Indonesian Journal of Biotechnology
Vol 28, No 1 (2023)

Production, purification and characterization of chitinase from Micromonospora sp. AR17

Yohanes Harvinda (Fisheries Science Postgraduate Program, Department of Fisheries, Faculty of Agriculture, Universitas Gadjah Mada, Jalan Flora A4, Bulaksumur, Yogyakarta 55281, Indonesia)
Ustadi Ustadi (Fisheries Science Postgraduate Program, Department of Fisheries, Faculty of Agriculture, Universitas Gadjah Mada, Jalan Flora A4, Bulaksumur, Yogyakarta 55281, Indonesia)
Masagus Muhammad Prima Putra (Fisheries Science Postgraduate Program, Department of Fisheries, Faculty of Agriculture, Universitas Gadjah Mada, Jalan Flora A4, Bulaksumur, Yogyakarta 55281, Indonesia)



Article Info

Publish Date
31 Mar 2023

Abstract

N‐acetylglucosamine (NAG) is the monomer product of chitin, which has been widely used as a bioactive com‐ pound in applications such as anti‐tumor, anti‐microbial, and antioxidant activities. In production, biological processes using enzymes are preferable to chemicals due to environmental issues. This study aims to determine the activity, purity level, and molecular weight of purified chitinase from Micromonospora sp. AR17 determines the concentration of NAG produced by purified chitinase that has been characterized. Chitinase was produced by fermentation in colloidal chitin broth at 40 °C, pH 7, for 7 days, while chitinase activity was checked every 24 h. The optimal fermentation time was used to produce chitinase for a further purification step. Enzyme purification was carried out by ultrafiltration, ammonium sulfate precipitation, ion exchange chromatography (Q Sepharose Fast Flow), and gel filtration (Sephacryl S‐300). The purified enzyme was then char‐ acterized for optimum time, pH, and temperature to produce NAG. The results suggested that the fourth day was the optimal time for chitinase production, with chitinase activity of 0.0040 U/mL and a NAG concentration of 7.62 µg/mL. The purifica‐ tion step successfully increased the purity by 6.82 times with chitinase‐specific activity at 1.4648 U/mg. Production of NAG with purified chitinase produced a NAG concentration of 32.472 µg/mL with an incubation time of 30 min at 40 °C and pH 7.

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Journal Info

Abbrev

ijbiotech

Publisher

Subject

Biochemistry, Genetics & Molecular Biology Immunology & microbiology Materials Science & Nanotechnology

Description

The Indonesian Journal of Biotechnology (IJBiotech) is an open access, peer-reviewed, multidisciplinary journal dedicated to the publication of novel research in all aspects of biotechnology, with particular attention paid to the exploration and development of natural products derived from ...