Simulation and economic assessment of large-scale enzymatic N-acetyllactosamine manufacture

Alavijeh, Masih Karimi; Meyer, Anne S.; Gras, Sally L.; Kentish, Sandra E.

doi:10.1016/j.bej.2019.107459

Cited by 10 publications

(10 citation statements)

References 19 publications

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“…We have previously developed a simulation of a LacNAc production process using this β-galactosidase. 2 This simulation was now used to compare the amount of LacNAc that could be obtained from 20 tons of lactose derived from whey, as well as the annual cost of LacNAc production, for both the free and the immobilized enzyme. The results are shown in Table S2 in the Supporting Information, indicating the annual production of 8.5 tons LacNAc with the use of immobilized enzyme and 6.3 tons LacNAc with the use of free enzyme.…”

Section: Preliminary Economic Analysis Resultsmentioning

confidence: 99%

“…This is an important result, which shows that the immobilization technique employed in this study not only makes it possible to recover the enzyme after the reaction but also that immobilization increases yield, which can significantly influence the economy of largescale LacNAc production. 2 For further elucidation, as shown in Figure 4(C), the reaction specific productivity (also known as biocatalytic productivity rate) was calculated at two time points when the highest LacNAc concentrations were obtained in the reactions catalyzed by the free enzyme at 0.63 U•ml -1 (90 min) and the immobilized enzyme at 0.14 U•ml -1 (150 min), respectively. When the free enzyme is used, a greater amount of LacNAc can be achieved using 0.63 U•ml -1 rather than 0.14 U•ml -1 but at the expense of a reduction in the specific productivity.…”

Section: Transgalactosylation Reactionmentioning

confidence: 99%

“…The global dairy industry produces large amounts of lactose-rich by-product streams, particularly whey generated from manufacturing of cheese, casein and yoghurt. [1][2] Lactose valorization-the use of lactose in the synthesis of functional food and pharmaceutical products-can enhance the economic and environmental sustainability of the industry. 1,3 Although separation of proteins as a value-added product from whey streams has become an established process over the past 30 years, lactose valorization is still under development.…”

Section: Introductionmentioning

confidence: 99%

“…The global dairy industry produces large amounts of lactose-rich byproduct streams, particularly whey generated from manufacturing of cheese, casein, and yogurt. , Lactose valorizationthe use of lactose in the synthesis of functional food and pharmaceutical productscan enhance the economic and environmental sustainability of the industry. , Although separation of proteins as a value-added product from whey streams has become an established process over the past 30 years, lactose valorization is still under development. Large amounts of the lactose generated are not used and so regarded as low-value streams, imposing adverse environmental impacts. ,, One solution to exploit this abundant resource is to synthesize lactose-derived prebiotics, including galacto-oligosaccharides or human milk oligosaccharides, via β-galactosidase-catalyzed transgalactosylation reactions.…”

Section: Introductionmentioning

confidence: 99%

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Improving β-Galactosidase-Catalyzed Transglycosylation Yields by Cross-Linked Layer-by-Layer Enzyme Immobilization

Alavijeh

Meyer

Gras

et al. 2020

ACS Sustainable Chem. Eng.

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The biotransformation of lactose into gut-bioactive glycans catalyzed by βgalactosidase can give economic value to lactose-rich side streams generated in the food or dairy industry. Herein, we study the immobilization of the commercially used β-galactosidase from Bacillus circulans onto silica particles using an enzyme immobilization technology involving a cross-linked layer-by-layer encapsulation method. The immobilized β-galactosidase was used for the synthesis of Nacetyllactosamine (LacNAc) as an important precursor for numerous bioactive compounds and a prebiotic in itself. Techniques including molecular analysis, enzyme activity determination, secondary structure analysis, thermodynamic characterization as well as the determination of thermal and operational stability were conducted to characterize the immobilized enzyme. Changes in the activity of the enzyme after immobilization were attributed to possible changes in electrostatic, covalent and protein-protein interactions. Immobilization significantly improved enzymatic LacNAc yield compared to the free enzyme. In turn, this improved the economics and the sustainability of the process. The immobilized enzyme encapsulated in multilayer films was significantly more stable in the presence of divalent cations and its thermostability also substantially increased, with the thermal denaturation activation energy increasing from 53 kJ mol -1 to 294 kJ mol -1 . The immobilized enzyme was successfully reused in eight consecutive reaction cycles with no significant reduction in the LacNAc yield. The improved transgalactosylation yield and productivity, higher stability and reusability obtained with this immobilization method provide new opportunities for industrial applications.

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Section: Preliminary Economic Analysis Resultsmentioning

confidence: 99%

Section: Transgalactosylation Reactionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Improving β-Galactosidase-Catalyzed Transglycosylation Yields by Cross-Linked Layer-by-Layer Enzyme Immobilization

Alavijeh

Meyer

Gras

et al. 2020

ACS Sustainable Chem. Eng.

