Recent insights in microbial exopolysaccharide biosynthesis and engineering strategies

Current Opinion in Biotechnology

Volumn 53, Issue , 2018, Pages 130-136

  Schmid, Jochen ^a  

^a TECHNICAL UNIVERSITY OF MUNICH   (Germany)

Author keywords

[No Author keywords available]

Indexed keywords

BIOSYNTHESIS; ENZYMES; METABOLITES; POLYSACCHARIDES;

BIOSYNTHESIS PATHWAYS; COMBINATORIAL ASSEMBLY; EXOPOLYSACCHARIDES; GLYCOSYLTRANSFERASES; MICROBIAL POLYSACCHARIDE; OVER-EXPRESSION; REPEATING UNIT; SYNTHASES;

BIOCHEMISTRY;

ABC TRANSPORTER; ALGINIC ACID; BIOPOLYMER; CELLULOSE; DEXTRAN; DIBUTYRYL CYCLIC GMP; EPIMERASE; EXOPOLYSACCHARIDE; GLYCAN; GLYCOSYLTRANSFERASE; LEVAN; MONOMER; UNCLASSIFIED DRUG; WZX PROTEIN; BACTERIAL POLYSACCHARIDE;

BACTERIAL CHROMOSOME; CRYSTAL STRUCTURE; GENE REARRANGEMENT; KLEBSIELLA; MOLECULAR WEIGHT; NONHUMAN; OPEN READING FRAME; OPERON; POLYMER PRODUCTION; POLYMERIZATION; POLYSACCHARIDE STRUCTURE; PRIORITY JOURNAL; PROTEIN ENGINEERING; REVIEW; STOICHIOMETRY; BACTERIAL GENOME; BIOLOGICAL MODEL; BIOSYNTHESIS; GENETICS; METABOLIC ENGINEERING; PROCEDURES;

BIOSYNTHETIC PATHWAYS; GENE REARRANGEMENT; GENOME, BACTERIAL; METABOLIC ENGINEERING; MODELS, BIOLOGICAL; POLYSACCHARIDES, BACTERIAL;

EID: 85041463113     PISSN: 09581669     EISSN: 18790429     Source Type: Journal    
DOI: 10.1016/j.copbio.2018.01.005     Document Type: Review

References (55)

