• Journal of Microbiology and Biotechnology
• /
• 제29권7호
• /
• pp.1061-1070
• /
• 2019

In the present study, the optimization of poly(${\gamma}$-glutamic acid) (${\gamma}$-PGA) production by Bacillus sp. FBL-2 was studied using a statistical approach. One-factor-at-a-time method was used to investigate the effect of carbon sources and nitrogen sources on ${\gamma}$-PGA production and was utilized to select the most significant nutrients affecting the yield of ${\gamma}$-PGA. After identifying effective nutrients, response surface methodology with central composite design (CCD) was used to obtain a mathematical model to identify the optimum concentrations of the key nutrients (sucrose, $\text\tiny{L}$-glutamic acid, yeast extract, and citric acid) for improvement of ${\gamma}$-PGA production. The optimum amount of significant medium components appeared to be sucrose 51.73 g/l, $\text\tiny{L}$-glutamic acid 105.30 g/l, yeast extract 13.25 g/l, and citric acid 10.04 g/l. The optimized medium was validated experimentally, and ${\gamma}$-PGA production increased significantly from 3.59 g/l (0.33 g/l/h) to 44.04 g/l (3.67 g/l/h) when strain FBL-2 was cultivated under the optimal medium developed by the statistical approach, as compared to non-optimized medium.

• Journal of Microbiology and Biotechnology
• /
• 제31권5호
• /
• pp.717-725
• /
• 2021

This study aimed to optimize medium composition and culture conditions for enhancing the biomass of Lactobacillus plantarum 200655 using statistical methods. The one-factor-at-a-time (OFAT) method was used to screen the six carbon sources (glucose, sucrose, maltose, fructose, lactose, and galactose) and six nitrogen sources (peptone, tryptone, soytone, yeast extract, beef extract, and malt extract). Based on the OFAT results, six factors were selected for the Plackett-Burman design (PBD) to evaluate whether the variables had significant effects on the biomass. Maltose, yeast extract, and soytone were assessed as critical factors and therefore applied to response surface methodology (RSM). The optimal medium composition by RSM was composed of 31.29 g/l maltose, 30.27 g/l yeast extract, 39.43 g/l soytone, 5 g/l sodium acetate, 2 g/l K₂HPO₄, 1 g/l Tween 80, 0.1 g/l MgSO₄·7H₂O, and 0.05 g/l MnSO₄·H₂O, and the maximum biomass was predicted to be 3.951 g/l. Under the optimized medium, the biomass of L. plantarum 200655 was 3.845 g/l, which was similar to the predicted value and 1.58-fold higher than that of the unoptimized medium (2.429 g/l). Furthermore, the biomass increased to 4.505 g/l under optimized cultivation conditions. For lab-scale bioreactor validation, batch fermentation was conducted with a 5-L bioreactor containing 3.5 L of optimized medium. As a result, the highest yield of biomass (5.866 g/l) was obtained after 18 h of incubation at 30℃, pH 6.5, and 200 rpm. In conclusion, mass production by L. plantarum 200655 could be enhanced to obtain higher yields than that in MRS medium

• Journal of Microbiology and Biotechnology
• /
• 제18권1호
• /
• pp.138-144
• /
• 2008

The production of the entomopathogenic and phytotoxic cyclic depsipeptide beauvericin (BEA) was studied in submerged cultures of Fusarium oxysporum KFCC 11363P isolated in Korea. The influences of various factors on mycelia growth and BEA production were examined in both complete and chemically defined culture media. The mycelia growth and BEA production were highest in Fusarium defined medium. The optimal carbon and nitrogen sources for maximizing BEA production were glucose and $NaNO_3$, respectively. The carbon/nitrogen ratio for maximal production of BEA was investigated using response surface methodology (RSM). Equations derived by differentiation of the RSM model revealed that the production of BEA was maximal when using 108 mM glucose and 25 mM $NaNO_3$.

