Capsular Polysaccharide Biosynthesis from Recombinant E. coli and Chondroitin Sülfate Production

Ayse Sebnem Erenler

doi:10.14715/cmb/2019.65.6.4

Issue

Vol. 65 No. 6: Issue 6

Issue Published : August 5, 2019

The undersigned hereby assign all rights, included but not limited to copyright, for this manuscript to CMB Association upon its submission for consideration to publication on Cellular and Molecular Biology. The rights assigned include, but are not limited to, the sole and exclusive rights to license, sell, subsequently assign, derive, distribute, display and reproduce this manuscript, in whole or in part, in any format, electronic or otherwise, including those in existence at the time this agreement was signed. The authors hereby warrant that they have not granted or assigned, and shall not grant or assign, the aforementioned rights to any other person, firm, organization, or other entity. All rights are automatically restored to authors if this manuscript is not accepted for publication.

Capsular Polysaccharide Biosynthesis from Recombinant E. coli and Chondroitin Sülfate Production

https://doi.org/10.14715/cmb/2019.65.6.4

Ayse Sebnem Erenler

Department of Biology, Inonu University, 44280 Malatya, Turkey

Corresponding Author(s) : Ayse Sebnem Erenler

sebnem.erenler@inonu.edu.tr

Cellular and Molecular Biology, Vol. 65 No. 6: Issue 6
Article Published : July 31, 2019

Abstract

Chondroitin sulfate (CS) is an important biomedical product. CS is the basic structural component of the mammalian extracellular matrix and is widely used in many applications in the fields of medicine, veterinary medicine, pharmaceuticals and cosmetics. For CS production, mainly animal sources are used. However, in today's conditions, due to various risks and artificial synthesis, there has been an increase in alternative sources of production methods for CS, instead of using animal resources. In this study as a powerful alternative microbial production of CS has been targeted. By using recombinant E. coli strains to integrate VHb /vgb+ and kfo+ systems, the aim was to obtain high purity CS from reliable biotechnological processes. Plasmid pUC8:15 bearing the vgb gene region, and plasmid pETM6-PACF carrying the kfoA, kfoC and kfoF genes responsible for chondroitin synthesis, were transferred to E. coli bacteria. Microbial CS was obtained by adding sulfate groups to chondroitin acquired after the treatments. The results were confirmed by HPLC and NMR analyses. The product, compared to its counterparts, was found to be an effective drug, potentially with a low molecular weight value.

Keywords

Chondroitin Nonfructosylated chondroitin Chondroitin sulfate Vitreoscilla hemoglobin vgb gene.

Erenler, A. S. (2019). Capsular Polysaccharide Biosynthesis from Recombinant E. coli and Chondroitin Sülfate Production. Cellular and Molecular Biology, 65(6), 17–21. https://doi.org/10.14715/cmb/2019.65.6.4

