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This experiment investigated the ability of an aminopeptidase gene dmpA from microcyst basidiomycete MCA 4210 (Cystobasidium minutum MCA 4210) to catalyze the synthesis of L-carnosine, and constructed a genetically engineered strain to catalyze the synthesis of L-carnosine with L-histidine and β-alanine methyl ester hydrochloride as substrates, aiming to improve the synthesis efficiency of L-carnosine. Codon-optimized DmpA was expressed in Escherichia coli BL21 (DE3) and adding a maltose binding protein (MBP) tag further improved solubility and enzyme activity, yielding BL21 (DE3)-MBP-DmpA. Then the recombinant strain BL21(DE3)-MBP-DmpA was further investigated to optimize induced expression and transformation conditions. The results indicated that the target gene fragment was successfully fused with MBP, yielding the recombinant strain BL21(DE3)-MBP-DmpA. After optimization of the induction conditions, the crude enzyme activity reached 145 U. Through optimization of the transformation conditions, the L-carnosine production reached as high as 55.72 g/L, with a yield of 54.54%. This study successfully expressed DmpA from Cystobasidium minutum MCA 4210 in E. coli, demonstrating its L-carnosine synthesis capability. The MBP tag and condition optimizations significantly improved production, achieving the highest L-carnosine yield reported to date. These findings provided a strong foundation for industrial-scale L-carnosine production using genetically engineered bacteria and highlighted promising application potential.
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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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