Large-scale purification of epilactose using a semi-preparative HPLC system

Kuschel, Beatrice; F, Riemer; Pfost, Daniel; Conrad, Jürgen; Losch, Carsten; Claaßen, Wolfgang; Beifuß, Uwe; Weiß, Jochen; Mu, Wanmeng; Stressler, Timo; Fischer, Lutz

doi:10.1007/s00217-016-2752-7

Cited by 11 publications

(4 citation statements)

References 37 publications

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“…However, the selectivity of hydrolysis by Bacillus circulans β-galactosidase and purity of product (91.1%) were relatively low (Saburi et al, 2010). Kuschel et al (2016) developed a non-food-grade method to produce epilactose; under the best conditions, the final epilactose purity was 99% with a total yield of 51%.…”

Section: Discussionmentioning

confidence: 99%

Construction of an enzymatic route using a food-grade recombinant Bacillus subtilis for the production and purification of epilactose from lactose

Chen

Yan

et al. 2018

Journal of Dairy Science

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Lactose is a main by-product in the cheese industry. Many attempts have been made to convert the lactose to high value-added products, including epilactose. Epilactose is a valuable prebiotic and can be epimerized from lactose with cellobiose 2-epimerase (CEase). The objective of the present work was to construct a food-grade recombinant Bacillus subtilis that produces CEase from Thermoanaerobacterium saccharolyticum. The CEase was expressed in B. subtilis without antibiotic resistance genes. After fermentation, the maximum volumetric activity of the fermented broth was more than 7 U/mL. The activity of the recombinant B. subtilis was increased by up to 3.7 fold after ethanol permeabilization. Then, 66.9 ± 0.7 g/L of epilactose was produced from 300 g/L of whey powder solution in 1 h with 13.3 U/mL of permeabilized biocatalyst. In addition, an enzymatic route including degradation of the lactose, yeast fermentation, and cation exchange chromatography was described to further purify the produced epilactose from lactose. Finally, epilactose with a purity >98% was produced from 300 g/L of lactose with a yield of 24.0%. In conclusion, neither antibiotics nor pathogenic bacteria were used throughout the epilactose production and purification procedure.

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Section: Discussionmentioning

confidence: 99%

Construction of an enzymatic route using a food-grade recombinant Bacillus subtilis for the production and purification of epilactose from lactose

Chen

Yan

et al. 2018

Journal of Dairy Science

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“…Moreover, the establishment of a strategy with only two steps represents a significant improvement when compared with the reported methodologies, as it significantly reduces the time and costs of the purification process. In fact, only Kuschel and collaborators presented a two-step process, but the epilactose yield was considerably low [16]. In addition, the authors used crystallization and semi-preparative chromatography, which are more complex and expensive methodologies than the ones herein described.…”

Section: Epilactose Purification Strategymentioning

confidence: 99%

“…Therefore, techniques based on the molecular size, but also enzymatic approaches, can be very tricky due to the similarity of the molecules, and need to be carefully explored. Most purification approaches rely on the use of high-performance liquid chromatography (HPLC) and ion-exchange resins which usually includes a high number of steps and results in low yields [7,16,20]. Hence, this study was focused on the development of a suitable and simple strategy to purify epilactose aiming not only to reduce the number of steps, but also to enhance the yield.…”

Section: Introductionmentioning

confidence: 99%

Two-Step Purification of Epilactose Produced by Cellobiose 2-Epimerase from Caldicellulosiruptor Saccharolyticus

Cardoso¹,

Fernandes²,

Pinheiro³

et al. 2022

SSRN Journal

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“…[24][25][26] Semi-preparative highperformance liquid chromatography (semi-preparative HPLC), as a classical chromatography method is also widely applied in the separation and purification of natural products. 27 Moreover, the combination of CCC of liquid-liquid system and semi-preparative HPLC of liquidsolid system has complementary advantages for separating bioactive compounds from crude extracts of C. arietinum L., [28][29][30] demonstrating its promising potential for industrial applications. 31 Herein, we screened target compounds by employing the UF assay based on receptor-ligand affinity, which was expected to screen potential XOD inhibitors out.…”

mentioning

confidence: 99%

In‐depth analysis of the xanthine oxidase inhibitors of Cicer arietinum L.‐based receptor–ligand affinity coupled with complex chromatography

Hou

Niu

Liu

et al. 2023

Phytochemical Analysis

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IntroductionCicer arietinum L. is the choice of health food for people with diabetes, hypertension, and hyperlipidemia. As an essential source of high‐nutrition legumes, it is also an important source of dietary isoflavones.ObjectivesIn order to improve the preparation efficiency of natural plants, a rapid biological activity screening and preparation of xanthine oxidase inhibitors from C. arietinum L. was established.MethodsXanthine oxidase (XOD) inhibitors were rapidly screened using ultrafiltration liquid chromatography–mass spectrometry (UF‐LC–MS) based on receptor–ligand affinity. The change in XOD activity was evaluated by enzymatic reaction kinetics measurement. The potential bioactive compounds were verified through molecular docking. In addition, the biological activity of ligands screened was separated and purified by complex chromatography. The structures of the compounds were identified by nuclear magnetic resonance spectroscopy.ResultsThree active ingredients, namely daidzin, daidzein, calycosin with XOD binding affinities were identified and isolated from the raw plant materials via semi‐preparative high‐performance liquid chromatography (HPLC), 0–60 min, 5–50% B and countercurrent chromatography (CCC) (ethyl acetate/acetic acid/water [5:0.8:10, v/v/v]).ConclusionThis study will help to elucidate the mechanisms of action of natural plants of interest at the molecular level and could also provide more opportunities for the discovery and development of new nutritional value from other natural resources.

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Large-scale purification of epilactose using a semi-preparative HPLC system

Cited by 11 publications

References 37 publications

Construction of an enzymatic route using a food-grade recombinant Bacillus subtilis for the production and purification of epilactose from lactose

Construction of an enzymatic route using a food-grade recombinant Bacillus subtilis for the production and purification of epilactose from lactose

Two-Step Purification of Epilactose Produced by Cellobiose 2-Epimerase from Caldicellulosiruptor Saccharolyticus

In‐depth analysis of the xanthine oxidase inhibitors of Cicer arietinum L.‐based receptor–ligand affinity coupled with complex chromatography

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