Chitosan-based hybrid immobilization in bienzymatic reactions and its application to the production of laminaribiose

Müller, Clarissa; Hartig, Dave; Vorländer, Karl; Sass, Ann-Cathérine; Scholl, Stephan; Jördening, Hans‐Joachim

doi:10.1007/s00449-017-1797-8

Cited by 17 publications

(21 citation statements)

References 14 publications

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“…LP and SP were immobilized through a hybrid-immobilization procedure developed in previous studies following a two-pearl concept [6,7,10]. In this procedure, a covalent linkage on Sepabeads (EC-EP/S with 0.1-0.3 mm particle size) was performed first.…”

Section: Immobilized Enzymesmentioning

confidence: 99%

“…Laminaribiose was produced in batch experiments to gain a representative mixture of sugars adsorbed on zeolite BEA, see [7] for details. The loaded zeolites were then washed and desorbed with DSP A (two washing steps, two pre-desorption steps and one final desorption) to evaluate the influence of the washing buffer.…”

Section: Optimization Of Washing and Desorption Proceduresmentioning

confidence: 99%

“…More specifically, GI as a main enzyme in production of high fructose corn syrup (HFCS) is used at much higher temperatures. Although diffusion in chitosan is significantly slower than free diffusion [8], the impact on overall reaction rate is small [9] and thermal stability of chitosan-based hybrid immobilizates is increased by a factor of 10 compared to native enzymes [7]. But glucose is needed for start of the reaction and kinetics requirements and for that reason, glucose is added at the beginning of the reaction [5].…”

mentioning

confidence: 99%

“…Furthermore, the required amount of glucose for this reaction system (which was initially supposed to be produced by GI) is produced via partial hydrolysis action of SP. Hence, in a previous work this biocatalyst system was used to set-up an enzymatic synthesis in a continuous packed bed production system with an operational time of 10 days [10] but higher productivity in continuous operation comes with a lower titer and purity of laminaribiose compared to batch mode [7]. Therefore, a bienzymatic reaction system using LP and SP immobilized on Sepabeads and encapsulated in chitosan with sucrose and glucose as substrates was applied to produce laminaribiose under mild conditions [6,7].…”

mentioning

confidence: 99%

“…But glucose is needed for start of the reaction and kinetics requirements and for that reason, glucose is added at the beginning of the reaction [5]. Therefore, a bienzymatic reaction system using LP and SP immobilized on Sepabeads and encapsulated in chitosan with sucrose and glucose as substrates was applied to produce laminaribiose under mild conditions [6,7]. Although diffusion in chitosan is significantly slower than free diffusion [8], the impact on overall reaction rate is small [9] and thermal stability of chitosan-based hybrid immobilizates is increased by a factor of 10 compared to native enzymes [7].…”

mentioning

confidence: 99%

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Continuous enzymatic production and adsorption of laminaribiose in packed bed reactors

Abi

Hartig

Vorländer

et al. 2018

Engineering in Life Sciences

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Bienzymatic production of laminaribiose from sucrose and glucose was combined with adsorption on zeolite BEA to introduce a first capture and purification step. Downstream processing including washing and desorption steps was characterized and optimized on a milliliter scale in batch mode. Results were then transferred to a packed bed system for enzymatic production and adsorption where the influence of adsorbent particle diameter on purity and productivity was evaluated. Finally, a continuous enzymatic production of laminaribiose was conducted over 10 days. The subsequent downstream processing of the loaded zeolites led to purities of over 0.5 gLaminaribiose gsugar−1 in the desorbate with a total productivity of 5.6 mgLaminaribiose Lenzyme bed−1 h−1 without the use of recycles.

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Section: Immobilized Enzymesmentioning

confidence: 99%

Section: Optimization Of Washing and Desorption Proceduresmentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

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Continuous enzymatic production and adsorption of laminaribiose in packed bed reactors

Abi

Hartig

Vorländer

et al. 2018

Engineering in Life Sciences

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The Construction of an In Vitro Synthetic Enzymatic Biosystem that Facilitates Laminaribiose Biosynthesis from Maltodextrin and Glucose

Sun

Wei

You

2019

Biotechnology Journal

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Laminaribiose is a reducing disaccharide linked by a β‐1,3 glycosidic bond; it is also a precursor for building blocks in the pharmaceutical industry, a powerful germinating agent and antiseptic, as well as a potential prebiotic. In this study, an in vitro enzymatic biosystem composed of α‐glucan phosphorylase, laminaribiose phosphorylase, isoamylase, and 4‐glucanotransferase is designed for the one‐pot synthesis of laminaribiose from low‐cost maltodextrin and glucose. Through condition optimization, 51 mM laminaribiose is produced from 10 g L−1 maltodextrin (55.5 mM glucose equivalent) and 90 mM glucose. The product yield based on maltodextrin is 91.9%. To investigate the industrial potential of this in vitro enzymatic biosystem, the production of laminaribiose from high concentrations of substrates is also examined, and 179 mM laminaribiose is produced from 50 g L−1 of maltodextrin and 450 mM glucose. This in vitro enzymatic biosystem comprised of thermophilic enzymes can drastically decrease the manufacturing cost of laminaribiose and provide a green method for the production of other disaccharides using phosphorylases.

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Three‐Enzyme Phosphorylase Cascade for Integrated Production of Short‐Chain Cellodextrins

Zhong

Nidetzky

2019

Biotechnology Journal

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Cellodextrins are linear β‐1,4‐gluco‐oligosaccharides that are soluble in water up to a degree of polymerization (DP) of ≈6. Soluble cellodextrins have promising applications as nutritional ingredients. A DP‐controlled, bottom‐up synthesis from expedient substrates is desired for their bulk production. Here, a three‐enzyme glycoside phosphorylase cascade is developed for the conversion of sucrose and glucose into short‐chain (soluble) cellodextrins (DP range 3–6). The cascade reaction involves iterative β‐1,4‐glucosylation of glucose from α‐glucose 1‐phosphate (αGlc1‐P) donor that is formed in situ from sucrose and phosphate. With final concentration and yield of the soluble cellodextrins set as targets for biocatalytic synthesis, three major factors of reaction efficiency are identified and partly optimized: the ratio of enzyme activity, the ratio of sucrose and glucose, and the phosphate concentration used. The efficient use of the phosphate/αGlc1‐P shuttle for cellodextrin production is demonstrated and the soluble product at 40 g L−1 is obtained under near‐complete utilization of the donor substrate offered (88 mol% from 200 mm sucrose). The productivity is 16 g (L h)−1. Through a simple two‐step route, the soluble cellodextrins are recovered from the reaction mixture in ≥95% purity and ≈92% yield. Overall, this study provides the basis for their integrated production.

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Chitosan-based hybrid immobilization in bienzymatic reactions and its application to the production of laminaribiose

Cited by 17 publications

References 14 publications

Continuous enzymatic production and adsorption of laminaribiose in packed bed reactors

Continuous enzymatic production and adsorption of laminaribiose in packed bed reactors

The Construction of an In Vitro Synthetic Enzymatic Biosystem that Facilitates Laminaribiose Biosynthesis from Maltodextrin and Glucose

Three‐Enzyme Phosphorylase Cascade for Integrated Production of Short‐Chain Cellodextrins

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