Bacterial cellulose nanocrystals obtained through enzymatic and acidic routes: A comparative study of their main properties and in vitro biological responses.

Cellulose nanocrystals (CNCs) are crystalline domains isolated from cellulosic fibers. They have been utilized in a wide range of applications, such as reinforcing fillers, antibacterial agents and manufacturing of biosensors. Whitin this context, the aim of this work was to obtain and analyze CNCs extracted from bacterial nanocellulose (BNC) using two distinct methods combined with milling pre-treatment: an acidic hydrolysis using 64 % sulfuric acid and an enzymatic hydrolysis using a commercial cellulase enzyme mixture. The CNCs obtained from the enzymatic route (e-CNCs) were observed to be spherical nanoparticles with diameter of 56 ± 11 nm. In contrast, the CNCs from the acid hydrolysis (a-CNCs) appeared as needle-shaped nanoparticles with a high aspect ratio with lengths/widths of 158 ± 64 nm/11 ± 2 nm. The surface zeta potential (ZP) of the a-CNCs was -30,8 mV, whereas the e-CNCs has a potential of +2.70 ± 3.32 mV, indicating that a-CNCs consisted of negatively charged particles with higher stability in solution. Although the acidic route resulted in nanocrystals with a slightly higher crystallinity index compared to the enzymatic route, e-CNCs was found to be more thermally stable than BNC and a-CNCs. Here, we also confirmed the safety of a-CNCs and e-CNCs using L929 cell line. Lastly, this article describes two different CNCs synthesis approaches that leads to the formation of nanoparticles with different dimensions, morphology and unique physicochemical properties. To the best of our knowledge, this is the first study to yield spherical nanoparticles as a result of BNC enzymatic treatment.

Xylanase increases the selectivity of the enzymatic hydrolysis with endoglucanase to produce cellulose nanocrystals with improved properties.

Enzyme-mediated isolation of cellulose nanocrystals (CNCs) is a promising environment friendly method with expected lower capital and operating expenditures compared to traditional processes. However, it is still poorly understood. In this study, an endoxylanase was applied as accessory enzyme to assess its potential to increase the selectivity of an endoglucanase during cellulose hydrolysis to isolate CNCs with improved properties. Only combinations of the enzymes with xylanase activity equal to or higher than the endoglucanase activity resulted in CNCs with improved properties (i.e., crystallinity, thermostability, uniformity, suspension stability and aspect ratio). The beneficial effects of the accessory enzyme are related to its hydrolytic (xylan and cellulose hydrolysis) and non-hydrolytic action (swelling of cellulose fibers and fiber porosity) and on the ratio of the enzymes, which in turn allows to tailor the properties of the CNCs. In conclusion, compared to the traditional sulfuric acid hydrolysis method, accessory enzymes help to isolate cellulose nanomaterials with improved and customized (sizes, aspect ratio and morphology) properties that may allow for new applications.