Increased Production of Carrageenase by <i>Pseudomonas aeruginosa</i> ZSL-2 Using Taguchi Experimental Design

Abstract

The culture conditions for the production of carrageenase were optimized using one-factor-at-atime method combined with orthogonal array design. With the one-factor-at-a-time method revealed optimal conditions for carrageenase production were 24 h of fermentation period, 28 °C incubation temperature at pH 8.0 with NaNO 3 as nitrogen source and carrageenan as carbon source in MMS media. Further optimization of carrgeenase production by using orthogonal experimental design L 9 (3 4 ) with four factors, temperature, pH, NH 4 NO 3 and carrageenan with their relevant levels revealed optimised conditions for carrageenase production were temperature of 28 °C, pH 8.0, 2 g L -1 NaNO 3 and 2 g L -1 carrageenan. The order of the factors affecting the fermentation process was found to be temperature > pH > NaNO 3 > carrageenan. The temperature played a significant role on the carrageenase production. Higher carrageenase yield with activity of 0.542 ±0.045 U ml -1 was obtained in the optimised medium when compared to those of basal medium. Carrageenase hydrolysed products of carrageenan were identified by LC-ESI-MS as neocarrabiose, neocarrabiose-4 sulfate, neocarratetraose, neocarratetraose-4 sulfate, anhydrogalactose, galactose, galactose-4 sulphate and sulphate.

Optimization of Agrase Production by Alkaline <i>Pseudomonas aeruginosa</i> ZSL-2 Using Taguchi Experimental Design

“…In L 9 orthogonal array method only 9 experiments were performed whereas, if factorial experimental design International Letters of Natural Sciences Vol. 17(Planket Burner design and central composite design) was used, at least 81 experiments in triplicate would be necessary to achieve the same results. Very few reports have described the optimization of agarase production using one factor at time method [10,12,14] reported optimisation of agarase production by statistical strategy using combination of Planket Burner design and central composite design (CCD).…”

“…This strain was routinely maintained and stored at 4 °C on MMS-agar slants in the laboratory. Seed culture was prepared from 24h grown culture on MMS liquid broth at 180 rpm and 37 °C as reported earlier [17].…”

Bacterial carrageenases: an overview of production and biotechnological applications

“…In this regard, studies on important carrageenan degraders reported in the recent years are listed in Table 2. Among bacteria, degradation is mostly confined to Gram negative, mainly various Pseudoalteromonas, Cellulophaga, Pseudomonas, Cytophaga, Tamlana, Vibrio, Catenovulum, Microbulbifer, Zobellia, Alteromonas (Shangyong et al 2013; Yao et al 2013; Ziayoddin et al 2014; Mou et al 2004; Feixue et al 2010; Li et al 2015; Hatada et al 2011; Liu et al 2013; Michel et al 2001b; Zhu and Ning 2016). However, some Gram positive bacteria like Bacillus sp.…”

“…QY3Iota-carrageenan/25 °C/150 rpm/20 h/pH 7.050>80 %/50 °C/pH 7.0/60 min7.0>70 %/pH 5.0–10.6/720 min/4 °C48.3 kDaMa et al (2013)11 Cytophaga lk-C783ZM g having carrageenan/25 °C/150 rpm/72 h25 °C50 %/50 °C/pH 7.0/10 min7.6NR n 100 kDaSarwar et al (1983a, b, 1987)12 Cytophaga MCA-2ZM g having carrageenan/32 °C/150 rpm/36 h/pH 7.5280 %/55 °C/30 min7.232 %/pH 10.83/24 h30 kDaMou et al (2004)13Dsij strainZM g /22 °C/250 rpm/24–40 h40100 %/30 °C/pH 7.2/120 min7.2100 %/7.0/60 min/40 °C40 kDaPotin et al (1991)14Marine bacteriaCarrageenan/20 °C–25 °C/100 rpm/72–96 h40NR n 7.5NR n NR n Yaphe and Baxter (1955)15 Pseudoalteromonas sp. QY203Kappa-carrageenan/25 °C/150 rpm/48 h4570 %/35 °C/pH 7.2/48 h7.2>70 %/6.5–9.0/6 h/4 °C34Shangyong et al (2013)16 Pseudomonas aeruginosa ZSL-2Kappa-carrageenan + MMSM h /37 °C/180 rpm/24 h28NR n 8.0NR n NR n Ziayoddin et al (2014)17 Pseudoalteromonas strain CL19Kappa-carrageenan/20 °C/72 h…”

“…Although the growth of bacteria was increased in the medium co-supplemented with glucose, fructose or maltose. This may be due to rapid utilization of these simple sugars by the bacterium resulting in an increase in cell mass by vigorous growth (Ziayoddin et al 2014; George et al 2014a, b; Sondhi et al 2015). …”