Optimization Of Fermentation Medium Research Articles

Decoupling production from growth by magnesium sulfate limitation boosts de novo limonene production

The microbial production of isoprenoids has recently developed into a prime example for successful bottom-up synthetic biology or top-down systems biology strategies. Respective fermentation processes typically rely on growing recombinant microorganisms. However, the fermentative production of isoprenoids has to compete with cellular maintenance and growth for carbon and energy. Non-growing but metabolically active E. coli cells were evaluated in this study as alternative biocatalyst configurations to reduce energy and carbon loss towards biomass formation. The use of non-growing cells in an optimized fermentation medium resulted in more than fivefold increased specific limonene yields on cell dry weight and glucose, as compared to the traditional growing-cell-approach. Initially, the stability of the resting-cell activity was limited. This instability was overcome via the optimization of the minimal fermentation medium enabling high and stable limonene production rates for up to 8 h and a high specific yield of ≥50 mg limonene per gram cell dry weight. Omitting MgSO4 from the fermentation medium was very promising to prohibit growth and allow high productivities. Applying a MgSO4 -limitation also improved limonene formation by growing cells during non-exponential growth involving a reduced biomass yield on glucose and a fourfold increase in specific limonene yields on biomass as compared to non-limited cultures. The control of microbial growth via the medium composition was identified as a key but yet underrated strategy for efficient isoprenoid production. Biotechnol. Bioeng. 2016;113: 1305-1314. © 2015 Wiley Periodicals, Inc.

Effect of Solid State Fermentation Medium Optimization on Pleurotus ostreatus Laccase Production.

The objective of this work was to increase laccase production by Pleurotus ostreatus PLAB through culture medium optimization using solid state culture conditions. Increased laccase activity was obtained through design of experiments (DOE) using the Taguchi orthogonal array (OA). Seven factors, viz. lignocellulose, glucose, yeast extract, peptone, KH(2)PO(4), MgSO(4) 7H(2)O and MnSO(4) H(2)O at three levels and pH at two levels. OA layout of L18 (2(1) x 3(7)) was selected for the proposed experimental design using Minitab 17 software. Data analysis showed that lignocellulose (20 %) and glucose (10 g L1) had positive effect, whereas KH(2)PO(4), MgSO(4)∙7H(2)O and MnSO(4)∙H(2)O did not have significant effect on laccase production. Taguchi OA analysis showed that pH 6, lignocellulose 20 %, glucose 10 g L(-1), yeast extract 6 g L(-1), peptone 15 g L(-1), KH(2)PO(4) 3 g L1, MgSO(4)∙7H(2)O 0.5 g L(-1) and MnSO(4)∙H(2)O 0.1 g L-1 were the optimal conditions to maximize laccase production. The model predicted a 30.37 U g(-1) dry wt., which agreed with the experimentally obtained laccase activity 29.15 U g(-1) dry wt. at optimal conditions.

Improved submerged Aspergillus ficuum phytase production in bench-top bioreactors by optimization of fermentation medium

Phytase is an important feed and food additive, which is used in diets to increase the absorption of divalent ions, amino acids, and proteins in the bodies and to decrease the excessive phosphorus release in the manure to prevent negative effects on the environment. To date, phytase has been mostly produced in solid state fermentations with insignificant production volumes. Thus, there is a need to produce phytase in submerged fermentations, which can be scaled-up for commercial productions. Additionally, optimization of fermentation medium has not been studied well in the literature. Therefore, this study has been undertaken to improve Aspergillus ficuum phytase production in submerged fermentations by optimizing important nutrients in the fermentation medium (glucose, Na-phytate, and CaSO4) using Box-Behnken design of Response Surface Methodology. Also, effects of pH and temperature on phytase activity were studied. Optimum glucose, Na-phytate, and CaSO4 concentrations were determined as 126, 14, and 1.1 g l–1, respectively. Additionally, pH 5.5 and 55 ºC were determined as optimum for the produced A. ficuum phytase activity. Under these conditions, phytase activity was increased to 3.45 U ml–1, which is about 50% higher than the previous results. Furthermore, the lowest activity loss was observed under 4 ºC storage conditions during 1 week of storage.

