Single-step purification of chitosanases from Bacillus cereus using expanded bed chromatography

Abstract

A chitosanase-producing strain was isolated and identified as Bacillus cereus C-01. The purification and characterization of two chitosanases were studied. The purification assay was accomplished by ion exchange expanded-bed chromatography. Experiments were carried out in the presence and in the absence of cells through different expansion degree to evaluate the process performance. The adsorption experiments demonstrated that the biomass does not affect substantially the adsorption capacity of the matrix. The enzyme bound to the resin with the same extent using clarified and unclarified broth (0.32 and 0.30 U/g adsorbent, respectively). The fraction recovered exhibited 31% of the yield with a 1.26-fold increase on the specific activity concerned to the initial broth. Two chitosanases from different elution steps were recovery. Chit A and Chit B were stable at 30-60°C, pH 5.5-8.0 and 5.5-7.5, respectively. The highest activity was found at 55°C, pH 5.5 to Chit A and 50°C, pH 6.5 to Chit B. The ions Cu(2+), Fe(2+) and Zn(2+) indicated inhibitory effect on chitosanases activities that were significantly activated by Mn(2+). The methodology applied in this study enables the partial purification of a stable chitosanase using a feedstock without any pre-treatment using a single-step purification.

Purification and Characterization of A New Cold-Adapted and Thermo-Tolerant Chitosanase from Marine Bacterium Pseudoalteromonas sp. SY39

“…It is produced by the deacetylation of chitin which is the second-most ubiquitous polysaccharide in nature [1,2,3]. Chitosan polymers are widely used as absorbable surgical suture, artificial skin and wound healing accelerators mainly due to their high viscosity and nontoxicity [4,5,6].…”

Optimization of chitosanase production by Bacillus mojavensis EGE‐B‐5.2i

“…Work on purification and characterization of chitosanase is still in progress. Bacillus cereus CH12 11.16 [27] B. cereus C-01 0.8 [31] Bacillus mycoides TKU038 0.58 [32] Bacillus thuringiensis ZJOU-010 4.25 [21] Bacillus cereus TKU018 22 [33] Bacillus sp. RKY3.…”

Bioprocessing of Squid Pens Waste into Chitosanase by Paenibacillus sp. TKU047 and Its Application in Low-Molecular Weight Chitosan Oligosaccharides Production

“…However, the biggest obstacle to adopting this method is the expensive cost of the enzyme. To reduce the cost of the enzyme and that of COS produced by the enzymatic method, some solutions include reducing the enzyme purification steps [ 18 ], using a commercial enzyme complex to hydrolyze chitosan [ 19 ], immobilizing the enzyme [ 20 ], using ultrafiltration membrane enzymatic system [ 21 ], and utilizing low-cost materials to produce chitinolytic enzymes via microbial fermentation [ 22 ].…”

“…Endo-chitosanase degrades chitosan at a random position and releases COS, thus it is the major enzyme responsible for COS production [ 2 ]. Until now, numerous bacteria have been explored for their chitosanase producing ability, such as Bacillus [ 10 , 15 , 18 , 23 , 24 , 25 , 26 , 27 , 28 , 29 ], Paenibacillus [ 8 , 13 , 30 , 31 , 32 , 33 , 34 , 35 , 36 , 37 , 38 , 39 , 40 , 41 ], Acinetobacter [ 42 , 43 ], Streptomyces [ 14 , 44 ], Serratia [ 45 ], and Pseudomonas [ 46 ]. For the production of chitosanase via bacterial fermentation, chitin is the common carbon and nitrogen (C/N) supplement.…”