1. Corbett, D., Roberts, I.S., The role of microbial polysaccharides in host–pathogen interaction. F1000 Biol Rep, 1, 2009, 30.
2. Limoli, D.H., Jones, C.J., Wozniak, D.J., Bacterial extracellular polysaccharides in biofilm formation and function. Microbiol Spec, 3, 2015, 10.1128/microbiolspec.MB-0011-2014.
3. Buttimer, C., McAuliffe, O., Ross, R.P., Hill, C., O'Mahony, J., Coffey, A., Bacteriophages and bacterial plant diseases. Front Microbiol, 8, 2017, 34 This article summarizes the different plant pathogen mechanisms and displays the function of polysaccharides in plant pathogenicity.
4. Cress, B.F., Englaender, J.A., He, W., Kasper, D., Linhardt, R.J., Koffas, M.A., Masquerading microbial pathogens: capsular polysaccharides mimic host-tissue molecules. FEMS Microbiol Rev 38 (2014), 660–697.
5. Freitas, F., Alves, V.D., Reis, M.A.M., Bacterial polysaccharides: production and applications in cosmetic industry. Ramawat, K.G., Mérillon, J.-M., (eds.) Polysaccharides: Bioactivity and Biotechnology, 2014, Springer International Publishing, Cham, 1–24.
6. Lapasin, R., Pricl, S., Industrial applications of polysaccharides. Lapasin, R., Pricl, S., (eds.) Rheology of Industrial Polysaccharides: Theory and Applications, 1995, Springer, US, 134–161.
7. Paul, F., Morin, A., Monsan, P., Microbial polysaccharides with actual potential industrial applications. Biotechnol Adv 4 (1986), 245–259.
8. Lourenco, S.C., Torres, C.A.V., Nunes, D., Duarte, P., Freitas, F., Reis, M.A.M., Fortunato, E., Moldao-Martins, M., da Costa, L.B., Alves, V.D., Using a bacterial fucose-rich polysaccharide as encapsulation material of bioactive compounds. Int J Biol Macromol 104 (2017), 1099–1106.
9. Freitas, F., Torres, C.A.V., Reis, M.A.M., Engineering aspects of microbial exopolysaccharide production. Bioresour Technol 245:Part B (2017), 1674–1683 This review summarizes several aspects of polysaccharide engineering, including influence of media compounds, process parameters as well as down-stream processing. The use of renewable resources as alternative fermentation substrate is also discussed. This manuscript is recommended to obtain an updated overview on microbial EPS production.
10. Rühmann, B., Schmid, J., Sieber, V., High throughput exopolysaccharide screening platform: from strain cultivation to monosaccharide composition and carbohydrate fingerprinting in one day. Carbohydr Polym 122 (2015), 212–220.
11. Baker, P., Whitfield, G.B., Hill, P.J., Little, D.J., Pestrak, M.J., Robinson, H., Wozniak, D.J., Howell, P.L., Characterization of the Pseudomonas aeruginosa glycoside hydrolase PslG reveals that its levels are critical for Psl polysaccharide biosynthesis and biofilm formation. J Biol Chem 290 (2015), 28374–28387.
12. Schmid, J., Sieber, V., Enzymatic transformations involved in the biosynthesis of microbial exo-polysaccharides based on the assembly of repeat units. Chembiochem 16 (2015), 1141–1147.
13. Islam, S.T., Lam, J.S., Synthesis of bacterial polysaccharides via the Wzx/Wzy-dependent pathway. Can J Microbiol 60 (2014), 697–716.
14. Hong, Y., Liu, M.A., Reeves, P.R., Progress in our understanding of Wzx flippase for translocation of bacterial membrane lipid-linked oligosaccharide. J Bacteriol, 2017, 10.1128/JB.00154-17 This current summary of latest insights on the Wzx protein family describes and compares the different Wzx proteins and especially the mechanisms as well as specificities. Based on these conclusions it is obvious that highly specific as well as promiscuous Wzx variants exist.
15. Liu, M.A., Stent, T.L., Hong, Y., Reeves, P.R., Inefficient translocation of a truncated O unit by a Salmonella Wzx affects both O-antigen production and cell growth. FEMS Microbiol Lett, 2015, 362.
16. Whitfield, C., Polymerases: glycan chain-length control. Nat Chem Biol 6 (2010), 403–404.
17. Rehm, B.H.A., Bacterial polymers: biosynthesis, modifications and applications. Nat Rev Micro 8 (2010), 578–592.
18. Cuthbertson, L., Mainprize, I.L., Naismith, J.H., Whitfield, C., Pivotal roles of the outer membrane polysaccharide export and polysaccharide copolymerase protein families in export of extracellular polysaccharides in gram-negative bacteria. Microbiol Mol Biol Rev 73 (2009), 155–177.
