Characteristics of thermostable chitinase enzymes from the indonesian Bacillus sp.13.26
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.
Author Keywords: Chitinase; Heat stable enzyme
http://www.sciencedirect.com/science?_ob=ArticleURL&_udi=B6TG1-4CJVK1F1&_user=6763742&_coverDate=08%2F05%2F2004&_rdoc=1&_fmt=high&_orig=search&_sort=d&_doca
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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.
Author Keywords: Chitinase; Heat stable enzyme
http://www.sciencedirect.com/science?_ob=ArticleURL&_udi=B6TG1-4CJVK1F1&_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