Generation and characterization of caprine chymosin in corn seed

Protein Expr Purif. 2017 Jul:135:78-82. doi: 10.1016/j.pep.2017.05.004. Epub 2017 May 17.

Abstract

Chymosin is widely used in the dairy industry, and much is produced through recombinant DNA in organisms such as bacteria and tobacco. In this study, we used a new transgenic method to express caprine chymosin in corn seeds with lower cost and better storage capability. The recombinant chymosin protein was successfully expressed at an average level of 0.37 mg/g dry weight, which is 0.27% of the total soluble protein in the corn seed. Prochymosin can be activated to produce a chymosin protein with the ability to induce clotting in milk, similar to the commercial protein. The activity of the purified recombinant chymosin was as high as 178.5 U/mg. These results indicate that we have successfully established a technology for generating corn seed-derived caprine chymosin for potential use in the dairy industry.

Keywords: Caprine chymosin; Corn seed; Transgenic plants.

MeSH terms

  • Agrobacterium tumefaciens / genetics
  • Agrobacterium tumefaciens / metabolism
  • Animals
  • Chymosin / biosynthesis*
  • Chymosin / genetics
  • Chymosin / isolation & purification
  • Chymosin / pharmacology
  • Cloning, Molecular
  • Enzyme Assays
  • Flocculation / drug effects
  • Food Technology
  • Gene Expression
  • Genetic Vectors / chemistry*
  • Genetic Vectors / metabolism
  • Globulins / genetics
  • Globulins / metabolism
  • Goats
  • Kinetics
  • Milk / chemistry
  • Milk / drug effects
  • Plant Proteins / genetics
  • Plant Proteins / metabolism
  • Plants, Genetically Modified*
  • Promoter Regions, Genetic
  • Recombinant Proteins / biosynthesis
  • Recombinant Proteins / genetics
  • Recombinant Proteins / isolation & purification
  • Recombinant Proteins / pharmacology
  • Seeds / enzymology
  • Seeds / genetics*
  • Transformation, Genetic
  • Zea mays / enzymology
  • Zea mays / genetics*

Substances

  • Globulins
  • Plant Proteins
  • Recombinant Proteins
  • Chymosin