In this study, the formation of protein microspheres through lysosomal enzyme-assisted biomineralized crystallization was demonstrated. Spherical micro-sized hybrid CaCO₃ constructs were synthesized and characterized using field-emission scanning electron microscopy equipped with energy-dispersive X-ray spectroscopy, X-ray diffraction, Fourier-transform infrared spectroscopy, and particle size analysis. Additionally, parameters such as the Brunauer–Emmett–Teller surface area and single-point total pore volume, and adsorption/desorption analysis were used to investigate the mesoporous properties, which are advantageous for lysosomal enzyme (LE) loading. A LE can be used as an organic template, not only as a morphological controller but also for entrapping LE during the crystallization pathway. The hybrid protein microspheres accommodated 2.3 mg of LE with a 57% encapsulation efficiency and 5.1 wt% loading. The peroxidase activity of the microspheres was calculated and found to be approximately 0.0238 mM⁻¹ min⁻¹. pH-responsive release of the LE from CaCO₃ was observed, suggesting potential biomedical and cosmetic applications in acidic environments. The hybrid LE microsphere treatment significantly alleviated melanin production in a dose-dependent manner and further downregulated the mRNA expression of MITF, tyrosinase, TYRP-1, and TYRP-2. These results indicate skin-whitening effects by inhibiting melanin without inducing cytotoxicity. The data provide the first evidence of the potential use of a LE for obtaining hybrid minerals and the effectiveness of biomineralization-based sustainable delivery of enzyme-based vehicles based on organelle-extract-assisted biomineralization.

中文翻译：

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杂化蛋白微球及其响应释放行为和对黑色素合成的抑制作用

在这项研究中，证明了通过溶酶体酶辅助生物矿化结晶形成蛋白质微球。使用配备能量色散X射线光谱、X射线衍射、傅里叶变换红外光谱和粒度分析的场发射扫描电子显微镜合成并表征了球形微米级杂化CaCO ₃结构。此外，还使用 ​​Brunauer-Emmett-Teller 表面积和单点总孔体积等参数以及吸附/解吸分析来研究介孔特性，这有利于溶酶体酶（LE）的负载。 LE 可以用作有机模板，不仅作为形态控制器，还可以在结晶途径中捕获 LE。混合蛋白微球可容纳 2.3 mg LE，封装效率为 57%，负载量为 5.1 wt%。计算微球的过氧化物酶活性并发现其约为0.0238 mM ^-1 min ^-1。观察到LE 从 CaCO ₃中的 pH 响应释放，表明在酸性环境中具有潜在的生物医学和化妆品应用。混合 LE 微球治疗以剂量依赖性方式显着减轻黑色素产生，并进一步下调 MITF、酪氨酸酶、TYRP-1 和 TYRP-2 的 mRNA 表达。这些结果表明通过抑制黑色素而不诱导细胞毒性来达到美白皮肤的效果。这些数据提供了第一个证据，证明 LE 在获取混合矿物质方面的潜在用途，以及基于细胞器提取物辅助生物矿化的酶载体的生物矿化可持续输送的有效性。