OPTIMASI MEDIA FERMENTASI Aspergillus oryzae, PENGHASIL ANTIJAMUR PATOGEN BUAH KAKAO Phytophthora palmivora Fermentation Medium Optimization of Aspergillus oryzae, Antifungals Producer for Cacao Pathogen Phytophthora palmivora

Indonesia was known as a producer and exporter of cacao, but nowadays Indonesia had an issue in cacao production caused by microorganisms (Phytophtora palmivora) that capable of reducing the amount of cacao production each year. This study follows up a previous study that had found compounds that are actively against at Phytophtora palmivora from Aspergillus oryzae metabolites. In this study the compound will be produced in greater numbers to optimize fermentation medium. The first stage was conducted to determine the profile of the active compound and to analyze the best carbon, nitrogen source for Aspergillus oryzae fermentation medium. The second stage was designed by using Response Surface Method (RSM) with Central Composite Design (CCD) to optimize two variables in the fermentation medium, glucose concentration (the best carbon source) and the concentration of peptone (best nitrogen source). The expected response was the most active compounds percentage of the area. Data analysis was performed by Design Expert DX 7.1.5 program. The first stage of this study presumed that Tenuazonic acid compounds with a molecular weight of 198.1126 gram.mol was an antifungal compound produced by Aspergillus oryzae fungus to prevent Phythopthora palmivora fungus pathogens cacao pods. In addition the results are that glucose was the best carbon source (mean % relative area of antifungal compounds: 85.39%) and that peptone was selected as the best source of nitrogen (mean % relative area of antifungal compounds: 91.07%). The second stage showed the optimum conditions of fermentation medium antifungal Aspergillus oryzae was the composition of glucose 35.25 g/L and peptone 16.7 g/L. The combination of both compositions would yield a mean response of laboratory data % relative area of antifungal compounds 91.2217% and the estimated value (prediction) by Design Expert models 92.2936%. Difference in the estimated values (the response of the model) to the value of the results of response in laboratory experiments at 1.1614%, then it could be concluded that the model used was appropriate and able to explain the applied experimentaldata.-1Kata kunci: Cacao, Aspergillus oryzae, Phytophtora palmivora, active compound, optimization of fermentation medium, Response Surface Method (RSM)

Response Surface Methodology as a StatisticalTool for Fermentation Media Optimization inLipase Production by Palm Oil Mill Effluent(POME) Isolate Emericella Nidulans NFCCI 3643

Among enzymes, lipases have been attracting biotechnologists owing to their numerous applications in various industries like – food, pharmaceutical, detergent, dairy, cosmetic, textiles etc. In the present study, optimization of lipase production by Emericella nidulans NFCCI 3643 was carried out using statistical experimental designs. Initially, a Plackett-Burmen design (PBD) was employed to find out the significant process variables. The results indicated that olive oil, ammonium sulphate, incubation period, temperature and pH had significant influences on lipase production. The optimal conditions were further obtained by subsequently using Box-Behnken design under response surface methodology (RSM). The interactive effects of 5 variables on lipase production were studied in 46 experimental runs and the optimum concentrations of these parameters were found to be olive oil, 18.98 mL/L; ammonium sulphate, 14.38 g/L; incubation time, 3.66 days; temperature, 25.71 °C and pH, 5.17. Under these optimal conditions the lipase activity was found to be 409.723 U/ml, which is a 2.76-fold increase in comparison with that obtained by optimization using traditional OFAT (one factor at a time) method.

