Food and Fermentation Industries >

2023 , Vol. 49 >Issue 20: 73 - 80

DOI: https://doi.org/10.13995/j.cnki.11-1802/ts.034079

Establishment and optimization of high-throughput screening model for acarbose production by Actinoplanes sp. SE50

  • ZHOU Jian ,
  • LI Xiangfei ,
  • LI Chuang ,
  • LIU Kun ,
  • ZHAO Shiguang ,
  • XUE Zhenglian
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  • 1(College of Biological and Food Engineering, Anhui Polytechnic University, Wuhu 241000, China)
    2(Anhui Engineering Laboratory for Industrial Microbiology Molecular Beeding, Wuhu 241000, China)

Received date: 2022-10-24

  Revised date: 2022-12-12

  Online published: 2023-11-20

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Abstract

Actinoplanes sp. SE50 is the main producing strain of acarbose, a therapeutic drug for type 2 diabetes mellitus. But there are still bottlenecks in the efficient detection of acarbose. In this study, the high-throughput detection model of acarbose was constructed using enzyme detection technology and culture in microwell plate, so as to significantly enhance the screening efficiency of acarbose-producing strains. The results of combined orifice plate fermentation detected by enzyme labeling instrument showed that the correlation between 24-well plate fermentation (R2=0.881 61) and shake flask fermentation was better than that of 48-well plate fermentation (R2=0.833 74). The results of 24-well plate fermentation showed that there was edge effect in 24-well plate fermentation, that was, acarbose yield middle well 1 < long edge well 2 < short edge well 3, which could effectively eliminate the edge effect by filling the edge well with sterile water. Then, to further reduce the process of seed liquid culture, through the orthogonal optimization of the addition amount of soybean cake powder, glycerol and calcium carbonate in the culture medium, the best ratio of 3% soybean cake powder, 1.5% glycerol and 0.4% CaCO3 was obtained. Finally, under the optimal conditions, the yield of acarbose could reach (2 183.66 ±6.60) μg/mL after 168 h of plate fermentation. This study introduces a method of detection and screening of microwell plate culture combined with microplate reader, which greatly shortens the workload and time needed for liquid phase detection of shake flask fermentation, and lays a foundation for screening acarbose-producing strains.

Key words： acarbose; Actinoplanes sp. SE50; high-throughput screening; culture in microwell plate; fermentation optimization

Cite this article

ZHOU Jian , LI Xiangfei , LI Chuang , LIU Kun , ZHAO Shiguang , XUE Zhenglian . Establishment and optimization of high-throughput screening model for acarbose production by Actinoplanes sp. SE50[J]. Food and Fermentation Industries, 2023 , 49(20) : 73 -80 . DOI: 10.13995/j.cnki.11-1802/ts.034079

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