19. Schmid, J., Sperl, N., Sieber, V., A comparison of genes involved in sphingan biosynthesis brought up to date. Appl Microbiol Biotechnol 98 (2014), 7719–7733.
20. Schmid, J., Sieber, V., Rehm, B., Bacterial exopolysaccharides: biosynthesis pathways and engineering strategies. Front Microbiol, 6, 2015, 496 This review summarizes the insights into bacterial EPS production and biosynthesis up to 2015 in a very comprehensive way and suggest first engineering strategies for tailored EPS production. Based on the most prominent representatives, the different pathways are described in detail, and their EPS encoding operons and the resulting chemical structures are compared.
21. Kos, V., Cuthbertson, L., Whitfield, C., The Klebsiella pneumoniae O2a antigen defines a second mechanism for O antigen ATP-binding cassette transporters. J Biol Chem 284 (2009), 2947–2956.
22. Mann, E., Mallette, E., Clarke, B.R., Kimber, M.S., Whitfield, C., The Klebsiella pneumoniae O12 ATP-binding cassette (ABC) transporter recognizes the terminal residue of its O-antigen polysaccharide substrate. J Biol Chem 291 (2016), 9748–9761.
23. Hagelueken, G., Huang, H., Clarke, B.R., Lebl, T., Whitfield, C., Naismith, J.H., Structure of WbdD: a bifunctional kinase and methyltransferase that regulates the chain length of the O antigen in Escherichia coli O9a. Mol Microbiol 86 (2012), 730–742.
24. King, J.D., Berry, S., Clarke, B.R., Morris, R.J., Whitfield, C., Lipopolysaccharide O antigen size distribution is determined by a chain extension complex of variable stoichiometry in Escherichia coli O9a. Proc Natl Acad Sci USA 111 (2014), 6407–6412.
25. Williams, D.M., Ovchinnikova, O.G., Koizumi, A., Mainprize, I.L., Kimber, M.S., Lowary, T.L., Whitfield, C., Single polysaccharide assembly protein that integrates polymerization, termination, and chain-length quality control. Proc Natl Acad Sci USA 114 (2017), E1215–E1223 This research article characterises the different activities of the WbbB protein from Raoultella terrigena for the first time and compare it with other homologues. This remarkable protein family contains multiple glycosyltransferase modules for glycan polymerization and chain termination. These glycosyltransferase modules often represent novel glycosyltransferase families. The authors were able to assign the chain length regulation to a molecular marker, which separates polymerase and terminator activity and thus might enable novel strategies for glycoengineering.
26. Perez, C., Gerber, S., Boilevin, J., Bucher, M., Darbre, T., Aebi, M., Reymond, J.L., Locher, K.P., Structure and mechanism of an active lipid-linked oligosaccharide flippase. Nature 524 (2015), 433–438 This article give impressive insights on flippase structure and mechanism based on experimental results. It is shown that the mechanism of the ABC transporter PglK is distinct from the classical alternating-access model as suggested for other transporters, and is mainly based on inward-facing and outward-facing conformations, whereas only the outward-facing conformation is necessary for flipping of the repeat unit.
27. Kong, L., Almond, A., Bayley, H., Davis, B.G., Chemical polyglycosylation and nanolitre detection enables single-molecule recapitulation of bacterial sugar export. Nat Chem 8 (2016), 461–469 This article describes the mechanism of sugar export by using a synthetic nanolitre detection system that makes use of the bilayer interface between two droplets, thus mimicking the natural process. On the basis of this approach it was shown that the Wza protein of E. coli translocates tetrasaccharide units but not longer fragments.
28. Locher, K.P., Mechanistic diversity in ATP-binding cassette (ABC) transporters. Nat Struct Mol Biol 23 (2016), 487–493.
29. Woodward, L., Naismith, J.H., Bacterial polysaccharide synthesis and export. Curr Opin Struct Biol 40 (2016), 81–88.
30. Augimeri, R.V., Varley, A.J., Strap, J.L., Establishing a role for bacterial cellulose in environmental interactions: lessons learned from diverse biofilm-producing proteobacteria. Front Microbiol, 6, 2015, 1282.
