A β-1,3-endoglucanase produced by Streptomyces rutgersensis was purified to a homogeneity by the fractional precipitation with ammonium sulfate, ion exchange chromatography on Q-Sepharose and hydrophobic chromatography on Butyl Sepharose. A typical procedure provided 11.74-fold purification with 12.53 % yield. SDSPAGE of the purified protein showed one protein band. The exact molecular mass of the enzyme obtained by mass spectrometry was 41.25 kDa; the isoelectric point was between pH 4.2–4.4. The optimal β-glucanase catalytic activity was at pH 7 and 50 °C. An enzyme was only active toward glucose polymers containing β-1,3 linkages and hydrolyzed Saccharomyces cerevisiae cell wall β-glucan in an endo-like way: reaction products were different molecular size β-glucans, which were larger than glucose.
Bacterial strain 68b was isolated from contaminated soil. According to 16S rDNA analysis it belongs to genus Arthrobacter. This strain is capable to utilise phthalic acid as a sole carbon source. This ability was proved by physiological and biochemical tests. By using resting cells, it was found out that Arthrobacter sp. 68b cells could use phthalic acid or convert quinolinic acid if they were pre-grown in the presence of phthalic acid. While analysing the results of a partially sequenced genome, the putative phthalate degradation operon (pht) was detected. It consisted of eight genes; seven genes could code the conversion of phthalate to protocatechuate. It was determined that the gene (pehA) of putative phthalate ester hydrolase is located upstream of pht operon. Genes of putative phthalate degradation operon were re-sequenced and their sequences fully corresponded to the de novo sequencing data. The homology search of genes revealed that all gene products are most similar to phthalate degradation proteins from other Arthrobacter spp. strains and confirmed that the strain 68b converts phthalate to protocatechuate by 3,4-dioxygenase pathway.
β-Glucan is a natural polymer, which is widely studied due to its multiple immunomodulatory properties. In addition, recent findings indicate potent antitumor properties of β-glucan. Saccharomyces cerevisiae, baker's yeast, is one of the commonly used sources of β-1,3-glucan. The aim of this work was to investigate S. cerevisiae β-glucan immunomodulatory activity against cancer cells. In our experiments, BALB/c mice were fed with insoluble whole β-glucan particles, and then their blood was collected for experiments. MH-22a hepatoma cells were treated with the blood of mice fed with β-glucan, and tumor cell viability was investigated after the treatment. The obtained results demonstrated that leukocytes in vivo primed with whole glucan particles, in combination with soluble β-glucan, decreased MH-22a hepatoma cell viability in vitro. Our study has indicated that β-glucan obtained from S. cerevisiae potentially primes mouse whole blood leukocytes to induce cell death of mouse hepatoma cells.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.
hi@scite.ai
10624 S. Eastern Ave., Ste. A-614
Henderson, NV 89052, USA
Copyright © 2024 scite LLC. All rights reserved.
Made with 💙 for researchers
Part of the Research Solutions Family.