Optimization of Fermentation Medium for Glycyrrhizin Biotransformation to Monoglucuronyl-glycyrrhetinic Acid by Plackett-Burman and Box-Behnken Design

Plackett-Burman (PB) and Box-Behnken (BB) experimental designs were used to optimize fermentation variables for the biotransformation of glycyrrhizin (GL) to monoglucuronyl-glycyrrhetinic acid (MGGA). The PB design was first used to screen the important factors among the medium variables. The steepest ascent method was used to approach the optimum range for each of these factors. The BB design was finally used to analyze the response surfaces of the screened factors for further optimization. The optimized conditions for this system were 0.7 g/L MgSO4·7H2O, 1.19 g/L GL, and cultivation for six days. The biotransformation of GL to MGGA could reach up to 35.72%, which is a good result for this kind of transformation.

Fermentation optimization for the production of bioactive polysaccharides from Cordyceps sinensis fungus UM01

The optimal fermentation conditions and medium for the production of bioactive polysaccharides from the mycelium of Cordyceps sinensis fungus UM01 were investigated by using orthogonal design and high performance size exclusion chromatography coupled with multi-angel laser light scattering and refractive index detector (HPSEC-MALLS-RID). Results showed that the optimal temperature, initial pH, rotation speed, medium capacity (ratio of medium volume to the volume of flask bottle) and inoculums volume for the mycelium growth were 15°C, pH 6.0, 150rpm, 2/5 (v/v), and 3% (v/v), respectively. Furthermore, bioactive polysaccharides from the mycelium of C. sinensis fungus UM01 were determined as polysaccharide fractions with the molecular weight above 10kDa. The optimal fermentation medium was determined as a composition of glucose 30.0g/L, sucrose 30.0g/L, KH2PO4 1.0g/L, CaCl2 0.5g/L, yeast extract 3.0g/L, and MgCl2 0.1g/L according to the maximum amount of the bioactive polysaccharides (486.16±19.60mg/L) measured by HPSEC-MALLS/RID. Results are helpful to establish an efficient and controllable fermentation process for the industrial production of bioactive polysaccharides from C. sinensis UM01, and beneficial to develop a unique health and functional product in future.

A thermo‐alkaline lipase from a new thermophile Geobacillus thermodenitrificans AV‐5 with potential application in biodiesel production

AbstractBACKGROUNDA thermophilic lipase‐producing Geobacillus thermodenitrificans strain AV‐5 was isolated from the Mushroom Spring of Yellowstone National Park in WY, USA and studied as a source of lipase for transesterification of vegetable oils to biodiesel.RESULTSA maximum activity of 330 U mL−1 was produced on 2% (v/v) waste cooking oil at 50 °C, pH 8, aeration rate of 1 vvm and agitation speed of 400 rpm. However, the higher lipase productivity (14.04 U mL−1 h−1) was found at a volumetric oxygen transfer coefficient (kLa) value of 18.48 h−1. The partially purified lipase had a molecular weight, temperature and pH optimum of 50 kDa, 65 °C and pH 9, respectively, and was thermo‐alkali stable: at 70 °C, it retained 81% activity and 45% stability; at pH 10 it lost only 15% and 2.6% of its maximum activity and stability, respectively. Enzyme kinetic studies with p‐nitrophenyl laurate as substrate revealed high substrate specificity (km of 0.440 mmol L−1) and kinetic activity (vmax of 556 nmol mL min−1) of lipase.CONCLUSIONSThe kLa was found to be highly dependent on aeration and agitation rates. Following optimization of fermentation medium and parameters, a 7.5‐fold increase in lipase production by G. thermodenitrificans was attained. The lipase activity and substrate specificity (as km) are among the highest reported in the literature for bacterial lipases. It was demonstrated that the enzyme can produce biodiesel from waste cooking oil with a conversion yields of 76%. © 2015 Society of Chemical Industry

