Department of Biochemistry, Maria Curie-Sklodowska University, Akademicka 19, 20-033 Lublin, Poland.
Department of Biochemistry, Maria Curie-Sklodowska University, Akademicka 19, 20-033 Lublin, Poland.
Fungal Biol. 2019 Dec;123(12):875-886. doi: 10.1016/j.funbio.2019.09.007. Epub 2019 Sep 24.
Cellobiose dehydrogenase (CDH, EC 1.1.99.18) is a glycoprotein having many biotechnological applications. In the present study, CDHs isolated from Phlebia lindtneri (PlCDH), Phanerochaete chrysosporium (PchCDH), Cerrena unicolor (CuCDH), and Pycnoporus sanguineus (PsCDH) were studied the first time for their ability to generate antioxidant and antimicrobial agents. The aim of the research was to evaluate the antioxidant and antimicrobial activity of systems composed of four CDHs and lactose or cellobiose as a reaction substrate. The free radical scavenging effect of free and immobilised enzymes was evaluated using the DPPH method. The lowest values of EC (10.04 ± 0.75 μg/ml) was noted for PlCDH/lactose and for PlCDH/cellobiose (12.06 ± 1.35 μg/ml). The ECvalue reached 12.6 ± 1.51 μg/ml in the case of PsCDH/lactose and 15.96 ± 1.35 for PsCDH. The CDH preparations were also effectively immobilised in alginate (the immobilisation efficiency expressed as a protein yield ranged from 61.6 to 100 %). The operational stability expressed as a scavenging effect showed the possibility of using the alginate beads 4 times. Both the free and immobilised CDHs as well as the CDH/substrate were tested against Gram-negative Escherichia coli ATCC 25922, Pseudomonas aeruginosa ATCC 27853, and Gram-positive Staphylococcus aureus ATCC 25923 bacteria. All samples, except PlCDH, were potentially effective in suppression of bacterial growth. The highest percentage of inhibition (100 %) was obtained for S. aureus bacteria using PsCDH and PchCDH with lactose as a substrate, whereas a slightly lesser effect was observed for E. coli and P. aeruginosa bacterial cells, i.e. 64.1 % and 86.5 % (PsCDH) and 94.1 % and 41.4 % (PchCDH), respectively. Furthermore, the concentrations of the reaction products (aldonic acids and hydrogen peroxide) were quantified and the surface morphology of the alginate beads was analysed using SEM visualisation.
纤维二糖脱氢酶（CDH，EC 1.1.99.18）是一种具有许多生物技术应用的糖蛋白。在本研究中，首次研究了从 Phlebia lindtneri（PlCDH）、Phanerochaete chrysosporium（PchCDH）、Cerrena unicolor（CuCDH）和 Pycnoporus sanguineus（PsCDH）中分离出的 CDH 产生抗氧化和抗菌剂的能力。研究的目的是评估由四种 CDH 和乳糖或纤维二糖作为反应底物组成的系统的抗氧化和抗菌活性。使用 DPPH 法评估游离和固定化酶的自由基清除效果。PlCDH/乳糖和 PlCDH/纤维二糖的 EC（10.04±0.75μg/ml）值最低，而 PsCDH/乳糖和 PsCDH 的 EC 值分别为 12.6±1.51μg/ml 和 15.96±1.35μg/ml。CDH 制剂也有效地固定在海藻酸钠中（固定化效率表示为蛋白产率范围为 61.6 至 100%）。以清除效果表示的操作稳定性表明，海藻酸钠珠可使用 4 次。游离和固定化的 CDH 以及 CDH/底物均针对革兰氏阴性大肠杆菌 ATCC 25922、铜绿假单胞菌 ATCC 27853 和革兰氏阳性金黄色葡萄球菌 ATCC 25923 细菌进行了测试。除 PlCDH 外，所有样品均能有效抑制细菌生长。使用 PsCDH 和 PchCDH 与乳糖作为底物时，对 S. aureus 细菌的抑制率最高（100%），而对 E. coli 和 P. aeruginosa 细菌细胞的抑制效果略低，分别为 64.1%和 86.5%（PsCDH）和 94.1%和 41.4%（PchCDH）。此外，定量了反应产物（醛酸和过氧化氢）的浓度，并使用 SEM 可视化分析了海藻酸钠珠的表面形态。
