Abstract
A strain isolated from the feces of takin was identified asClostridium aminovalericum. In response to various types of chitin used as growth substrates, the bacterium produced a complete array of chitinolytic enzymes: chitinase (‘endochitinase’), exochitinase, β-N-acetylglucosaminidase, chitosanase and chitin deacetylase. The highest activities of chitinase (536 pkat/mL) and exochitinase (747 pkat/mL) were induced by colloidal chitin. Fungal chitin also induced high levels of these enzymes (463 pkat/mL and 502 pkat/mL, respectively). Crab shell chitin was the best inducer of chitosanase activity (232 pkat/mL). The chitinolytic enzymes of this strain were separated from culture filtrate by ion-exchange chromatography on the carboxylic sorbent Polygran 27. At pH 4.5, some isoforms of the chitinolytic enzymes (30 % of total enzyme activity) did not bind to Polygran 27. The enzymes were eluted under a stepwise pH gradient (pH 5–8) in 0.1 mol/L phosphate buffer. At merely acidic pH (4.5–5.5), the adsorbed enzymes were co-eluted. However, at pH close to neutral values, the peaks of highly purified isoforms of exochitinases and chitinases were isolated. The protein and enzyme recovery reached 90 %.
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Šimůnek, J., Tishchenko, G., Rozhetsky, K. et al. Chitinolytic enzymes fromClostridium aminovalericum: Activity screening and purification. Folia Microbiol 49, 194–198 (2004). https://doi.org/10.1007/BF02931401
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DOI: https://doi.org/10.1007/BF02931401