Thesis

Optimization of Agarase Enzyme Activity from Marine Streptomyces BLH 3-7 with Response Surface Methodology

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Marine Streptomyces BLH 3-7 had been isolated from Bitung Sea, Sulawesi, as bacteria capable of hydrolyzing agar. To optimize the agarase activity from the bacteria, selection of the best temperature for culture condition and modifying the carbon, nitrogen, and mineral range of concentration in the media (e.g., maltose, (NH4 Methodology (RSM) was conducted. The temperature of 37 activity as the temperature of 45o)2SO4, & CaCl2o.2H2O) through Response Surface C was the best temperature for agarase C and above increased the formation of existing reducing sugar in the media, which is detected in the reducing assay (i.e., DNS or 3,5-Dinitrosalicylic Acid assay). The RSM result showed that the usage of 0.5-1.5% of maltose introduced a lot of contaminants as reducing sugar detected in the DNS assay, resulting in a linear relationship between the enzyme activity and
carbon sources alone. Thus, the final enzyme activity measured was compromised. TLC results also
showed that the crude enzyme sample contained too many compounds to be separated on the plate
. No changes in the media compositions were observed over 168 hours or seven days of culture
incubation. Therefore, partial purification, such as ammonium sulfate precipitation, is necessary for
experiments using reducing sugar assay on samples containing carbohydrates with reducing end. To
tackle this problem, this study developed a control enzyme to measure the existing reducing sugar
present in the sample before measuring the actual enzyme activity, however, it is incomplete and
needs to be further designed.

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Availability

Detail Information

Series Title

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Call Number

BT 23-014

Publisher

i3L, Jakarta : i3L, Jakarta., 2023

Collation

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Language

English

ISBN/ISSN

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Classification

NONE

Content Type

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Media Type

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Carrier Type

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Edition

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Subject(s)

Streptomyces BLH 3-7
marine bacteria
Bitung Sea Sulawesi

Specific Detail Info

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Statement of Responsibility

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