic flux, which will result in the reduction of substrates, intermediates, and target products. Here we reported an unexpected E. coli-catalyzed regioselective O-acetylation of various glucosides. The regioselectively O-acetylated products, 6′-O-acetyl-loganin and 6′-O-acetyl-loganic acid, were obtained and characterized from the enzymatic reaction in which the supernatants of E. coli expressing either CaCYP72A565 and CaCPR, the key enzymes involved in camptothecin biosynthesis, or empty vector were used as catalyst and loganin and loganic acid as independent substrate. An alkaloidal glucoside strictosamide was converted into the regioselectively O-acetylated product 6′-O-acetyl-strictosamide, implying substrate promiscuity of the E. coli-catalyzed O-acetylation reaction. Furthermore, 8 glucosides, including 5 iridoid glucosides and 3 flavonoid glucosides, were successfully converted into the regioselectively O-acetylated products by E. coli, indicating the wide substrate range for the unexpected E. coli-catalyzed O-acetylation. E. coli maltose O-acetyltransferase was demonstrated to be responsible for the mentioned regioselective O-acetylation at the 6-OH of the glucopyranosyl group of multiple classes of natural product glucosides through candidate acetyltransferase-encoding gene analysis, gene knock-out, gene complementation, and the relevant enzymatic reaction activity assays. The present study not only provides an efficient biocatalyst for regioselective O-acetylation but also notifies cautions for metabolic engineering and synthetic biology applications in E. coli.

Graphical abstract

Key points

• 6-OH of glucosyl of multiple glucosides was regioselectively O-acetylated by E. coli.

• Endogenous EcMAT is responsible for the regioselective O-acetylation reaction.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

Data availability

The authors declare that the data supporting the findings of this study are available within the paper and its Supplementary Data files. Should any raw data files be needed in another format they are available from the corresponding author upon reasonable request.

References

  • Barnes HJ, Arlotto MP, Waterman MR (1991) Expression and enzymatic activity of recombinant cytochrome P450 17α-hydroxylase in Escherichia coli. Proc Natl Acad Sci U S A 88:5597–5601

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Brand B, Boos W (1991) Maltose transacetylase of Escherichia coli. J Biol Chem 266:14113–14118

    Article  CAS  PubMed  Google Scholar 

  • Cheng J, Luo Q, Duan H, Peng H, Zhang Y, Hu J, Lu Y (2020) Efficient whole-cell catalysis for 5-aminovalerate production from L-lysine by using engineered Escherichia coli with ethanol pretreatment. Sci Rep 10:990

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Cherepanov PP, Wackernagel W (1995) Gene disruption in Escherichia coli: TcR and KmR cassettes with the option of Flp-catalyzed excision of the antibiotic-resistance determinant. Gene 158:9–14

    Article  CAS  PubMed  Google Scholar 

  • Cravens A, Payne J, Smolke CD (2019) Synthetic biology strategies for microbial biosynthesis of plant natural products. Nat Commun 10:2142

    Article  PubMed  PubMed Central  Google Scholar 

  • Datsenko KA, Wanner BL (2000) One-step inactivation of chromosomal genes in Escherichia coli K-12 using PCR products. Proc Natl Acad Sci U S A 97:6640–6645

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Ding Y, Zhang D, Liu S, Dai J (2012) Selective acetylation of puerarin by Rhodococcus sp. J Chin Pharm Sci 21:269–272

    CAS  Google Scholar 

  • Erdelmeier CAJ, Wright AD, Orjala J, Baumgartner B, Rali T, Sticher O (1991) New indole alkaloid glycosides from Nauclea orientalis. Planta Med 57:149–152

    Article  CAS  PubMed  Google Scholar 

  • Figueroa-Bossi N, Balbontín R, Bossi L (2023) Recombineering 101: making an in-frame deletion mutant. Cold Spring Harb Protoc 2023:prot107856

    Article  Google Scholar 

  • Firouzabadi FN, Vincken J, Ji Q, Suurs LCJM, Visser RGF (2007) Expression of an engineered granule-bound Escherichia coli maltose acetyltransferase in wild-type and amf potato plants. Plant Biotechnol J 5:134–145

    Article  CAS  Google Scholar 

  • Gileadi O (2017) Recombinant protein expression in E. coli: a historical perspective. In: Burgess-Brown NA (ed) Heterologous gene expression in E.coli: methods and protocols. Methods in Molecular Biology, vol 1586. Springer New York, New York, pp 3–10

