Combined cross-linked enzyme aggregates (combi-CLEAs) of xylanase, cellulase and β-1,3-glucanase.

Staphylococcus sp. K2204, a bacterial isolate, was employed in this work to decolorize Remazol Brilliant Blue R (RBBR), which belongs to the anthraquinone class of textile dye. Staphylococcus sp. K2204 biodegraded 100 mg/L RBBR at 37°C under static condition with the help of extracellular laccase and peroxidases. The products of RBBR degradation were characterized using analytical tools including mass spectral technique. The phytotoxicity tests evaluated the toxicity of RBBR and the products of biodegradation. The research outlined here is the first attempt to utilize Staphylococcus sp. K2204 for remediating the wastewater containing anthraquinone textile dye.

Enzyme cocktails of reusable, highly stable cellulolytic enzymes play an inevitable role in bioconversion of biomass to biofuels economically. Cellulase, xylanase and β-1,3-glucanase bound silica-amine functionalized iron oxide magnetic nanoparticles (ISN-CLEAs) were prepared and used as the biocatalyst for the depolymerization of cellulosic biomass into monomeric sugar in the present study. The FeO-NPs and FeO@SiO-NH-NPs and ISN-CLEAs had an average hydrodynamic size of 82.2, 86.4, and 976.9 nm, respectively, which was confirmed by dynamic light scattering (DLS). About 97% of protein binding was achieved with 135 mM glutaraldehyde at 10 h of cross-linking time and successful binding was confirmed by Fourier transform infrared spectroscopy (FTIR). The ISN-CLEAs exhibited the highest thermal stability of 95% at 50 °C for 2 h and retained extended storage stability of 97% compared to 60% of its free counterpart. Besides, cross-linking allowed ISN-CLEAs reuse for at least eight consecutive cycles retaining over 70% of its initial activity. ISN-CLEAs exhibited approximately 15% increase in carbohydrate digestibility on sugar cane bagasse and eucalyptus pulp than the free enzyme.

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A strain of Bacillus amyloliquefaciens KUB29 was identified by 16S ribosomal RNA sequencing (Genbank: MF772779.1). Production of thermostable protease, amylase and lipase were done by the isolated strain. The produced enzymes were partially purified by ammonium precipitation followed by dialysis process. Protease and lipase enzymes are effectively used in bio-oil extraction from proteinaceous sample followed by transesterification to produce methyl ester. Amylase enzyme is widely used in food and laundry industry. The produced enzymes are active at thermophilic condition of 55 °C. Use of these enzymes in biofuel production process will make the process cleaner and greener.

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