Optimization strategy of Bacillus subtilis MT453867 levansucrase and evaluation of levan role in pancreatic cancer treatment

Int J Biol Macromol. 2021 Jul 1:182:1590-1601. doi: 10.1016/j.ijbiomac.2021.05.056. Epub 2021 May 17.

Abstract

Pancreatic cancer is the fourth most lethal cancer type worldwide. Due to multiple levan applications including anticancer activities, studies related to levansucrase production are of interest. To our knowledge, levan effect on pancreatic cancer cells has not been tested previously. In this work, among eighteen bacterial honey isolates, Bacillus subtilis MT453867 showed the highest levan yield (33 g/L) and levansucrase production (8.31 U/mL). One-factor-at-a-time technique increased levansucrase activity by 60% when MgSO4 was eliminated. The addition of 60 g/L banana peels enhanced the enzyme activity (192 U/mL). Placket Burman design determined the media composition for maximum levan yield (54.8 g/L) and levansucrase production (505 U/mL). The identification of levan was confirmed by thin-layer chromatography, Fourier-Transform Infrared spectrometric analysis, 13C-nuclear-magnetic resonance, and 1H-nuclear-magnetic resonance. Both crude and dialyzed levan completely inhibited the pancreatic cancer cell line at 100 ppm with no cytotoxicity on the normal retinal cell line. The LD50 of crude levan was 4833 mg/kg body weight. Levan had strong antioxidant activity and significantly reduced the expression of CXCR4 and MCM7 genes in pancreatic cancer cells with significant DNA fragmentation. In conclusion, Bacillus subtilis MT453867 levan is a promising adjunct to pancreatic-anticancer agents with both anti-cancer and chemoprotective effects.

Keywords: Bacillus subtilis; Honey isolates; Levan; Levansucrase; Pancreatic cancer.

MeSH terms

  • Antineoplastic Agents / metabolism*
  • Antineoplastic Agents / pharmacology
  • Bacillus subtilis / enzymology*
  • Bacillus subtilis / metabolism*
  • DNA Fragmentation / drug effects
  • Fructans / metabolism*
  • Fructans / pharmacology
  • Hexosyltransferases / metabolism*
  • Humans
  • Minichromosome Maintenance Complex Component 7 / metabolism
  • Pancreatic Neoplasms / metabolism
  • Receptors, CXCR4 / metabolism
  • Spectroscopy, Fourier Transform Infrared

Substances

  • Antineoplastic Agents
  • CXCR4 protein, human
  • Fructans
  • Receptors, CXCR4
  • levan
  • Hexosyltransferases
  • levansucrase
  • MCM7 protein, human
  • Minichromosome Maintenance Complex Component 7