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References

References
Schiraldi, C, Cimini D, De Rosa M. Production of chondroitin sulfate and chondroitin. Applied Microbiology and Biotechnology 2010; 87: 1209–1220.
Cress B F, Englaender JA, He W, Kasper D, Linhardt R J, Koffas M. Masquerading microbial pathogens: Capsular polysaccharides mimic host-tissue molecules. FEMS Microbiology 2014; Reviews 38: 660–697.
Martens EC, Koropatkin NM, Smith TJ, Gordon JI. Complex glycan catabolism by the human gut microbiota: the Bacteroidetes Sus-like paradigm. J Biol Chem 2009; 284 (37): 24673–24677.
He W, Fu L, Li G, Andrew Jones J, Linhardt RJ, Koffas M. Production of chondroitin in metabolically engineered E. coli. Metab Eng 2015; 27: 92-100.
Erenler S, Gencer S, Geçkil H, Stark B, Webster D. Cloning and expression of the Vitreoscilla hemoglobin gene in Enterobacter aerogenes: Effect on cell growth and oxygen uptake. Applied Biochemistry and Microbiology 2004; 40: 241-248.
Kaur R, Ahuja S, Anand A, Singh B, Dikshit KL. Functional implications of the proximal site hydrogen bonding network in Vitreoscilla hemoglobin (VHb): Role of Tyr95 (G5) and Tyr126 (H12). FEBS Letters 2008; 582: 3494-3500.
Geçkil H, Stark B. Webster D. Cell growth and oxygen uptake of Escherichia coli and Pseudomonas aeruginosa are differently effected by the genetically engineered Vitreoscilla hemoglobin gene. Journal of Biotechnology 2001; 85: 57-66.
Chi PY, Webster D A, Stark BC. Vitreoscilla hemoglobin aids respiration under hypoxic conditions in its native host. Microbiological Research 2009; 164: 267-275. 5.
Rajendra P, Dikshit KL, Yixiang L, Webster DA. The bacterial hemoglobin from Vitreoscilla can support the aerobic growth of Escherichia coli lacking terminal oxidases. Archives of Biochemistry and Biophysics 1992; 293: 241-245.
Dikshit KL, Webster DA, Cloning Characterization and expression of the bacterial globin gene from Vitreoscilla in Escherichia coli. Gene 1988;70: 377–386.
Manıatıs T, Frıtsch EF, Sambrook J. Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory Press, 1989.
Fu L, Li G, Yang B, Onishi A, Li L, Sun P, Zhang F, Linhardt RJ. Structural characterization of pharmaceutical heparins prepared from different animal tissues. J. Pharm. Sci 2013; 102: 1447–1457.
Yang B, Chang Y, Weyers AM, Sterner E, Linhardt RJ. Disaccharide analysis of glycosaminoglycan mixtures by ultra-high-performance liquid chromatography–mass spectrometry. J.Chromatogr.A 2012; 122: 591–98.
Gustavo RC.S, Adriana AP, Bianca FG, Ana MFT, Mariana S P, Eduardo V, Paulo ASM. Systematic Analysis of Pharmaceutical Preparations of Chondroitin Sulfate Combined with Glucosamine. Pharmaceuticals 2017; 10(2): 38.
Jıng P, Yi Q, Xiaodong Z, Pazandark A, Frazier SB, Weiser P, Hog L, Lijuan Z. Oversulfated chondroitin sulfate is not the sole contaminant in heparin. Nature Biotechnology 2010; 28(3): 203-207.
Jolly J, Klimaszewski K, Nakanishi Y, Matsubara H, Takahashi T, Nishio K. Microbial-derived chondroitin sulphate. Official Gazette of the United States Patent and Trademark Office Patents, Patent Publication Number: US 2010/0063001 A1. 2010
Campo GM, Avenoso A, Campo S, D'Ascola A, Traina P, Sama D, et al.. Glycosaminoglycans modulate inflammation and apoptosis in LPS-treated chondrocytes. J Cell Biochem 2009a ;106: 83–92.
Campo GM, Avenoso A, Campo S, D'Ascola A, Traina P, Calatroni A. Chondroitin-4-sulphate inhibits NF-kB translocation and caspase activation in collagen-induced arthritis in mice. Osteoarthritis Cartilage 2008; 16: 1474–1483.
Campo GM, Avenoso A, Campo S, Ferlazzo AM, Calatroni AAntioxidant activity of chondroitin sulfate. Adv Pharmacol 2006a; 53: 417-431.
Cress BF, Englaender JA , He W, Kasper D, Linhardt RJ, Koffas MA. Masquerading microbial pathogens: Capsular polysaccharides mimic host-tissue molecules. FEMS Microbiol Rev 2014; 1–38.
Vázquez JA, Rodríguez AI, Montemayor MI, Fraguas J, González MP and Murado MA. Chondroitin Sulfate, Hyaluronic Acid and Chitin/Chitosan Production Using Marine Waste Sources: Characteristics, Applications and Eco-Friendly Processes: A Review Mar. Drugs 2013; 11(3), 747-774.

References

Schiraldi, C, Cimini D, De Rosa M. Production of chondroitin sulfate and chondroitin. Applied Microbiology and Biotechnology 2010; 87: 1209–1220.

Cress B F, Englaender JA, He W, Kasper D, Linhardt R J, Koffas M. Masquerading microbial pathogens: Capsular polysaccharides mimic host-tissue molecules. FEMS Microbiology 2014; Reviews 38: 660–697.

Martens EC, Koropatkin NM, Smith TJ, Gordon JI. Complex glycan catabolism by the human gut microbiota: the Bacteroidetes Sus-like paradigm. J Biol Chem 2009; 284 (37): 24673–24677.

He W, Fu L, Li G, Andrew Jones J, Linhardt RJ, Koffas M. Production of chondroitin in metabolically engineered E. coli. Metab Eng 2015; 27: 92-100.

