Development and Characterization of a Tunable Metal-Organic Framework (MOF) for the Synthesis of a Rare Sugar D-Tagatose.

D-tagatose is a valuable rare sugar with potential health benefits such as antiobesity, low-calorie, prebiotic, and anticancer. However, its production is mainly depending on chemical or enzymatic catalysis. Herein, a cobalt-based metal-organic framework (MOF) was developed at room temperature in an aqueous system using a self-assembly method. The L-arabinose isomerase (L-AI) was immobilized into this unique MOF by an in situ encapsulation process. The morphology and structural aspects of the MOF preparations were characterized by different analytical techniques such as scanning electron microscopy (SEM), energy dispersive X-ray spectroscopy (EDX), confocal laser scanning microscopy (CLSM), Fourier transform infrared spectroscopy (FT-IR), and X-Ray diffraction (XRD). Moreover, thermogravimetric analysis (TGA) suggested the high thermal stability of the L-AI@MOF. Significantly, the immobilized catalyst exhibited enhanced catalytic efficiency (kcat/Km) of 3.22 mM-1 s-1 and improved turnover number (kcat) of 57.32 s-1. The L-AI@MOF efficiently catalyzes the synthesis of D-tagatose from D-galactose up to the equilibrium level (~ 50%) of isomerization in heterogeneous catalysis. Interestingly, L-AI@MOF was found stable and reusable for more than five cycles without the requirement of additional metal ions during catalysis. Thus, L-AI stabilized in the MOF system demonstrated a higher catalytic activity and potential guidance for the sustainable synthesis of rare sugar D-tagatose.

Robust nano-enzyme conjugates for the sustainable synthesis of a rare sugar D-tagatose.

Aim of present study was to develop biological catalysts of L-arabinose isomerase (L-AI) by immobilizing on four different supports such as multiwalled carbon nanotube (MWCNT), graphene oxide (GOx), Santa Barbara Amorphous (SBA-15) and mobile composite matter (MCM-41). Also, comparative analysis of the developed catalysts was performed to evolve the best in terms of transformation efficiency for D-tagatose production. The developed nano-enzyme conjugates (NECs) were characterized using the high resolution transmission electron microscopy (HR-TEM) and elemental analysis was performed by energy dispersive X-ray spectroscopy (EDS). The functional groups were investigated by Fourier transform infra red spectroscopy. Also, the thermo gravimetric analysis (TGA) was employed to plot a thermal degradation weight loss profile of NECs. The conjugated L-AI with MWCNT and GOx were found to be more promising immobilized catalysts due to their ability to provide more surface area. Conversion of D-Galactose to D-Tagatose at moderate temperature and pH was observed to attain the equilibrium level of transformation (~50%). On the contrary, NECs prepared using SBA-15 and MCM-41 as support matrix were unable to reach the equilibrium level of conversion. Additionally, the developed NECs were suitable for reuse in multiple batch cycles. Thus, promising nanotechnology coupled with biocatalysis made the transformation of D-Galactose into D-tagatose more economically sustainable.

Metal-based micro-composite of L-arabinose isomerase and L-ribose isomerase for the sustainable synthesis of L-ribose and D-talose.

The biocatalysts are broadly explored in the biological transformation processes. The enzyme cascade catalysis involves various catalytic activities in a sequential process to produce the desired product including the formation of reaction intermediates. Enzyme immobilization is a method in which enzymes are confined within a support or matrix either physically or chemically to enhance their relative stability and catalytic activity in the enzyme cascade catalysis. In view of this, L-arabinose isomerase (L-AI) and L-ribose isomerase (L-RI) were immobilized on zeolite based metal framework as a micro-composite construct (DEMC@L-AI+L-RI) using linker, and metal ions. Such immobilization could be of great significance and provide several advantages like mesoporous surface for enzyme adsorption, desirable functionality in the production of products in enzyme cascade reaction, high storage stability and enhanced recyclability. The developed DEMC@L-AI+L-RI was characterized using SEM, FTIR, CLSM and TGA. The immobilization yield was 32% and loading of enzyme was 22% on the surface of micro-composite. The DEMC@L-AI+L-RI showed relatively stable catalytic activity at pH 5-6 and temperature 40 °C. The catalytic efficiency (kcat/Km) of both the enzymes was increased by 1.5-fold after immobilization. With the immobilized biocatalyst, bioconversion of L-arabinose to L-ribose was 22.6% and D-galactose to D-talose was 15.2%. The reusability of developed biocatalyst for more than six cycles was observed for more than 50% yield of the sugars. The conversion of biomass sugars from beetroot and onion waste residues was 20% and 14% to produce ribose and talose, respectively.

