OPTIMIZATION OF CULTURE PARAMETERS FOR α-GLUCOSIDASE PRODUCTION FROM PROTONEMAL BIOMASS OF BRYUM CORONATUM SCHWAEGR
Keywords:
Bryum coronatum, Protonemal biomass, NilAbstract
Objective: The purpose of the present investigation was to optimization of culture parameters for α-glucosidase production from protonemal biomass of Bryum coronatum Schwaegr.
Methods: Fresh unopened, mature capsules were used as explant and the protonema that developed from the aseptic spores were cultured grown on 1/4th Murashige and Skoog basal medium. Frozen protonemal biomass was homogenized in 100 mM phosphate buffer (pH 7.0). The supernatant was assayed for α-glucosidase activity. Various culture parameters such as incubation period, temperature, pH, agitation, carbon sources and nitrogen sources were evaluated and further Taguchi orthogonal array method was performed.
Results: It was observed that both protonemal biomass and α-glucosidase production were maximum at 28 d of culture. Optimization of culture parameters such as pH, temperature and agitation speed for α-glucosidase production was found to be 6.0, 35 °C and 150 rpm, respectively. Among nutritional parameters, sucrose as carbon source and ammonium nitrate as nitrogen source found to be effective for enzyme production and maximum growth of protonemal biomass of B. coronatum. Based on Taguchi orthogonal array method, the optimal condition and their contribution on α-glucosidase production were evaluated as follows: sucrose (1.5%), ammonium nitrate (1%), pH (6.0), temperature (35 °C) and agitation (150 rpm).
Conclusion: In this study, the culture of moss, B. coronatum proved to be a good source for the enzyme α-glucosidase production. Temperature 35 °C and pH 6.0 were found to be optimum for maximum α-glucosidase production with respect to protonemal biomass production. Optimization of culture medium by Taguchi method has resulted in an increase in the α-glucosidase activity from 3.84-14.39 U/ml.
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