• 한국축산식품학회지
• /
• 제30권1호
• /
• pp.66-72
• /
• 2010

Pediococcus pentosaceus SA131 was isolated from jeotgal, is the bacteriocin producer against bovine mastitis pathogens, Streptococcus uberis E290, Enterococcus gallinarum E362, and Staphylococcus epidermidis ATCC 12228. The medium composition for pediocin SA131 production by P. pentosaceus SA131 was optimized using response surface methodology. Component of medium was studied as carbon source (glucose, fructose, lactose, glycerol, sucrose, maltose, and mannitol), nitrogen source (beef extract, yeast extract, peptone, malt extract, and tryptone), mineral and surfactant ($MgSO_4$, $KH_2PO_4$, $(NH_4)_2SO_4$, $MnSO_4$, NaCl, sodium acetate, and Tween 80). Through one factor-at-a-time experiment, glucose, fructose, yeast extract, malt extract, NaCl, $MgSO_4$, and Tween 80 were determined as the good ingredient. The effects of major factors for pediocin SA131 production were investigated by two-level fractional factorial designs (FFD). By a $2^4$ FFD, fructose, yeast extract, and $MnSO_4$ were found to be the important factors for the bacteriocin production. Subsequently, a $2^3$ central composite design (CCD) was adopted to derive a statistical model for optimizing the composition of the fermentation medium. The estimated optimum composition for the production of pediocin SA131 by P. pentosaceus SA131 was as follows; 0.13% fructose, 1% glucose, 1.8% yeast extract, 2.58% $MnSO_4$, 0.2% NaCl, and 0.2% Tween 80. The pediocin production under optimized medium was increased to 1,000 AU/mL, compared to the 400 AU/mL in MRS medium.

• Journal of Microbiology and Biotechnology
• /
• 제23권12호
• /
• pp.1757-1764
• /
• 2013

Trichoderma harzianum ZF-2 producing laccase was isolated from decaying samples from Shandong, China, and showed dye decolorization activities. The objective of this study was to optimize its culture conditions using a statistical analysis of its laccase production. The interactions between different fermentation parameters for laccase production were characterized using a Plackett-Burman design and the response surface methodology. The different media components were initially optimized using the conventional one-factor-at-a-time method and an orthogonal test design, and a Plackett-Burman experiment was then performed to evaluate the effects on laccase production. Wheat straw powder, soybean meal, and $CuSO_4$ were all found to have a significant influence on laccase production, and the optimal concentrations of these three factors were then sequentially investigated using the response surface methodology with a central composite design. The resulting optimal medium components for laccase production were determined as follows: wheat straw powder 7.63 g/l, soybean meal 23.07 g/l, $(NH_4)_2SO_4$ 1 g/l, $CuSO_4$ 0.51 g/l, Tween-20 1 g/l, $MgSO_4$ 1 g/l, and $KH_2PO_4$ 0.6 g/l. Using this optimized fermentation method, the yield of laccase was increased 59.68 times to 67.258 U/ml compared with the laccase production with an unoptimized medium. This is the first report on the statistical optimization of laccase production by Trichoderma harzianum ZF-2.

• 한국생물공학회:학술대회논문집
• /
• 한국생물공학회 2005년도 생물공학의 동향(XVII)
• /
• pp.297-298
• /
• 2005

• Journal of Microbiology and Biotechnology
• /
• 제12권3호
• /
• pp.449-456
• /
• 2002

The objective of this study was to optimize medium composition of initial pH, tryptone, glucose, yeast extract, and mineral mixture for production of bacteriocin by Lactobacillus acidophilus ATCC 4356, using response surface methodology. A response surface approach including new statistical and plotting methods was employed for design and analysis of the experiment. An interiorly augmented central composite design was used as an experimental design. A normal-distribution log-link generalized linear model based on a subset fourth-order polynomial ($R^2$=0.94, Mean Error Deviance=0.0065) was used as an analysis model. This model was statistically superior to the full second-order polynomial-based generalized linear model ($R^2$=0.80, Mean Error Deviance=0.0140). Nonlinear programming determined the optimum composition of the medium as initial pH 6.35, typtone $1.21\%$, glucose $0.9\%$, yeast extract $0.65\%$, and mineral mixture $1.17\%$. A validation experiment confirmed that the optimized medium was comparable to the MRS medium in bacteriocin production, having the advantage of economy and practicality.

