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L-asparaginase, a key enzyme in amino acid metabolism, is widely used in the food and pharmaceutical industries. The gut microbiota and its product L-asparaginase are closely associated with host health and diseases. [Objective] This study aims to acquire a novel L-asparaginase gene from gut microbiota and explore its enzymatic characteristics and potential applications. [Methods] An L-asparaginase gene was cloned from the metagenome of the fecal microbiota ofNomascus concolor and heterologously expressed inEscherichia coli BL21(DE3). The enzymatic properties of the expressed protein were determined. Furthermore, the potential applications of this protein were explored, including processing potato chips and treating cancer cells. [Results] The cloned L-asparaginase gene,NCasn5, was 996 bp. It encoded the recombinant enzyme NCasn5 with a molecular weight of 37.296 kDa, optimal activity at pH 8.0 and 60 ℃,Km of (3.33±0.21) mmol/L,Vmax of (836.30±13.91) µmol/(min·mg), and a serum half-life of about 69 hin vitro at 37 ℃. NCasn5 reduced the acrylamide content in potato chips by 69.35% and inhibited the growth of human liver cancer cells (QGY-7703) and human melanoma cells (A-375). [Conclusion] We obtained a novel L-asparaginase demonstrating good thermal stability and a prolonged serum half-life. This enzyme lacks the glutaminase activity and reduces acrylamide levels in potato chips. Moreover, it can induce apoptosis in the cancer cell lines QGY-7703 and A-375. These findings suggest the potential applicability of L-asparaginase in both food processing and pharmaceutical industries.
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| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |
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