Standardized production of a homogeneous latex enzyme source overcoming seasonality and microenvironmental variables

Prep Biochem Biotechnol. 2021;51(4):375-385. doi: 10.1080/10826068.2020.1818258. Epub 2020 Sep 17.

Abstract

Calotropis procera produces a milky sap containing proteolytic enzymes. At low concentrations, they induce milk-clotting (60 µg/ml) and to dehair hides (0.05 and 0.1%). A protocol for obtaining the enzymes is reported. The latex was mixed with distilled water and the mixture was cleaned through centrifugation. It was dialyzed with distilled water and centrifuged again to recover the soluble fraction [EP]. The dialyze is a key feature of the process. EP was characterized in terms of protein profile, chemical stability, among other criteria. Wild plants belonging to ten geographic regions and grown in different ecological conditions were used as latex source. Collections were carried out, spaced at three-month, according to the seasons at the site of the study. Proteolytic activity was measured as an internal marker and for determining stability of the samples. EP was also analyzed for metal content and microbiology. EP showed similar magnitude of proteolysis, chromatographic and electrophoretic profiles of proteins. Samples stored at 25 °C exhibited reduced solubility (11%) and proteolytic capacity (11%) after six months. Enzyme autolysis was negligible. Microbiological and metal analyses revealed standard quality of all the samples tested. EP induced milk clotting and hide dehairing after storage for up to six months.

Keywords: Bioprocess; Calotropis procera; biotechnology inputs; latex; peptidases.

MeSH terms

  • Animal Fur / drug effects
  • Animals
  • Aspartic Acid Endopeptidases / analysis
  • Aspartic Acid Endopeptidases / chemistry
  • Aspartic Acid Endopeptidases / metabolism*
  • Aspartic Acid Endopeptidases / pharmacology
  • Calotropis / enzymology*
  • Cattle
  • Chemistry Techniques, Analytical / standards*
  • Ecosystem*
  • Goats
  • Hair Removal / methods
  • Latex / chemistry*
  • Latex / isolation & purification
  • Plant Proteins / analysis
  • Plant Proteins / chemistry
  • Plant Proteins / metabolism*
  • Plant Proteins / pharmacology
  • Proteolysis
  • Reference Standards
  • Seasons*
  • Solubility

Substances

  • Latex
  • Plant Proteins
  • Aspartic Acid Endopeptidases
  • rennin-like enzyme (Aspergillus ochraceus)