• Food Science and Biotechnology
• /
• 제18권6호
• /
• pp.1342-1350
• /
• 2009

This paper investigates the production and optimization of $\beta$-galactosidase enzyme using synthetic medium by Kluyveromyces lactis NRRL Y-8279 in stirred tank bioreactor. Response surface methodology was used to investigate the effects of fermentation parameters on $\beta$-galactosidase enzyme production. Maximum specific enzyme activity of 4,622.7 U/g was obtained at the optimum levels of process variables (aeration rate 2.21 vvm, agitation speed 173.4 rpm, initial sugar concentration 33.8 g/L, incubation time 24.0 hr). The optimum temperature and pH of the $\beta$-galactosidase enzyme produced under optimized conditions were $37^{\circ}C$ and pH 7.0, respectively. The enzyme was stable over a pH range of 6.0-7.5 and a temperature range of $25-37^{\circ}C$. The $K_m$ and $V_{max}$ values for O-nitrophenol-$\beta$-D-galactopyranoside (ONPG) were 1.20 mM and $1,000\;{\mu}mol/min{\cdot}mg$ protein, respectively. The response surface methodology was found to be useful in optimizing and determining the interactions among process variables in $\beta$-galactosidase enzyme production. Hence, this study fulfills the lack of using mathematical and statistical techniques in optimizing the $\beta$-galactosidase enzyme production in stirred tank bioreactor.

• Journal of Microbiology and Biotechnology
• /
• 제15권1호
• /
• pp.197-201
• /
• 2005

A novel chitinase-producing bacterial strain of Pantoea dispersa was isolated from the sea near Bhavnagar, India for efficient disposal of chitinous waste from the seafood processing industry. The medium components were optimized by using a cubic model in the central composite design for increasing chitinase production. The optimal concentrations for higher production of chitinase were (g l-1) chitin, 10.0; urea, 0.35; MgSO4 7H2O, 0.08, and CaCl2, 0.15. Here, peptone (0.05 g l-1) was used as a constant variant in all trials. Using a statistical optimization method, the chitinase production was found to increase from 108 to 486.4 units ml-1. Chitin was prepared from the crustacean waste, and Fourier Transform Infrared (FTIR) Spectroscopy was used to characterize the isolated chitin. Chitinous waste degradation was studied in terms of chitinase production.

• 한국미생물·생명공학회지
• /
• 제41권1호
• /
• pp.52-59
• /
• 2013

The nutritional requirements for the maximum production of lipopeptides by Bacillus subtilis N7 (B. subtilis N7) were investigated and optimized using response surface methodology (RSM) under shake flask fermentation. A one-factor-at-a-time experimental setup was used to screen carbon and nitrogen sources. A Plackett-Burman design (PBD) was employed to screen the most critical variables for lipopeptides production amongst ten nutritional elements. The central composite experimental design (CCD) was finally adopted to elucidate the composition of the fermentation medium. Statistical analyses (analysis of variance, ANOVA) of the results showed that KCl, $MnSO_4$ and $FeSO_4{\cdot}6H_2O$ were important components and that their interactions were strong. Lipopeptide production was predicted to reach 709.87 mg/L after a 60 h incubation using an optimum fermentation medium composed of glucose 7.5 g/L, peanut oil 1.25 g/L, $MgSO_4$ 0.37 g/L, $KH_2PO_4$ 0.75 g/L, monosodium glutamate 6.75 g/L, yeast extract and $NH_4Cl$ (5:3 w/w) 10 g/L, KCl 0.16 g/L, $FeSO_4{\cdot}6H_2O$ 0.24 mg/L, $MnSO_4$ 0.76 mg/L, and an initial pH of 7.0. Lipopeptide production ($706.57{\pm}3.70$ mg/L) in the optimized medium confirmed the validity of the predicted model.