31. McNamara, J.T., Morgan, J.L.W., Zimmer, J., A molecular description of cellulose biosynthesis. Annu Rev Biochem 84 (2015), 895–921 This review article summarizes the impressive insights in the structure and the mechanism of the cellulase synthase complex by mainly own results of the authors. Especially the highly specific inverting mechanism of the GT domain is depicted together with the whole structure of the complex.
32. Morgan, J.L., Strumillo, J., Zimmer, J., Crystallographic snapshot of cellulose synthesis and membrane translocation. Nature 493 (2013), 181–186.
33. Morgan, J.L., McNamara, J.T., Zimmer, J., Mechanism of activation of bacterial cellulose synthase by cyclic di-GMP. Nat Struct Mol Biol 21 (2014), 489–496.
34. Jang, W.D., Hwang, J.H., Kim, H.U., Ryu, J.Y., Lee, S.Y., Bacterial cellulose as an example product for sustainable production and consumption. Microb Biotechnol 10:5 (2017), 1181–1185.
35. Hay, I.D., Rehman, Z.U., Moradali, M.F., Wang, Y., Rehm, B.H.A., Microbial alginate production, modification and its applications. Microb Biotechnol 6 (2013), 637–650.
36. Ertesvåg, H., Alginate-modifying enzymes: biological roles and biotechnological uses. Front Microbiol, 6, 2015.
37. Wang, Y., Moradali, M.F., Goudarztalejerdi, A., Sims, I.M., Rehm, B.H.A., Biological function of a polysaccharide degrading enzyme in the periplasm. Sci Rep, 6, 2016, 31249.
38. Maleki, S., Almaas, E., Zotchev, S., Valla, S., Ertesvåg, H., Alginate biosynthesis factories in Pseudomonas fluorescens: localization and correlation with alginate production level. Appl Environ Microb 82 (2016), 1227–1236 This article demonstrates for the first time the localization and spatial distribution of the alginate biosynthesis machinery in Pseudomonas aeruginosa by immunogold labeling of the AlgE-FLAG tagged protein. The correlation of alginate production and number of alginate biosynthesis complexes is impressively shown.
39. Ua-Arak, T., Jakob, F., Vogel, R.F., Fermentation pH modulates the size distributions and functional properties of Gluconobacter albidus TMW 2.1191 levan. Front Microbiol, 8, 2017, 807.
40. Leemhuis, H., Pijning, T., Dobruchowska, J.M., van Leeuwen, S.S., Kralj, S., Dijkstra, B.W., Dijkhuizen, L., Glucansucrases: three-dimensional structures, reactions, mechanism, alpha-glucan analysis and their implications in biotechnology and food applications. J Biotechnol 163 (2013), 250–272.
41. Srikanth, R., Reddy, C.H., Siddartha, G., Ramaiah, M.J., Uppuluri, K.B., Review on production, characterization and applications of microbial levan. Carbohydr Polym 120 (2015), 102–114.
42. Hu, Y., Winter, V., Chen, X.Y., Ganzle, M.G., Effect of acceptor carbohydrates on oligosaccharide and polysaccharide synthesis by dextransucrase DsrM from Weissella cibaria. Food Res Int 99 (2017), 603–611.
43. Pan, Y.J., Lin, T.L., Chen, C.T., Chen, Y.Y., Hsieh, P.F., Hsu, C.R., Wu, M.C., Wang, J.T., Genetic analysis of capsular polysaccharide synthesis gene clusters in 79 capsular types of Klebsiella spp. Sci Rep, 5, 2015, 15573.
44. Zeidan, A.A., Poulsen, V.K., Janzen, T., Buldo, P., Derkx, P.M.F., Oregaard, G., Neves, A.R., Polysaccharide production by lactic acid bacteria: from genes to industrial applications. FEMS Microbiol Rev 41 (2017), S168–S200.
45. Cheng, X., Zheng, X., Zhou, X., Zeng, J., Ren, Z., Xu, X., Cheng, L., Li, M., Li, J., Li, Y., Regulation of oxidative response and extracellular polysaccharide synthesis by a diadenylate cyclase in Streptococcus mutans. Environ Microbiol 18 (2016), 904–922.
46. Rutering, M., Schmid, J., Ruhmann, B., Schilling, M., Sieber, V., Controlled production of polysaccharides-exploiting nutrient supply for levan and heteropolysaccharide formation in Paenibacillus sp. Carbohydr Polym 148 (2016), 326–334.
47. Pérez-Mendoza, D., Rodríguez-Carvajal, M.Á., Romero-Jiménez, L., Farias GdA, Lloret, J., Gallegos, M.T., Sanjuán, J., Novel mixed-linkage β-glucan activated by c-di-GMP in Sinorhizobium meliloti. Proc Natl Acad Sci USA 112 (2015), E757–E765 A new EPS variant is produced by overexpression of c-di-GMP in S. meliloti as described in that article. Next to the proof of the inducing effects of c-di-GMP on the EPS biosynthesis of S. meliloti, a very rare and highly interesting EPS structure is described.
