Large-scale production of N-acetyllactosamine through bacterial coupling

Endo, Tetsuo; Koizumi, Satoshi; Tabata, Kazuhiko; Kakita, Shingo; Ozaki, Akio

doi:10.1016/s0008-6215(99)00050-6

Cited by 63 publications

(26 citation statements)

References 15 publications

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“…In such experiments, several recombinant bacterial strains expressing enzymes involved in the catalytic pathway of particular carbohydrates were coupled. The development of a metabolically engineered strain of Corynebacterium ammoniagenes with highlevel UTP production and UDP-galactose (UDP-Gal) regeneration was a key contribution to the present work (11)(12)(13)20). Subsequently, an alternate pathway for UDP-Gal formation from sucrose and UDP has been developed.…”

mentioning

confidence: 99%

P1 Trisaccharide (Galα1,4Galβ1,4GlcNAc) Synthesis by Enzyme Glycosylation Reactions Using Recombinant Escherichia coli

Liu

Zhang

et al. 2003

Appl Environ Microbiol

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The frequency of Escherichia coli infection has lead to concerns over pathogenic bacteria in our food supply and a demand for therapeutics. Glycolipids on gut cells serve as receptors for the Shiga-like toxin produced by E. coli. Oligosaccharide moiety analogues of these glycolipids can compete with receptors for the toxin, thus acting as antibacterials. An enzymatic synthesis of the P1 trisaccharide (Gal␣1,4Gal␤1,4GlcNAc), one of the oligosaccharide analogues, was assessed in this study. In the proposed synthetic pathway, UDP-glucose was generated from sucrose with an Anabaena sp. sucrose synthase and then converted with an E. coli UDP-glucose 4-epimerase to UDP-galactose. Two molecules of galactose were linked to N-acetylglucosamine subsequently with a Helicobacter pylori ␤-l,4-galactosyltransferase and a Neisseria meningitidis ␣-1,4-galactosyltransferase to produce one molecule of P1 trisaccharide. The four enzymes were coexpressed in a single genetically engineered E. coli strain that was then permeabilized and used to catalyze the enzymatic reaction. P1 trisaccharide was accumulated up to 50 mM (5.4 g in a 200-ml reaction volume), with a 67% yield based on the consumption of N-acetylglucosamine. This study provides an efficient approach for the preparative-scale synthesis of P1 trisaccharide with recombinant bacteria.Recently, vaccines have been introduced to prevent illness from pathogenic Escherichia coli, yet there remains a need for therapeutics to treat acute infection (27). Unlike endogenous E. coli from our intestine, the pathogenic strain (O157:H7) has acquired a gene from Shigella which produces a toxic protein. This Shiga-like toxin binds to glycolipid receptors on cells of the gut wall and can ultimately lead to dysentery, hemorrhagic colitis, and sometimes life-threatening complications (32). The attachment of the microbial protein onto mammalian cells through surface carbohydrates initiates a successful infection (18). The Shiga-like toxin will also bind other carbohydrate derivatives, including globotriosylceramide (Gal␣1,4Gal␤1, 4Glc-ceramide) found on the cell surface (4,(24)(25)(26)44) and trisaccharide analogues, such as globotriose (Gal␣1,4Gal␤1, 4Glc) and the trisaccharide moiety of P1 antigen (Gal␣1, 4Gal␤1,4GlcNAc) (1). The mechanism of action for the proposed therapy involves preventing the toxin from binding to cell receptors by introducing these molecules to the site of infection. Instead of binding to cell receptors, the toxic peptides bind to the carbohydrate analogues and are removed from the body without causing harm. Of the three carbohydrate moieties mentioned above, P1 trisaccharide is the most effective therapeutic. Although synthesis remains a limiting factor, if available in large quantities these trisaccharides have considerable potential as antiadhesive agents in the treatment of pathogenic E. coli infections (17,36,37).Chemical syntheses of oligosaccharides have always been complicated by their structural complexity and have traditionally involved laborious protection p...

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confidence: 99%

P1 Trisaccharide (Galα1,4Galβ1,4GlcNAc) Synthesis by Enzyme Glycosylation Reactions Using Recombinant Escherichia coli

Liu

Zhang

et al. 2003

Appl Environ Microbiol

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“…Metabolically engineered strains of Corynebacterium ammoniagenes were shown to be useful for the overproduction of deoxycytidine (Lee et al 2011), uridine 5 0 -monophosphate (Wang et al 2007), and 5 0 -inosine monophosphate (Abbouni et al 2004). A large-scale production system of N-acetyllactosamine, a core structure of various oligosaccharides, was established by a whole-cell reaction through the combination of recombinant Escherichia coli strains and Corynebacterium ammoniagenes (Endo et al 1999). Likewise, UDP-galactose and globotriose can be produced on a large scale by coupling Corynebacterium ammoniagenes and metabolically engineered Escherichia coli strains (Koizumi et al 1998).…”

Section: Applicationmentioning

confidence: 99%

The Family Corynebacteriaceae

Tauch

Sandbote

2014

The Prokaryotes

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“…This technology was developed by Kyowa Hakko Kogyo Co. Ltd. in Japan [31][32][33][34][35]. The key in Kyowa Hakko's technology for the large-scale production of oligosaccharides is a C. ammoniagenes bacterial strain engineered to efficiently convert inexpensive orotic acid to UTP (Scheme 3).…”

Section: Scheme 2 Biosynthesis Of Oligosaccharide By Glycosyltransfementioning

confidence: 99%

Approaches to the Enzymatic Carbohydrate Synthesis

Kowal

Shao

Mei

et al. 2004

ACS Symposium Series

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Polysaccharides are the least appreciated class of macromolecules though they constitute the largest part of the planet's bio-mass. This large bulk has perpetuated a compelling reason for their neglect: carbohydrates were thought to be biochemically uninteresting serving as structural and energy storage molecules. The second more insidious and perhaps contradictory reason for the relative slowness in carbohydrate research is that biochemically important carbohydrates are difficult to study. They seem haphazardly put together, without a template, creating enormous complexity that, at first, did not suggest specific functions. The past three decades have seen enormous strides in the understanding of the importance of carbohydrates feeding the quickly expanding and vibrant field of enzymatic carbohydrate synthesis. The clear goal of the research in this field is to supplant Nature as the supreme oligosaccharide maker to generate desired saccharides, be it natural or unnatural, in high yields, large quantities and at low cost.

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Large-scale production of N-acetyllactosamine through bacterial coupling

Cited by 63 publications

References 15 publications

P1 Trisaccharide (Galα1,4Galβ1,4GlcNAc) Synthesis by Enzyme Glycosylation Reactions Using Recombinant Escherichia coli

P1 Trisaccharide (Galα1,4Galβ1,4GlcNAc) Synthesis by Enzyme Glycosylation Reactions Using Recombinant Escherichia coli

The Family Corynebacteriaceae

Approaches to the Enzymatic Carbohydrate Synthesis

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