    Chapter  Google Scholar 

  • Hotze M, Schröder G, Schröder J (1995) Cinnamate 4-hydroxylase from Catharanthus roseus, and a strategy for the functional expression of plant cytochrome P450 proteins as translational fusions with P450 reductase in Escherichia coli. FEBS Lett 374:345–350

    Article  CAS  PubMed  Google Scholar 

  • Iacovelli R, Sokolova N, Haslinger K (2022) Striving for sustainable biosynthesis: discovery, diversification, and production of antimicrobial drugs in Escherichia coli. Biochem Soc Trans 50:1315–1328

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Irmler S, Schröder G, St-Pierre B, Crouch NP, Hotze M, Schmidt J, Strack D, Matern U, Schröder J (2000) Indole alkaloid biosynthesis in Catharanthus roseus: new enzyme activities and identification of cytochrome P450 CYP72A1 as secologanin synthase. Plant J 24:797–804

    Article  CAS  PubMed  Google Scholar 

  • Leggio LL, Degan FD, Poulsen P, Andersen SM, Larsen S (2003) The structure and specificity of Escherichia coli maltose acetyltransferase give new insight into the LacA family of acyltransferases. Biochemistry 42:5225–5235

    Article  PubMed  Google Scholar 

  • Leggio LL, Degan FD, Poulsen P, Sørensen SO, Harlow K, Harris P, Larsen S (2001) Crystallization and preliminary X-ray analysis of maltose O-acetyltransferase. Acta Cryst D57:1915–1918

    Google Scholar 

  • Lewendon A, Ellis J, Shaw WV (1995) Structural and mechanistic studies of galactoside acetyltransferase, the Escherichia coli LacA gene product. J Biol Chem 270:26326–26331

    Article  CAS  PubMed  Google Scholar 

  • Liu M, Kong J (2018) The enzymatic biosynthesis of acylated steroidal glycosides and their cytotoxic activity. Acta Pharm Sin B 8:981–994

    Article  PubMed  PubMed Central  Google Scholar 

  • Lo T, Teo WS, Ling H, Chen B, Kang A, Chang MW (2013) Microbial engineering strategies to improve cell viability for biochemical production. Biotechnol Adv 31:903–914

    Article  CAS  PubMed  Google Scholar 

  • Mori H, Kataoka M, Yang X (2022) Past, present, and future of genome modification in Escherichia coli. Microorganisms 10:1835

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Mosa A, Hutter MC, Zapp J, Bernhardt R, Hannemann F (2015) Regioselective acetylation of C21 hydroxysteroids by the bacterial chloramphenicol acetyltransferase I. ChemBioChem 16:1670–1679

    Article  CAS  PubMed  Google Scholar 

  • Oger C, Marton Z, Brinkmann Y, Bultel-Poncé V, Durand T, Graber M, Galano J (2010) Lipase-catalyzed regioselective monoacetylation of unsymmetrical 1,5-primary diols. J Organomet Chem 75:1892–1897

    Article  CAS  Google Scholar 

  • Park HG, Do JH, Chang HN (2003) Regioselective enzymatic acylation of multi-hydroxyl compounds in organic synthesis. Biotechnol Bioprocess Eng 8:1–8

    Article  CAS  Google Scholar 

  • Pérez JA, Boluda C, López H, Trujillo JM, Hernández JM (2004) Regioselective enzymatic acetylation of the aglycone moiety of a secoiridoid glucoside. Two new secoiridoid glucoside acetates. Chem Pharm Bull 52:1123–1124

    Article  Google Scholar 

  • Pérez JA, Trujillo JM, López H, Aragón Z, Boluda C (2009) Regioselective enzymatic acylation and deacetylation of secoiridoid glucoside. Chem Pharm Bull 57:882–884

    Article  Google Scholar 

  • Qu X (2014) Camptothecin biosynthesis (I) – functional characterization of CaCPR and CaSLS. Dissertation, University of Chinese Academy of Sciences

    Google Scholar 

  • Qu X, Pu X, Chen F, Yang Y, Yang L, Zhang G, Luo Y (2015) Molecular cloning, heterologous expression, and functional characterization of an NADPH-cytochrome P450 reductase gene from Camptotheca acuminata, a camptothecin-producing plant. PLoS One 10:e0135397

    Article  PubMed  PubMed Central  Google Scholar 

  • Roderich SL (2005) The lac operon galactoside acetyltransferase. C R Biologies 328:568–575

    Article  Google Scholar 

  • Sun J, Chen L, Lou B, Bai Y, Yu X, Zhao M, Wang Z (2017) Acetylation and deacetylation for sucralose preparation by a newly isolated Bacillus amyloliquefaciens WZS01. J Biosci Bioeng 123:576–580