48. Liang, Z.-X., The expanding roles of c-di-GMP in the biosynthesis of exopolysaccharides and secondary metabolites. Nat Prod Rep 32 (2015), 663–683.
49. Pérez-Mendoza, D., Bertinetti, D., Lorenz, R., Gallegos, M.-T., Herberg, F.W., Sanjuán, J., A novel c-di-GMP binding domain in glycosyltransferase BgsA is responsible for the synthesis of a mixed-linkage β-glucan. Sci Rep, 7, 2017, 8997 In this study the binding of c-di-GMP by the cytoplasmic C-terminal domain of BgsA of S. meliloti is shown, which does not contain a PILZ motif for c-di-GMP recognition. By mutagenesis of the BgsA protein its role in c-di-GMP binding as well as its GT activity is shown.
50. Schäper, S., Steinchen, W., Krol, E., Altegoer, F., Skotnicka, D., Søgaard-Andersen, L., Bange, G., Becker, A., AraC-like transcriptional activator CuxR binds c-di-GMP by a PilZ-like mechanism to regulate extracellular polysaccharide production. Proc Natl Acad Sci USA 114 (2017), E4822–E4831 Another novel c-di-GMP-responsive protein is described in S. meliloti belonging to the AraC like transcription factor CuxR. C-di-GMP binding is realized by a similar mechanism as in the PILZ domain, but is based on interplay of a Cupin domain and the conserved RxxxR motif, and therefore lacks sequence similarity. This study displays the molecular evolution of c-di-GMP binding proteins.
51. Schmid, J., Heider, D., Wendel, N.J., Sperl, N., Sieber, V., Bacterial glycosyltransferases: challenges and opportunities of a highly diverse enzyme class toward tailoring natural products. Front Microbiol, 7, 2016, 182.
52. Becker, A., Challenges and perspectives in combinatorial assembly of novel exopolysaccharide biosynthesis pathways. Front Microbiol, 6, 2015, 687 This review article summarizes the challenges of designing novel synthetic EPS biosynthesis pathways by limitations of GT predictability as well as the specificity of the Wzx and Wzy proteins, but at the same time describes the high potential of synthetic biology approaches.
53. Florea, M., Hagemann, H., Santosa, G., Abbott, J., Micklem, C.N., Spencer-Milnes, X., de Arroyo Garcia, L., Paschou, D., Lazenbatt, C., Kong, D., et al. Engineering control of bacterial cellulose production using a genetic toolkit and a new cellulose-producing strain. Proc Natl Acad Sci USA 113 (2016), E3431–E3440 This article is one of the first reports on using synthetic biology approaches to regulate cellulose production and is the result of the iGEM competition, thus proofing the principle of tailoring EPS-biosynthesis. Additionally, the targeted in vivo functionalisation of cellulose by binding different proteins is realised, showing the potential for realizing novel product properties.
54. Rütering, M., Cress, B.F., Schilling, M., Rühmann, B., Koffas, M.A.G., Sieber, V., Schmid, J., Tailor-made exopolysaccharides — CRISPR-Cas9 mediated genome editing in Paenibacillus polymyxa. Synth Biol, 2(1), 2017, ysx007 One of the first reports on using CRISPR-Cas9 for the engineering of polysaccharide structures in a novel described biosyntesis pathway. Additionally, this manuscript describes an EPS-biosynthesis encoding operon of extreme length which carries some genes in duplicate (e.g. wzx, ugdh) and a large amount of glycosyltransferases, indicating the possibility to produce different exopolysaccharides or repeatig units by use of merged operons.
55. Schatschneider, S., Schneider, J., Blom, J., Létisse, F., Niehaus, K., Goesmann, A., Vorhölter, F.-J., Systems and synthetic biology perspective of the versatile plant–pathogenic and polysaccharide-producing bacterium Xanthomonas campestris. Microbiology 163 (2017), 1117–1144 This review article summarizes the research on xanthan biosynthesis and links it to the different omics technologies and bioinformatic tools which get available throughout the last years. In brief a complete analysis pipeline for modelling the biosynthesis pathway is given and is highly recommended to the readers interested in systems and synthetic biology.