Comparison of Statistical Methods for Optimization of Salts in Medium for Production of Carboxymethylcellulase of Bacillus amyloliquefaciens DL-3 by a Recombinant E. coli JM109/DL-3

Abstract

The optimal concentrations of salts in medium for cell growth and the production of carboxymethylcellulase (CMCase) by a recombinant E. coli JM109/DL-3 were established using two statistical methods: orthogonal array method (OAM) and response surface method (RSM). The analysis of variance (ANOVA) of data based on OAM indicated that K2HPO4 gave maximum sum of square (S) and percentage contribution (P) for cell growth as well as production of CMCase. The optimal concentrations of K2HPO4, NaCl, MgSO4•7H2O, and (NH4)2SO4 in medium for cell growth extracted by Qualitek-4 (W32b) Software were 10.0, 1.0, 0.2, and 0.6 g/l, respectively, whereas those for the production of CMCase by E. coli JM109/DL-3 were 5.0, 1.0, 0.4, and 0.6 g/l. The analysis of variance (ANOVA) resulting from RSM indicated that a highly significant salt for cell growth was K2HPO4 ("probe>F" less than 0.0001), whereas K2HPO4 and MgSO4•7H2O were significant for the production of CMCase. The optimal concentrations of K2HPO4, NaCl, MgSO4•7H2O, and (NH4)2SO4 for cell growth extracted by Design Expert Software were 7.44, 1.08, 0.22, and 0.88 g/l, respectively, whereas those for production of CMCase were 5.84, 0.69, 0.28, and 0.54 g/l. The optimal concentrations of salts and their influences on cell growth and production of CMCase extracted by OAM were almost the same as those by RSM. Production of CMCase by a recombinant E. coli JM109/DL-3 under optimized concentration of salts was 1.93 times higher than that by Bacillus amyloliquifaciens DL-3.

Enhanced Production of Carboxymethylcellulase by a Newly Isolated Marine Microorganism Bacillus atrophaeus LBH-18 Using Rice Bran, a Byproduct from the Rice Processing Industry

“…The ANOVA indicated that this model and the model term of X1 ("probe > F" less 0.0001) were highly significant [23].…”

Construction of Recombinant Escherichia coli JM109/LBH-10 and Comparison of Its Optimal Condition for Production of Carboxymethyl cellulase with Its Wild Type, Psychrobacter aquimaris LBH--10

“…The best carbon and nitrogen sources for cell growth of B. amyloliquefaciens DL-3 were sucrose and malt extract, whereas those for production of CMCase were rice hulls and peptone [22]. However, the best carbon and nitrogen sources for cell growth of its recombinant E. coli JM109/ DL-3 were fructose and ammonium chloride, whereas those for production of CMCase were rice bran and tryptone [23]. The best carbon and nitrogen source for cell growth and production of CMCase by B. subtilis subsp.…”

“…Those for production of CMCases by bacterial and fungal stains ranged from 25 to 37°C, except for thermophilic microorganisms such as Thermoascus aurantiacu, which optimal temperature for production of CMCase is 50°C [23]. The optimal temperature for the cell growth of a psychrophilic marine bacterium, P. aquimaris LBH-10 was lower than that for production of CMCase.…”

Enhanced Production of Cellobiase by a Marine Bacterium, Cellulophaga lytica LBH-14, in Pilot-Scaled Bioreactor Using Rice Bran

“…Potassium phosphate is one of major salts in the medium for productions of microbial polysaccharides and enzymes as well as a well-known ingredient in buffer solutions [13,23]. Sodium chloride was reported to be used as a physiological modulator of biosynthetic pathway of biopolymers [12,24]. Magnesium sulfate added to media assists with spore germination and initial growth of A. fisheri, which results in 1.9 fold increased production of xylanase [31].…”

“…The optimization of culture medium by the traditional "one-factor-at-a-time" method requires a considerable amount of work and time. An alternative strategy is a statistical approach, for example, RSM involving a mini-mum number of experiments for a large number of factors [24].…”