Functional characterization of recombinant bromelain
Stem bromelain is a plant protease with a number of industrial and therapeutic applications. This study investigated the functional properties of commercial bromelain and recombinant bromelain (expressed in E. coli BL 21-AI). In vitro cytotoxicity assays were conducted to evaluate the effect of b...
| Main Authors: | , , |
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| Format: | Conference or Workshop Item |
| Language: | English |
| Published: |
2015
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| Subjects: | |
| Online Access: | http://irep.iium.edu.my/45893/ http://irep.iium.edu.my/45893/1/oral_presentation-Azura.pdf |
| Summary: | Stem bromelain is a plant protease with a number of industrial and therapeutic
applications. This study investigated the functional properties of commercial bromelain
and recombinant bromelain (expressed in E. coli BL 21-AI). In vitro cytotoxicity assays
were conducted to evaluate the effect of both bromelains on tumour cell lines (murine
melanoma and breast cancer MCF-7) and normal cells (Vero and Chinese hamster
ovary) using Sulforhodamine B and MTT tests. Anti-inflammatory activities of the
enzymes were also assessed on murine macrophage cell lines using Griess reagents and
ELISA test. The beef tenderizing effects of both bromelains and in-situ localization of the
recombinant bromelain were analysed using Transmission Electron Microscopy. The
results obtained revealed that both bromelains were more cytotoxic to tumour cell lines
(IC50 values ≈ 0.16-0.23mg/mL) than normal cell lines (IC50 values ≈ 0.27-0.51mg/mL).
Moreover, the two enzymes had effectively inhibited the pro-inflammatory mediators’
production and thus, had good anti-inflammatory activity. Both enzymes were found to
have comparative anti-cancer and anti-inflammatory properties. The expression of
recombinant enzyme was discovered to occur in the cytoplasm of E. coli BL 21-AI and
the purified enzyme efficiently degraded both actin and myosin of fresh beef. Hence, it
served as a good meat tenderizer. |
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