Purification and characterization of protease and chitinase from Bacillus cereus TKU006 and conversion of marine wastes by these enzymes.
A chitinase- and protease-producing bacterium was isolated and identified as Bacillus cereus TKU006. The better condition on our tests for protease and chitinase production was found when the culture was shaken at 25 degrees C for 2 days in 25 mL of medium containing 2% shrimp shell powder (w/v), 0.1% K(2)HPO(4), and 0.05% MgSO(4).7H(2)O. The molecular masses of TKU006 protease and chitinase determined by sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis were approximately 39 and 35 kDa, respectively. The optimum pH, optimum temperature, pH stability, and thermal stability of TKU006 protease and chitinase were 9, 50 degrees C, 3-11, 50 degrees C and 5, 40 degrees C, 3-11, 60 degrees C, respectively. TKU006 protease was inhibited completely by EDTA, indicating that the TKU006 protease was a metalloprotease. The TKU006 protease and chitinase retained 61%, 60%, 73%, and 100% and 60%, 60%, 71%, and 96% of its original activity in the presence of 2% Tween 20, 2% Tween 40, 2% Triton X-100, and 1 mM SDS, respectively. The antioxidant activity of TKU006 culture supernatant was determined through the scavenging ability on DPPH with 70% per milliliter. In conclusion, the novelties of the TKU006 protease and chitinase include its high stability to the surfactants and pH. Besides, with this method, we have shown that marine wastes can be utilized to generate a high-value-added product and have revealed its hidden potential in the production of functional foods.
Wang
San-Lang
SL
Graduate Institute of Life Sciences, Tamkang University, Tamsui, Taiwan. sabulo@mail.tku.edu.tw
Chao
Chia-Hsing
CH
Liang
Tzu-Wen
TW
Chen
Chung-Chih
CC
D001409:Bacillus cereus / Q000201:enzymology*; D001713:Biphenyl Compounds; D002688:Chitinases / Q000235:genetics* / Q000378:metabolism; D002855:Chromatography, Thin Layer; D004492:Edetic Acid; D004591:Electrophoresis, Polyacrylamide Gel; D006863:Hydrogen-Ion Concentration; D010447:Peptide Hydrolases / Q000235:genetics* / Q000378:metabolism; D010851:Picrates; D011136:Polysorbates; D013696:Temperature
D001713:Biphenyl Compounds; D010851:Picrates; D011136:Polysorbates; D004492:Edetic Acid; C004931:1,1-diphenyl-2-picrylhydrazyl; D002688:Chitinases; D010447:Peptide Hydrolases