Recovery of nanosized silica and lignin from sugarcane bagasse waste and their engineering in fabrication of composite membrane for water purification.

Environmental benign catalytic process was developed for the valorisation of sugarcane bagasse into functional nanomaterials. Bagasse saccharification was carried out with an acid catalyst (H2SO4 0.5%, wt/wt) to separate sugars after pre-treatment of biomass with ethanol. Subsequently, a combination of peroxide and base (0.5% H2O2, wt/wt and 1% NaOH, wt/wt) was stacked to concurrently synthesise SiO2 (35 nm with 5.65% yield) and lignin (20 nm with 10.15% yield) from bagasse slurry. In the final step, precipitation using catalyst was completed to separate highly pure functional materials in powdered form. Zeta potential (ζ) of the synthesised materials was found to be - 35.6 mV for SiO2 and - 13.1 mV for lignin. Obtained silica and lignin nanomaterials were used in the fabrication of strong as well as flexible functional membrane for purification of solute particles and gases. The adsorption/desorption curve of the developed functional membrane showed type II isotherm with a H3 hysteresis loop. The observed Brunauer-Emmett-Teller surface area of the membrane was 400.3 m2/g. The pore size and pore volume as recorded by Barrett-Joyner-Halenda method was 25.5 nm and 0.624 cm3/g, respectively. Hence, the developed simple and sustainable process could be highly suitable for filtration of contaminated water and air purification.

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.

A dual-enzyme metal-organic hybrid crystal was constructed through self-assembling of manganese phosphate embedded with ß-galactosidase and L-arabinose isomerase for facile synthesis of rare sugar D-tagatose. The synthesized crystal-like hierarchical system (MnHC@ß-Gal+L-AI) was extensively characterized for structural features and catalytic reactions. The results indicated that upon immobilization onto the hybrid crystal, the activity of ß-galactosidase and L-arabinose iomerase was enhanced by a factor of 1.6- and 1.5-fold, respectively. The developed MnHC@ß-Gal+L-AI exhibited excellent efficiency with a net equilibrium level conversion of low-cost substrate whey lactose (100%) into D-glucose (∼50%), D-galactose (∼25%), and D-tagatose (∼25%). In addition, the fabricated hybrid crystals displayed cofactor regeneration ability. Therefore, the developed hybrid system was observed to be efficiently reused more than 5 times in a batch level conversion. Hence, the developed dual-enzyme-based hybrid crystal provides a platform for direct transformation of whey lactose into rare sugar D-tagatose.

Xylanase from Acinetobacter pittii MASK 25 and developed magnetic cross-linked xylanase aggregate produce predominantly xylopentose and xylohexose from agro biomass.

Most of the chemical and biochemical processes used for the de-polymerization of structural polymers of lignocellulosic biomass are environment unfriendly and costly. Here an efficient process based on xylanase, produced by Acinetobacter pittii MASK25 (MTCC 25132), hydrolysis of only physically treated rice straw and corn cob has been developed for the production of xylooligosaccharides. Bacterial strain isolated from soil was found to produce maximum xylanase at 30°C and pH 7. While the optimum temperature and pH of xylanase were characterized as 40°C and 5. Process was further improved by developing magnetic-xylanase CLEA. Crude xylanase and magnetic-xylanase CLEA could convert respectively more than 45% and 60% xylan of the powdered rice straw and corn cob into xylooligosaccharides. Interestingly, hydrolysis by both types of enzymatic forms was found to produce predominantly xylopentose and xylohexose. Hence, the process is environment friendly and the predominant production of xylopentose and xylohexose could find unique prebiotic applications.