Biotechnological advances in UDP-sugar based glycosylation of small molecules

Bruyn, Frederik De; Maertens, Jo; Beauprez, Joeri; Soetaert, Wim; Mey, Marjan De

doi:10.1016/j.biotechadv.2015.02.005

Cited by 136 publications

(124 citation statements)

References 213 publications

Supporting

Mentioning

120

Contrasting

Order By: Relevance

“…Gene abbreviations: sps=sucrose-phosphate synthase, spp=sucrose-phosphate phosphatase, susy=sucrose synthase, sp=sucrose phosphorylase, frk=fructokinase, pfkb=PfkB family of carbohydrate kinase, amsA= amylosucrase abundant and cheap substrate sucrose and for the costeffective glycosylation of small molecules by coupling to a GT (Zervosen et al 1998;Masada et al 2007;Son et al 2009;Terasaka et al 2012;Bungaruang et al 2013;Gutmann et al 2014). Key requirements for successful application in industry predominantly consist of highly active, (thermo) stable enzymes, and a high number of regeneration cycles for UDP (De Bruyn et al 2015). To this end, SuSyAc could be a novel promising alternative to the plant SuSy's currently used.…”

Section: Industrial Applicationsmentioning

confidence: 98%

Identification of sucrose synthase in nonphotosynthetic bacteria and characterization of the recombinant enzymes

Diricks

Bruyn

Daele

et al. 2015

Appl Microbiol Biotechnol

Self Cite

View full text Add to dashboard Cite

Sucrose synthase (SuSy) catalyzes the reversible conversion of sucrose and a nucleoside diphosphate into fructose and nucleotide (NDP)-glucose. To date, only SuSy's from plants and cyanobacteria, both photosynthetic organisms, have been characterized. Here, four prokaryotic SuSy enzymes from the nonphotosynthetic organisms Nitrosomonas Europaea (SuSyNe), Acidithiobacillus caldus (SuSyAc), Denitrovibrio acetiphilus (SusyDa), and Melioribacter roseus (SuSyMr) were recombinantly expressed in Escherichia coli and thoroughly characterized. The purified enzymes were found to display high-temperature optima (up to 80 °C), high activities (up to 125 U/mg), and high thermostability (up to 15 min at 60 °C). Furthermore, SuSyAc, SuSyNe, and SuSyDa showed a clear preference for ADP as nucleotide, as opposed to plant SuSy's which prefer UDP. A structural and mutational analysis was performed to elucidate the difference in NDP preference between eukaryotic and prokaryotic SuSy's. Finally, the physiological relevance of this enzyme specificity is discussed in the context of metabolic pathways and genomic organization.

show abstract

Section: Industrial Applicationsmentioning

confidence: 98%

Identification of sucrose synthase in nonphotosynthetic bacteria and characterization of the recombinant enzymes

Diricks

Bruyn

Daele

et al. 2015

Appl Microbiol Biotechnol

Self Cite

View full text Add to dashboard Cite

show abstract

“…[55][56][57][58][59] Anomeric sugar kinases with broad substrate specificity are therefore useful tools for increasing the range of available sugar phosphates. Lavine et al and Dey found that galactokinases from T. thermophile and V. faba, respectively, can accept 2-deoxy-ᴅ-galactose and galacatosamine as substrates in addition to ᴅ-galactose.…”

Section: Substrate Specificitymentioning

confidence: 99%

Functional analysis of anomeric sugar kinases

Conway

Voglmeir

2016

Carbohydrate Research

View full text Add to dashboard Cite

“…In terms of UGTs, they have emerged as promising catalyst for UDP-sugar based glycosylation focusing on small molecules like salidroside [4]. Although many UGTs have been cloned and expressed in E. coli glycoside production remains hampered by poor expression of plant genes in this prokaryote host [11].…”

Section: Discussionmentioning

confidence: 99%

“…The majority of glycosylation reactions in nature are mediated by glycosyltransferases, which comprise a large and divergent polyphyletic multigene superfamily [1, 2]. In plants, glycosylation catalysed by uridine diphosphate (UDP) glycosyltransferases (UGTs) [2–4] is involved in the synthesis of many secondary metabolites. Studies on the recombinant expression of UGTs have resulted in the production of various plant secondary metabolites as glycosides [2, 5, 6].…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Expression of Codon-Optimized Plant GlycosyltransferaseUGT72B14inEscherichia coliEnhances Salidroside Production

Xue

Guo

et al. 2016

BioMed Research International

View full text Add to dashboard Cite

Salidroside, a plant secondary metabolite in Rhodiola, has been demonstrated to have several adaptogenic properties as a medicinal herb. Due to the limitation of plant source, microbial production of salidroside by expression of plant uridine diphosphate glycosyltransferase (UGT) is promising. However, glycoside production usually remains hampered by poor expression of plant UGTs in microorganisms. Herein, we achieved salidroside production by expression of Rhodiola UGT72B14 in Escherichia coli (E. coli) and codon optimization was accordingly applied. UGT72B14 expression was optimized by changing 278 nucleotides and decreasing the G+C content to 51.05% without altering the amino acid sequence. The effect of codon optimization on UGT72B14 catalysis for salidroside production was assessed both in vitro and in vivo. In vitro, salidroside production by codon-optimized UGT72B14 is enhanced because of a significantly improved protein yield (increased by 4.8-fold) and an equivalently high activity as demonstrated by similar kinetic parameters (K M and V max), compared to that by wild-type protein. In vivo, both batch and fed-batch cultivation using the codon-optimized gene resulted in a significant increase in salidroside production, which was up to 6.7 mg/L increasing 3.2-fold over the wild-type UGT72B14.

show abstract

Biotechnological advances in UDP-sugar based glycosylation of small molecules

Cited by 136 publications

References 213 publications

Identification of sucrose synthase in nonphotosynthetic bacteria and characterization of the recombinant enzymes

Identification of sucrose synthase in nonphotosynthetic bacteria and characterization of the recombinant enzymes

Functional analysis of anomeric sugar kinases

Expression of Codon-Optimized Plant GlycosyltransferaseUGT72B14inEscherichia coliEnhances Salidroside Production

Contact Info

Product

Resources

About