We demonstrated the nano-immobilization of fungal enzymes through their encapsulation in cellulose nanoparticles (CNPs). An extracellular enzyme cocktail (a mixture of amylase, protease, lipase, and cellulose) was produced from Aspergillus niger and Phanerochaete chrysosporium through submerged fermentation. The process of encapsulation was carried out through a microemulsion nanoprecipitation method in the presence of a lipid, a surfactant, and a co-surfactant. The morphology of CNPs was determined by field-emission scanning electron microscopy and transmission electron microscopy; CNPs were less than 100 nm in diameter. Fourier transform infrared spectroscopy (FTIR) and energy dispersive spectroscopy demonstrated the successful encapsulation of the fungal enzyme cocktail and revealed C and O as its major components. FTIR peaks of CNPs with encapsulated enzymes occurred at 3421.80, 2828.91, 1649.29, 1450.24, and 1061.61 cm⁻¹ as well as in the range of 1050–1150 cm⁻¹. Encapsulated enzymes showed excellent stability with a peak at −70.91 mV in zeta potential studies. Thermogravimetric analysis proved that the CNP-encapsulated enzymes had an initial weight loss at 250°C. The encapsulated fungal enzyme cocktail exhibited higher catalytic performance and stability than the free enzymes. The encapsulated fungal enzyme cocktail derived from A. niger at the concentration of 100 µg/mL, showed the highest amylase activity with a clear zone of 2.5 cm. Overall, the results of this research reveal the enhancement in the activity of fungal extracellular enzyme cocktail through nanoencapsulation.

中文翻译：

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真菌细胞外酶混合物在纤维素纳米颗粒中的包裹：增强酶的稳定性

我们证明了通过封装在纤维素纳米颗粒（CNPs）中的真菌酶的纳米固定化。从黑曲霉和Phanerochaete chrysosporium生产一种细胞外酶混合物（淀粉酶，蛋白酶，脂肪酶和纤维素的混合物）通过深层发酵。在脂质，表面活性剂和助表面活性剂的存在下，通过微乳液纳米沉淀法进行包囊过程。通过场发射扫描电子显微镜和透射电子显微镜确定CNP的形态。CNP的直径小于100 nm。傅里叶变换红外光谱（FTIR）和能量分散光谱证明了真菌酶混合物的成功封装，并揭示了C和O作为其主要成分。包封的酶的CNPS FTIR峰发生在3421.80，2828.91，1649.29，1450.24，和1061.61厘米^-1以及在1150至50年cm的范围内^-1。封装的酶在zeta电位研究中显示出极好的稳定性，在-70.91 mV处有一个峰。热重分析证明，CNP包埋的酶在250°C时具有初始重量损失。包封的真菌酶混合物比游离酶具有更高的催化性能和稳定性。来自黑曲霉的封装的真菌酶混合物浓度为100 µg / mL，显示出最高的淀粉酶活性，净区为2.5 cm。总体而言，这项研究的结果表明，通过纳米封装可以增强真菌细胞外酶混合物的活性。