Brar S K, Verma M, Tyagi R D, Valéro J R, Surampalli R Y
INRS Eau, Terre et Environnement (INRS-ETE), 490, de la Couronne, CP 7500, Québec, Canada G1K 9A9.
Environ Technol. 2008 Feb;29(2):161-70. doi: 10.1080/09593330802028550.
This study investigated the presence of chitinases in Bacillus thuringiensis ssp kurstaki HD-1 (Bt) fermented broths of wastewater sludge (non-hydrolyzed and hydrolyzed); starch industry wastewater and soyameal. Chitinase activity was absent in soyameal and present in others. Chitinase demonstrated peaks at pH 4.0 and temperatures 40 and 50 degrees C with higher activity between pH 4-5 and 10-11. The chitinase band on SDS-PAGE was found to be between 36 and 45 kDa for non-hydrolyzed (NH) and hydrolyzed sludge (TH) and starch industry wastewater. The chitinase profile during fermentation showed peaks at 15 and 30 h for non-hydrolyzed and hydrolyzed sludge and 15 and 24 h for starch industry wastewater. Chitinase retained 96-99 % activity after two weeks incubation at room temperature and pH 4. Bioassays with supplementation of Bt chitinases showed 1.2 fold increase in entomotoxicity of wastewater sludge and a small increase in starch industry wastewater. This study sheds light on production of Bt chitinases in alternative media which will have a long term effect on entomotoxicity of these formulations.
本研究调查了苏云金芽孢杆菌库尔斯塔克亚种HD-1(Bt)在废水污泥(未水解和水解)、淀粉工业废水和豆粕发酵液中几丁质酶的存在情况。豆粕中不存在几丁质酶活性,而其他样品中存在。几丁质酶在pH 4.0以及40和50摄氏度时出现活性峰值,在pH 4 - 5和10 - 11之间活性更高。对于未水解(NH)和水解污泥(TH)以及淀粉工业废水,SDS - PAGE上的几丁质酶条带位于36至45 kDa之间。发酵过程中的几丁质酶谱显示,未水解和水解污泥在15和30小时出现峰值,淀粉工业废水在15和24小时出现峰值。在室温及pH 4条件下孵育两周后,几丁质酶保留了96 - 99%的活性。补充Bt几丁质酶的生物测定表明,废水污泥的昆虫毒性增加了1.2倍,淀粉工业废水的昆虫毒性略有增加。本研究揭示了在替代培养基中生产Bt几丁质酶的情况,这将对这些制剂的昆虫毒性产生长期影响。