title: Purification and partial characterization of intact and truncated chitinase from Bacillus thuringiensis HZP7 expressed in Escherichia coli.
abstract: OBJECTIVE
  To ascertain the effect of chitin-binding domain (ChBD) and fibronectin type III domain (FN3) on the characterization of the intact chitinase from Bacillus thuringiensis.
  
  
  RESULTS
  An intact chitinase gene (chi74) from B. thuringiensis HZP7 and its truncated genes (chi54, chi63 and chi66) were expressed in Escherichia coli BL21. The expression products were analyzed after purification. All chitinases were active from pH 4-7.5 and from 20 to 80 °C with identical optimal: pH 5.5 and 60 °C. The activity of colloid chitin degradation for Chi74 was the highest, followed by Chi66, Chi63 and Chi54. Ag(+) reduced the activity of Chi74, Chi54, Chi63 and Chi66, but Mg(2+) enhanced them. The effect of Ag(+) and Mg(2+) was more significant on the activity of Chi54 than on the activities of Chi63, Chi66 and Chi74.
  
  
  CONCLUSIONS
  ChBDChi74 and FN3Chi74 domains play a role in exerting enzymatic activity and can improve the stability of chitinase.
authors.affiliation: Key Laboratory of Biopesticide and Chemical Biology, Ministry of Education, Fujian Agriculture and Forestry University, Fuzhou, 350002, People's Republic of China. shali923@163.com.|School of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou, 350002, People's Republic of China. shali923@163.com.
authors.forename: Li
authors.initials: L
authors.lastname: Sha
authors.affiliation: Key Laboratory of Biopesticide and Chemical Biology, Ministry of Education, Fujian Agriculture and Forestry University, Fuzhou, 350002, People's Republic of China. scientistshao@126.com.|National Engineering Research Center of Juncao Technology, Fujian Agriculture and Forestry University, Fuzhou, 350002, People's Republic of China. scientistshao@126.com.
authors.forename: Ensi
authors.initials: E
authors.lastname: Shao
authors.affiliation: Key Laboratory of Biopesticide and Chemical Biology, Ministry of Education, Fujian Agriculture and Forestry University, Fuzhou, 350002, People's Republic of China. guanxfafu@126.com.
authors.forename: Xiong
authors.initials: X
authors.lastname: Guan
authors.affiliation: Key Laboratory of Biopesticide and Chemical Biology, Ministry of Education, Fujian Agriculture and Forestry University, Fuzhou, 350002, People's Republic of China. zhipengh6841@163.com.|School of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou, 350002, People's Republic of China. zhipengh6841@163.com.
authors.forename: Zhipeng
authors.initials: Z
authors.lastname: Huang
mesh_terms: D001413:Bacillus thuringiensis / Q000201:enzymology*; D002686:Chitin / Q000378:metabolism; D002688:Chitinases / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D004252:DNA Mutational Analysis; D020536:Enzyme Activators / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D015870:Gene Expression; D006863:Hydrogen-Ion Concentration; D007477:Ions / Q000378:metabolism; D008274:Magnesium / Q000378:metabolism; D050505:Mutant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D012834:Silver / Q000378:metabolism; D013696:Temperature
chemical_list: D020536:Enzyme Activators; D007477:Ions; D050505:Mutant Proteins; D011994:Recombinant Proteins; D002686:Chitin; D012834:Silver; D002688:Chitinases; D008274:Magnesium
keywords: Bacillus thuringiensis; Chitin-binding domain; Enzymatic characterization; Fibronectin type III domain; Truncated chitinase