http://www.sciencedirect.com/science?_ob=ArticleURL&_udi=B6TG1-4CJVK1F-1&_user=6763742&_coverDate=08%2F05%2F2004&_rdoc=1&_fmt=high&_orig=search&_sort=d&_doca nchor=&view=c&_searchStrId=1362377336&_rerunOrigin=scholar.google&_acct=C000070526&_versio n=1&_urlVersion=0&_userid=6763742&md5=5a8f934f9c0b027993d3b27a9a5dd034
Characteristics of thermostable chitinase enzymes from the
indonesian
Bacillus
sp.13.26
Purwani. E Yulia, Maggy Thenawidjaja Suhartono, a, Yaya Rukayadib, Jae Kwan Hwangb and Yu Ryang Pyunb
a
Department of Food Technology & Human Nutrition FATETA and Research Center for Biotechnology, Bogor Agricultural University, Darmaga Campus IPB, P.O. BOX 220, Bogor
16002, West Java, Indonesia
b
Bioproduct Research Center, Yonsei University, Seoul, South Korea Received 25 July 2003;
Revised 23 February 2004; accepted 3 March 2004. Available online 7 June 2004.
Abstract
A chitinase producing bacteria was screened and isolated from Tompaso hot spring in North Sulawesi Indonesia. The gram positive spore forming rod shaped bacteria was identified as
Bacillus sp. 13.26 through sequence analysis of the 16SrRNA gene. When grown in medium
containing 0.5% chitin at 55 °C, the thermophillic bacteria optimally produced extracellular chitinase enzyme after 72 h of incubation. The enzyme was purified through ammonium sulfate fractionation, dialysis, heat treatment at 60 °C for 3 h and chromatographed onto DEAE Sepharose CL-6B.
The optimum temperature and pH of the 60-kD chitinase were found at 60 °C and pH 7–8. The thermostable chitinase still retained the activity after incubation for 5 h incubation at 70 °C. Zymogram analysis confirmed the thermostability of the enzyme ; when heated at 80 °C, the enzyme was still significantly active even after 1 h of incubation.