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“…The use of methanol, however, can be problematic in terms of waste treatment and storage costs due to the flammability of this solvent. 208 As an alternative to expensive chromatographic purification methods, membrane separation of galactooligosaccharides as a mixture from other low molecular weight sugars has been investigated in numerous studies. [209][210][211][212][213][214] Given a typical molecular weight range of 200-1000…”

Section: Purification Methods For Large-scale Synthesismentioning

confidence: 99%

Synthesis of N-Acetyllactosamine and N-Acetyllactosamine-Based Bioactives

Alavijeh

Meyer

Gras

et al. 2021

J. Agric. Food Chem.

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N-Acetyllactosamine (LacNAc), or more specifically β-D-galactopyranosyl-1,4-N-acetyl-Dglucosamine, is a unique acyl-amino sugar and a key structural unit in human milk oligosaccharides, an antigen component of many glycoproteins and an antiviral active component for the development of effective drugs against viruses. LacNAc is useful itself and as a basic building block for producing various bioactive oligosaccharides, notably because this synthesis may be used to add value to dairy lactose. Despite a significant amount of information in the literature on the benefits, structures and types of different LacNAc-derived oligosaccharides, knowledge about their effective synthesis for large scale production is still in its infancy. This work provides a comprehensive analysis of existing production strategies of LacNAc and important LacNAc-based structures, including sialylated LacNAc, as well as poly-and oligo-LacNAc. We conclude that direct extraction from milk is too complex, while chemical synthesis is also impractical at an industrial scale. Microbial routes have application when multiple step reactions are needed but the major route to large scale biochemical production will likely lie with enzymatic routes, particularly those using β-galactosidases (for LacNAc synthesis), sialidases (for sialylated LacNAc synthesis) and β-N-acetylhexosaminidases (for oligo-LacNAc synthesis).Glycosyltransferases, especially for the biosynthesis of extended complex LacNAc structures, could also play a major role in the future. In these cases, immobilization of the enzyme can increase stability and reduce cost. Processing parameters, such as substrate concentration and purity, acceptor to donor ratio, water activity and temperature can affect product selectivity and yield.More work is needed to optimize these reaction parameters and in the development of robust, thermally stable enzymes to facilitate commercial production of these important bioactive substances. Abbreviations: 39 β3GalT β1-3 galactosyltransferase β4GalT β1-4 galactosyltransferase CMP-NeuNAc cytidine-5'-monophosphate-5-N-acetylneuraminic acid DS'LNT α2-6-linked disialyllacto-N-tetraose DSLNnT α2-6-linked disialyllacto-N-neotetraose EPEC enteropathogenic Escherichia coli GalNAc N-acetylgalactosamine GfCoV Guinea fowl coronavirus GlcNAc N-acetylglucosamine GLP-1 glucagon-like peptide 1 HEp-2 human epithelial type 2 cells iGnT β1-3 N-acetylglucosaminyltransferase IGnT β1-6 N-acetylglucosaminyltransferase iLNO iso-Lacto-N-octaose LacNAc N-acetyllactosamine LacNAc-ITag imidazolium-tagged LacNAc LacY β-galactoside permease lacZ β-galactosidase gene lgtA β1-3 N-acetylglucosaminyltransferase gene lgtB β1-4 galactosyltransferase gene LND Lacto-N-decaose LNH Lacto-N-hexaose LNnD Lacto-N-neo-decaose LNnH Lacto-N-neo-hexaose LNnO Lacto-N-neo-octaose LNnT Lacto-N-neotetraose LNO Lacto-N-octaose LNT Lacto-N-tetraose ManNAc N-acetylmannosamine MERS-CoV Middle East respiratory syndrome coronavirus Neu5Ac N-acetylneuraminic acid oNPG ortho-nitrophenyl-β-galactoside pLNH para-Lacto-N...

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Whey Protein from Milk as a Source of Nutraceuticals

Awuchi

2022

Food and Agricultural Byproducts as Important Source of Valuable Nutraceuticals

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Simulation and economic assessment of large-scale enzymatic N-acetyllactosamine manufacture

Cited by 10 publications

References 19 publications

Improving β-Galactosidase-Catalyzed Transglycosylation Yields by Cross-Linked Layer-by-Layer Enzyme Immobilization

Improving β-Galactosidase-Catalyzed Transglycosylation Yields by Cross-Linked Layer-by-Layer Enzyme Immobilization

Synthesis of N-Acetyllactosamine and N-Acetyllactosamine-Based Bioactives

Whey Protein from Milk as a Source of Nutraceuticals

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