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. 2011 Dec;61(4):801-807.
doi: 10.1007/s13213-011-0198-5. Epub 2011 Jan 22.

Purification and characterization of chitinase from Alcaligenes faecalis AU02 by utilizing marine wastes and its antioxidant activity

Neelamegam Annamalai  1 , Mayavan Veeramuthu Rajeswari, Shanmugam Vijayalakshmi, Thangavel Balasubramanian
Affiliations

Purification and characterization of chitinase from Alcaligenes faecalis AU02 by utilizing marine wastes and its antioxidant activity

Neelamegam Annamalai et al. Ann Microbiol. 2011 Dec.

Abstract

Marine waste is an abundant renewable source for the recovery of several value added metabolites with potential industrial applications. This study describes the production of chitinase on marine waste, with the subsequent use of the same marine waste for the extraction of antioxidants. A chitinase-producing bacterium isolated from seafood effluent was identified as Alcaligenes faecalis AU02. Optimal chitinase production was obtained in culture conditions of 37°C for 72 h in 100 ml medium containing 1% shrimp and crab shell powder (1:1) (w/v), 0.1% K(2)HPO(4), and 0.05% MgSO(4)·7H(2)O. The molecular weight of chitinase was determined by SDS-PAGE to be 36 kDa. The optimum pH, temperature, pH stability, and thermal stability of chitinase were about 8, 37°C, 5-12, and 40-80°C, respectively. The antioxidant activity of A. faecalis AU02 culture supernatant was determined through scavenging ability on 1,1-diphenyl-2-picrylhydrazyl (DPPH) as 84%, and the antioxidant compound was characterized by TLC and its FT-IR spectrum. The present study proposed that marine wastes can be utilized to generate a high-value-added product and that pharmacological studies can extend its use to the field of medicine.

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Figures

Fig. 1
Fig. 1
Time course of cell growth and chitinase production of Alcaligenes faecalis AU02 on 1% mixed shrimp/crab shell powder (SCSP) medium. Value represent mean ± SD
Fig. 2
Fig. 2
SDS-PAGE analysis of chitinase from A. faecalis AU02. Lanes: 1 Purified enzyme, 2 molecular markers (29–205 kDa)
Fig. 3
Fig. 3
Effect of temperature on activity and stability of purified chitinase from A. faecalis AU02. Values are mean ± SD, n = 3
Fig. 4
Fig. 4
Effect of pH on activity and stability of purified chitinase from A. faecalis AU02. Values are mean ± SD, n = 3
Fig. 5
Fig. 5
Thin layer chromatography (TLC) analysis of antioxidant materials in the culture supernatant of A. faecalis AU02 grown on 1% SCSP medium. After developing, the TLC plate was visualized by spraying with ninhydrin reagent. Lanes: S N-Standard chitooligosaccharides, A sample
Fig. 6
Fig. 6
Fourier transform-infra red (FT-IR) spectrum of the antioxidant over the wave number range of 4,000–400 cm−1
See this image and copyright information in PMC

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