    Article  CAS  PubMed  Google Scholar 

  • Terpe K (2006) Overview of bacterial expression systems for heterologous protein production: from molecular and biochemical fundamentals to commercial systems. Appl Microbiol Biotechnol 72:211–222

    Article  CAS  PubMed  Google Scholar 

  • Torres P, Poveda A, Jimenez-Barbero J, Ballesteros A, Plou FJ (2010) Regioselective lipase-catalyzed synthesis of 3-O-acyl derivatives of resveratrol and study of their antioxidant properties. J Agric Food Chem 58:807–813

    Article  CAS  PubMed  Google Scholar 

  • Wang X, Olsen LR, Roderick SL (2002) Structure of the lac operon galactoside acetyltransferase. Structure 10:581–588

    Article  CAS  PubMed  Google Scholar 

  • Wang Z, Sun J, Yang Q, Yang J (2020) Metabolic engineering Escherichia coli for the production of lycopene. Molecules 25:3136

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Wu S, Chen W, Lu S, Zhang H, Yin L (2022) Metabolic engineering of shikimic acid biosynthesis pathway for the production of shikimic acid and its branched products in microorganisms: advances and prospects. Molecules 27:4779

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Yang D, Park SY, Park YS, Eun H, Lee SY (2020) Metabolic engineering of Escherichia coli for natural product biosynthesis. Trends Biotechnol 38:745–765

    Article  CAS  PubMed  Google Scholar 

  • Yang Y, Li W, Pang J, Jiang L, Qu X, Pu X, Zhang G, Luo Y (2019) Bifunctional cytochrome P450 enzymes involved in camptothecin biosynthesis. ACS Chem Biol 14:1091–1096

    Article  CAS  PubMed  Google Scholar 

  • Zabin I, Kepes A, Monod J (1959) On the enzymic acetylation of isopropyl-β-D-thiogalactoside and its association with galactoside-permease. Biochem Biophys Res Commun 1:289–292

    Article  CAS  Google Scholar 

  • Zabin I, Kepes A, Monod J (1962) Thiogalactoside transacetylase. J Biol Chem 237:253–257

    Article  CAS  PubMed  Google Scholar 

  • Zhang X, Xu Q, Xiao H, Liang X (2003) Iridoid glucosides from Strychnos nux-vomica. Phytochemistry 64:1341–1344

    Article  CAS  PubMed  Google Scholar 

Download references

Funding

This work was supported in part by the National Natural Science Foundation of China (22177112), the Natural Science Foundation of Sichuan Province (2022NSFC0616), the Biological Resources Programme, the Chinese Academy of Sciences (KFJ-BRP-008), and the CAS Key Laboratory of Environmental and Applied Microbiology & Environmental Microbiology Key Laboratory of Sichuan Province, Chengdu Institute of Biology, Chinese Academy of Sciences (KLEAMCAS202301).

Author information

Authors and Affiliations

  1. CAS Key Laboratory of Environmental and Applied Microbiology, Environmental Microbiology Key Laboratory of Sichuan Province, Center for Natural Products Research, Chengdu Institute of Biology, Chinese Academy of Sciences, Chengdu, 610041, China

    Yi Li, Xuefei Wang, Zhan Liu, Yun Yang, Liangzhen Jiang, Xixing Qu, Xiang Pu & Yinggang Luo

  2. University of Chinese Academy of Sciences, Beijing, 100049, China

    Yi Li, Xuefei Wang & Zhan Liu

Authors
  1. Yi Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Xuefei Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Zhan Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Yun Yang
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Liangzhen Jiang
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Xixing Qu
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Xiang Pu
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Yinggang Luo
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

YL conceived and designed research. YLi, XW, ZL, YY, LJ, ZL, XQ, and XP conducted the experiments. YLi, YY, XP, and YL analyzed the data. YLi, YY, and YL wrote the manuscript. All authors read and approved the manuscript.

Corresponding author

Correspondence to Yinggang Luo.

Ethics declarations

Conflict of interest

The authors declare no competing interests.

Human and animal rights

This article does not contain any studies with human participants or animals performed by any of the authors.

Additional information

Publisher’s Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Supplementary information

ESM 1

(PDF 1073 kb)

Rights and permissions

Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Li, Y., Wang, X., Liu, Z. et al. Regioselective O-acetylation of various glucosides catalyzed by Escherichia coli maltose O-acetyltransferase. Appl Microbiol Biotechnol 107, 7031–7042 (2023). https://doi.org/10.1007/s00253-023-12790-z

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00253-023-12790-z

Keywords

Search

Navigation