Production and purification of a protease, a chitosanase, and chitin oligosaccharides by Bacillus cereus TKU022 fermentation.
A protease- and chitosanase-producing strain was isolated and identified as Bacillus cereus TKU022. The protease and chitosanase were both produced using 1.5% (w/v) shrimp head powder (SHP) as the sole carbon/nitrogen source, and these enzymes were purified from the culture supernatant. The molecular masses of the TKU022 protease and chitosanase determined using SDS-PAGE were approximately 45 and 44kDa, respectively. The high stability of the TKU022 protease toward surfactants, an optimal pH of 10 and an optimal temperature of 50-60°C suggest that this high-alkaline protease has potential applications for various industrial processes. Concomitant with the production of the TKU022 chitosanase, N-acetyl chitooligosaccharides were also observed in the culture supernatant, including (GlcNAc)(2), (GlcNAc)(4), (GlcNAc)(5), and (GlcNAc)(6) at concentrations of 201.5, 12.4, 0.5, and 0.3μg/mL, respectively, as determined using an HPLC analysis. The chitin oligosaccharides products were also characterized using a MALDI-TOF mass spectrometer. A combination of the HPLC and MALDI-TOF MS results showed that the chitin oligosaccharides of the TKU022 culture supernatant comprise oligomers with degree of polymerization (DP) from 2 to 6. Using this method, the production of a protease, a chitosanase, and chitin oligosaccharides may be useful for various industrial and biological applications.
Life Science Development Center, Tamkang University, New Taipei City, Taiwan.
Tzu-Wen
TW
Liang
Jia-Lin
JL
Hsieh
San-Lang
SL
Wang
D000818:Animals; D001409:Bacillus cereus / Q000737:chemistry / Q000378:metabolism*; D002686:Chitin / Q000096:biosynthesis / Q000302:isolation & purification*; D048271:Chitosan / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D004795:Enzyme Stability; D005285:Fermentation; D006026:Glycoside Hydrolases / Q000096:biosynthesis / Q000302:isolation & purification*; D006863:Hydrogen-Ion Concentration; D008970:Molecular Weight; D009844:Oligosaccharides / Q000096:biosynthesis / Q000302:isolation & purification*; D033561:Penaeidae / Q000737:chemistry; D011208:Powders / Q000737:chemistry; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013379:Substrate Specificity; D013501:Surface-Active Agents; D013696:Temperature; D014866:Waste Products
D009844:Oligosaccharides; D011208:Powders; D013501:Surface-Active Agents; D014866:Waste Products; D002686:Chitin; D048271:Chitosan; D006026:Glycoside Hydrolases; C019868:chitosanase