Self-Assembled Hybrid Nanoflowers of Manganese Phosphate and l-Arabinose Isomerase: A Stable and Recyclable Nanobiocatalyst for Equilibrium Level Conversion of d-Galactose to d-Tagatose

Abstract

We report on the synthesis and characterization of a novel hybrid nanoflower of manganese and l-arabinose isomerase and its application in synthesis of d-tagatose, a rare sugar of high commercial value. An open reading frame of 1425 base pairs from Lactobacillus sakai was used to synthesis recombinant l-arabinose isomerase of 474 amino acids in E. coli. A hierarchical flower-like spherical structure with several nanopetals was self-assembled by using purified recombinant l-arabinose isomerase as the organic component and manganese phosphate as the inorganic component. The hybrid nanoflower was characterized by scanning electron microscopy, high-resolution transmission electron microscopy, confocal laser scanning microscopy, Fourier-transform infrared spectroscopy, X-ray diffraction, and energy-dispersive X-ray spectroscopy. Circular dichroism documented no change in structural properties of l-arabinose isomerase assembled in the hybrid nanoflower. Kinetic parameters of l-arabinose isomerase were improved in the hybrid nanoflower. l-Arabinose isomerase in the manganese hybrid nanoflower was found to convert d-galactose to d-tagatose with a conversion rate of ∼50%, without an addition of manganese in the reaction mixture. The hybrid nanoflowers exhibited excellent reusability and reproducibility during reaction cycle analysis. Hence, the developed manganese hybrid nanoflowers of l-arabinose isomerase shows promise for commercial production of the rare sugar d-tagatose.

Immobilization of transaminase from Bacillus licheniformis on copper phosphate nanoflowers and its potential application in the kinetic resolution of RS-α-methyl benzyl amine

“…The reusability can be improved by adding some stabilizer during nanoflower synthesis which is known to reduce leaching of enzymes and also by the careful washings of reaction mixture during recycling. These results are in consistent with previous studies carried out by other researchers in which they immobilized different enzymes for the kinetic resolution of the racemic mixture (Rai et al 2018;Soni et al 2018;Dwivedee et al 2018).…”

“…Hybrid organic-inorganic NFs is a promising breakthrough in enzyme immobilization, which exhibits enhanced enzymatic activity and stability as compared to free enzymes, which may be attributed to the confinement of the enzyme in the core of the NFs and high-surface area (Ge et al 2012;Lee et al 2015;Li et al 2021). The combined functionalities of the protein and the inorganic material of the hybrid NFs, enables its application in biosensors (Gao et al 2020;Zhu et al 2017Zhu et al , 2018, biofuel cells (Maleki et al 2019), and in biocatalysis (Rai et al 2018).The most peculiar facet of the protein-inorganic hybrid NFs is synthesized by the conventional incubation method (reaction of mixture after three days, at room temperature), which significantly reduces their use in actual application. Nanoflower synthesis through the ultrafast sonochemical method accomplishes the limitation of the conventional method (Batule et al 2015;Dwivedee et al 2018).…”

Dual-Enzyme Metal Hybrid Crystal for Direct Transformation of Whey Lactose into a High-Value Rare Sugar D-Tagatose: Synthesis, Characterization, and a Sustainable Process

“…This weight loss could be due to pyrolytic decomposition of protein molecules encapsulated in the nanocrystals. A similar weight loss profile has earlier been documented for the calcium phosphate nanoflower with α-amylase and manganese phosphate with L-arabinose isomerase …”

“…Similar flower-like crystal structures have also been reported earlier using calcium phosphate and copper phosphate as the inorganic component. However, recently, we developed a unique nanoflower structure of manganese phosphate and L-arabinose isomerase for efficient and recyclable use in the conversion reaction of D-galactose to D-tagatose . In the present study, similar crystal structures of manganese phosphate were developed wherein two enzyme proteins β-galactosidase (β-Gal) and L-arabinose isomerase (L-AI) were embedded for direct transformation of lactose or whey lactose into D-tagatose, a rare sugar of high value.…”

Encapsulation of Mannose-6-phosphate Isomerase in Yeast Spores and Its Application in l-Ribose Production

“…Currently, rare sugars have become increasingly attractive and demonstrated great application prospects in the fields of food, medicine, and health care. − The “Izumoring strategy”, which is based on the enzymatic interconversions between two or more sugars, is very useful and widely adopted for rare sugar synthesis − Moreover, l -ribose has also been employed as a building block for the synthesis of glycoconjugates or L-aptamers …”