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High level expression of recombinant horseshoe crab factor C enzymogen and its application in endotoxin detection
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Fu CHEN1, 2, Wenqing WU2, Tingting LIU2, 3, Jiahui ZENG2, 3, Meiqing DENG2, 3, Xiuhua LIN2, 3, Qingping WU2, Ling CHEN2, 3, 4
Acta Microbiologica Sinica | 2026, 66(2) : 899 - 914
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Acta Microbiologica Sinica | 2026, 66(2): 899-914
• Technology and Method •
High level expression of recombinant horseshoe crab factor C enzymogen and its application in endotoxin detection
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Fu CHEN1, 2, Wenqing WU2, Tingting LIU2, 3, Jiahui ZENG2, 3, Meiqing DENG2, 3, Xiuhua LIN2, 3, Qingping WU2, Ling CHEN2, 3, 4
Affiliations
  • 1.School of Biology and Biological Engineering, South China University of Technology, Guangzhou, Guangdong, China
  • 2.State Key Laboratory of Applied Microbiology Southern China, Guangdong Provincial Key Laboratory of Microbial Safety and Health, National Health Commission Science and Technology Innovation Platform for Nutrition and Safety of Microbial Food, Key Laboratory of Big Data Technologies for Food Microbiological Safety (State Administration for Market Regulation), Institute of Microbiology, Guangdong Academy of Sciences, Guangzhou, Guangdong, China
  • 3.Food and Drug Laboratory, Guangdong Detection Center of Microbiology, Guangzhou, Guangdong, China
  • 4.Guangdong Huankai Biological Sci & Tech Co. , Ltd. , Zhaoqing, Guangdong, China
Published: 2026-02-04 doi: 10.13343/j.cnki.wsxb.20250609
Outline
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[Objective] To develop a low-cost and highly sensitive endotoxin detection reagent and detection method with recombinant horseshoe crab factor C enzymogen (rFC). [Methods] The Bac-to-Bac baculovirus expression system was used to express rFC in Sf9 cells and the activity of rFC was measured by the end-point fluorescence assay with endotoxin. The conditions of protein expression were optimized, and ion exchange was used for crude enzyme separation. An endotoxin detection method with rFC based on end-point fluorescence assay was established after the reaction conditions were optimized. Furthermore, the established method was compared with the conventional limulus amebocyte lysate (LAL). [Results] The expression level of rFC was 110.42 mg/L, increasing by 4.75 times. The linear range of endotoxin detection was 0.005-1.000 EU/mL in 1 h, with a good linearity and the limit of detection being 0.005 EU/mL. The applicability rate of this method for actual samples was 92.45%. The consistency of the detection results was 83.67%, and 89.80% of the samples had consistent detection limits with LAL. [Conclusion] This study achieves the efficient expression of rFC and establishes an endotoxin detection method with higher sensitivity than LAL, which has great potential for application.

baculovirus expression system  /  recombinant horseshoe crab factor C enzymogen  /  endotoxin detection
Fu CHEN, Wenqing WU, Tingting LIU, Jiahui ZENG, Meiqing DENG, Xiuhua LIN, Qingping WU, Ling CHEN. High level expression of recombinant horseshoe crab factor C enzymogen and its application in endotoxin detection[J]. Acta Microbiologica Sinica, 2026 , 66 (2) : 899 -914 . DOI: 10.13343/j.cnki.wsxb.20250609
Funding
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  • the National Key Research and Development Program of China(2022YFF1100700)
Appendix
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Year 2026 volume 66 Issue 2
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202
79
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Article Info
doi: 10.13343/j.cnki.wsxb.20250609
  • Receive Date：2025-08-06
  • Online Date：2026-02-05
  • Published：2026-02-04
Article Data
Affiliations
History
  • Received：2025-08-06
  • Accepted：2025-10-13
Funding
the National Key Research and Development Program of China(2022YFF1100700)
Affiliations
    1.School of Biology and Biological Engineering, South China University of Technology, Guangzhou, Guangdong, China
    2.State Key Laboratory of Applied Microbiology Southern China, Guangdong Provincial Key Laboratory of Microbial Safety and Health, National Health Commission Science and Technology Innovation Platform for Nutrition and Safety of Microbial Food, Key Laboratory of Big Data Technologies for Food Microbiological Safety (State Administration for Market Regulation), Institute of Microbiology, Guangdong Academy of Sciences, Guangzhou, Guangdong, China
    3.Food and Drug Laboratory, Guangdong Detection Center of Microbiology, Guangzhou, Guangdong, China
    4.Guangdong Huankai Biological Sci & Tech Co. , Ltd. , Zhaoqing, Guangdong, China

Corresponding:

E-mail: WU Qingping,
CHEN Ling,
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表12种不同金属材料的力学参数
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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