Erenler S, Gencer S, Geçkil H, Stark B, Webster D. Cloning and expression of the Vitreoscilla hemoglobin gene in Enterobacter aerogenes: Effect on cell growth and oxygen uptake. Applied Biochemistry and Microbiology 2004; 40: 241-248.

Kaur R, Ahuja S, Anand A, Singh B, Dikshit KL. Functional implications of the proximal site hydrogen bonding network in Vitreoscilla hemoglobin (VHb): Role of Tyr95 (G5) and Tyr126 (H12). FEBS Letters 2008; 582: 3494-3500.

Geçkil H, Stark B. Webster D. Cell growth and oxygen uptake of Escherichia coli and Pseudomonas aeruginosa are differently effected by the genetically engineered Vitreoscilla hemoglobin gene. Journal of Biotechnology 2001; 85: 57-66.

Chi PY, Webster D A, Stark BC. Vitreoscilla hemoglobin aids respiration under hypoxic conditions in its native host. Microbiological Research 2009; 164: 267-275. 5.

Rajendra P, Dikshit KL, Yixiang L, Webster DA. The bacterial hemoglobin from Vitreoscilla can support the aerobic growth of Escherichia coli lacking terminal oxidases. Archives of Biochemistry and Biophysics 1992; 293: 241-245.

Dikshit KL, Webster DA, Cloning Characterization and expression of the bacterial globin gene from Vitreoscilla in Escherichia coli. Gene 1988;70: 377–386.

Manıatıs T, Frıtsch EF, Sambrook J. Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory Press, 1989.

Fu L, Li G, Yang B, Onishi A, Li L, Sun P, Zhang F, Linhardt RJ. Structural characterization of pharmaceutical heparins prepared from different animal tissues. J. Pharm. Sci 2013; 102: 1447–1457.

Yang B, Chang Y, Weyers AM, Sterner E, Linhardt RJ. Disaccharide analysis of glycosaminoglycan mixtures by ultra-high-performance liquid chromatography–mass spectrometry. J.Chromatogr.A 2012; 122: 591–98.

Gustavo RC.S, Adriana AP, Bianca FG, Ana MFT, Mariana S P, Eduardo V, Paulo ASM. Systematic Analysis of Pharmaceutical Preparations of Chondroitin Sulfate Combined with Glucosamine. Pharmaceuticals 2017; 10(2): 38.

Jıng P, Yi Q, Xiaodong Z, Pazandark A, Frazier SB, Weiser P, Hog L, Lijuan Z. Oversulfated chondroitin sulfate is not the sole contaminant in heparin. Nature Biotechnology 2010; 28(3): 203-207.

Jolly J, Klimaszewski K, Nakanishi Y, Matsubara H, Takahashi T, Nishio K. Microbial-derived chondroitin sulphate. Official Gazette of the United States Patent and Trademark Office Patents, Patent Publication Number: US 2010/0063001 A1. 2010

Campo GM, Avenoso A, Campo S, D'Ascola A, Traina P, Sama D, et al.. Glycosaminoglycans modulate inflammation and apoptosis in LPS-treated chondrocytes. J Cell Biochem 2009a ;106: 83–92.

Campo GM, Avenoso A, Campo S, D'Ascola A, Traina P, Calatroni A. Chondroitin-4-sulphate inhibits NF-kB translocation and caspase activation in collagen-induced arthritis in mice. Osteoarthritis Cartilage 2008; 16: 1474–1483.

Campo GM, Avenoso A, Campo S, Ferlazzo AM, Calatroni AAntioxidant activity of chondroitin sulfate. Adv Pharmacol 2006a; 53: 417-431.

Cress BF, Englaender JA , He W, Kasper D, Linhardt RJ, Koffas MA. Masquerading microbial pathogens: Capsular polysaccharides mimic host-tissue molecules. FEMS Microbiol Rev 2014; 1–38.

Vázquez JA, Rodríguez AI, Montemayor MI, Fraguas J, González MP and Murado MA. Chondroitin Sulfate, Hyaluronic Acid and Chitin/Chitosan Production Using Marine Waste Sources: Characteristics, Applications and Eco-Friendly Processes: A Review Mar. Drugs 2013; 11(3), 747-774.

Author biographies is not available.