Single‐Cell Nanoencapsulation of Saccharomyces cerevisiae by Cytocompatible Layer‐by‐Layer Assembly of Eggshell Membrane Hydrolysate and Tannic Acid

Abstract

Single‐cell nanoencapsulation (SCNE) demands cytocompatible materials and processes to ensure the maintenance of cell viability and prefers the degradation‐on‐demand and postfunctionalization of the cytoprotective shells. Although the layer‐by‐layer (LbL) method has intensively been used for SCNE, there have been few reports on the cytocompatible LbL shells that are postfunctionalizable under mild conditions. Herein, the use of nature‐derived eggshell membrane hydrolysate (ESMH) as a counter component to tannic acid (TA) for hydrogen bonding‐based LbL shell formation on Saccharomyces cerevisiae is proposed. In addition to the great cytocompatibility of the LbL process and protective capability of the ESMH/TA shell (e.g., 18‐fold increase in survival against Cu2+), the shell is postfunctionalizable, benefitting from the presence of various functional groups in the ESMH, as demonstrated by reactions with N‐hydroxysuccinimide‐ or maleimide‐conjugated fluorescent probes and bioinspired silicification. This work suggests that ESMH would be an advanced biomaterial for chemically interfacing with living cells in a controlled fashion.

Hydrogen Bonding‐Based Layer‐by‐Layer Assembly of Nature‐Derived Eggshell Membrane Hydrolysates and Coffee Melanoidins in Single‐Cell Nanoencapsulation

“…In the exploitation of food waste as biomaterials, we have previously reported the use of ESMHs and CMs in ultrathin-film formation. The LbL films of the water-soluble ESMH, a hydrolyzed form of ESM, were formed with Fe 3 + -tannic acid (TA) complex [44] or TA alone, [45] and the CM-based films were produced via coordination complex formation with Fe 3 + ions. [46] In our previous studies, we confirmed each compound's biocompatible characteristics by the SCNE of Saccharomyces cerevisiae within ESMH/TA LbL (for ESMHs) and Fe 3 + -CM complex (for CMs) shells [45,46] and showed the neuroactive property of ESMH-based surfaces, accelerating neurite outgrowth of primary hippocampal neurons.…”

“…The LbL films of the water-soluble ESMH, a hydrolyzed form of ESM, were formed with Fe 3 + -tannic acid (TA) complex [44] or TA alone, [45] and the CM-based films were produced via coordination complex formation with Fe 3 + ions. [46] In our previous studies, we confirmed each compound's biocompatible characteristics by the SCNE of Saccharomyces cerevisiae within ESMH/TA LbL (for ESMHs) and Fe 3 + -CM complex (for CMs) shells [45,46] and showed the neuroactive property of ESMH-based surfaces, accelerating neurite outgrowth of primary hippocampal neurons. [44] In this study, we focused on the utilization of both ESMHs and CMs as biocompatible LbL components, which formed HB-based thin films and shells.…”

“…The water contact angle decreased to 47.7°� 1.6 for [ESMH/CM] 10 and 44.3°� 1.7 for [ESMH/CM] 10 [ESMH] from 72.6°� 2.7 (for bare gold) because of the hydrophilic nature of ESMHs and CMs. [45,46] Furthermore, we characterized the [ESMH/CM] n films on quartz using UV-Vis spectroscopy. The ESMH and CM had a signature peak of 200 nm (π-π* electronic transition for peptide bonds) [47] and 405 nm (specific light-absorption wavelength of melanoidins), [48] respectively, which we used as a metric for film growth on quartz (Figure S1g).…”

“…The FE-SEM images indicated the successful formation of the [ESMH/CM] 7 shell on individual S. cerevisiae: the surface became rougher, composed of nanoparticulates, after SCNE (Figure 2b). To visualize the shell, we synthesized a rhodamine-linked ESMH (ESMH_TAMRA, λ emission : 575 nm) by following our reported synthetic protocol [45] and formed the [ESMH/CM] 7 [ESMH_TAMRA] shell on the cell. The confocal laser-scanning microscopy (CLSM) image clearly showed the red rings indicative of the shell's formation (Figure 2c).…”

Synthesis of a Healthy Sweetener d-Tagatose from Starch Catalyzed by Semiartificial Cell Factories

“…Importantly, the architectures significantly increase the potential of functional biomacromolecules in industrial applications that require enhanced thermal stability, tolerance to organic solvents, or extended lifetimes. Recent investigations have demonstrated that coating cells with a protective shell, such as metal–organic frameworks, − mineralized oxides, ,− and polyphenols, − has good application prospects in the biomedical field, but these cell coating methods are difficult to use in practical applications due to the cumbersome and high cost of synthetic methods in the field of biomanufacturing.…”