Enhanced production of dimethyl phthalate-degrading strain Bacillus sp. QD14 by optimizing fermentation medium

article i nfo Background: Integrated statistical experimental designs were applied to optimize the medium constituents for the production of a dimethyl phthalate (DMP)-degrading strain Bacillus sp. QD14 in shake-flask cultures. A Plackett-Burman design (PBD) was applied to screen for significant factors, followed by the Steepest Ascent Method (SAM) to find the nearest region of maximum response. A Box-Behnken design (BBD) of the Response Surface Methodology (RSM) was conducted to optimize the final levels of the medium components. Results: After the regression equation and response surface contour plots were analyzed, the concentrations of glucose, corn meal and NaCl were found to significantly influence the biomass of DMP-degrading bacteria. A combination of 22.88 g/L of glucose, 11.74 g/L of corn meal, and 10.34 g/L of NaCl was optimum for maximum biomass production of Bacillus sp. QD14. A 57.11% enhancement of the biomass production was gained after optimization in shake-flask cultivation. The biomass production of Bacillus sp. QD14 reached 9.13 ± 0.29 × 10 8 CFU/mL, which was an excellent match for the predicted value, and the mean value of the match degree was as high as 99.30%. Conclusion: In this work, the key factors affected by the fermentation of DMP-degrading strain Bacillus sp. QD14 were optimized by PBD, SAM and BBD (RSM); the yield was increased by 57,11% in the conditions in our study. We propose that the conditions optimized in the study can be applied to the fermentation for commercialization production.

Statistical experimental design optimization of rhamsan gum production by Sphingomonas sp. CGMCC 6833.

Rhamsan gum is a type of water-soluble exopolysaccharide produced by species of Sphingomonas bacteria. The optimal fermentation medium for rhamsan gum production by Sphingomonas sp. CGMCC 6833 was explored definition. Single-factor experiments indicate that glucose, soybean meal, K(2)HPO(4) and MnSO(4) compose the optimal medium along with and initial pH 7.5. To discover ideal cultural conditions for rhamsan gum production in a shake flask culture, response surface methodology was employed, from which the following optimal ratio was derived: 5.38 g/L soybean meal, 5.71 g/L K(2)HPO(4) and 0.32 g/L MnSO(4). Under ideal fermentation rhamsan gum yield reached 19.58 g/L ± 1.23 g/L, 42.09% higher than that of the initial medium (13.78 g/L ± 1.38 g/L). Optimizing the fermentation medium results in enhanced rhamsan gum production.

Industrial vitamin B12 production by Pseudomonas denitrificans using maltose syrup and corn steep liquor as the cost-effective fermentation substrates.

The aerobic Pseudomonas denitrificans is widely used for industrial and commercial vitamin B12 fermentation, due to its higher productivity compared to the anaerobic vitamin B12-producing microorganisms. This paper aimed to develop a cost-effective fermentation medium for industrial vitamin B12 production by P. denitrificans in 120,000-l fermenter. It was found that maltose syrup (a low-cost syrup from corn starch by means of enzymatic or acid hydrolysis) and corn steep liquor (CSL, a by-product of starch industry) were greatly applicable to vitamin B12 production by P. denitrificans. Under the optimal fermentation medium performed by response surface methodology, 198.27 ± 4.60 mg/l of vitamin B12 yield was obtained in 120,000-l fermenter, which was close to the fermentation with the refined sucrose (198.80 mg/l) and was obviously higher than that obtained under beet molasses utilization (181.75 mg/l). Therefore, maltose syrups and CSL were the efficient and economical substrates for industrial vitamin B12 fermentation by P. denitrificans.

Screening of Actinomycetes for Enzyme and Antimicrobial Activities from the Soil Sediments of Northern Tamil Nadu, South India

The present study involves the screening of actinomycetes for enzymes and antimicrobial activities from the soil sediments of Northern Tamil Nadu, South India. Actinomycetes from the soil samples were isolated by pour plate technique on Actinomycetes Isolation Agar. Morphological characteristics of the isolates were studied based on microscopic appearance. Actinomycetes isolates was screened for the production of different extracellular enzymes like amylase, protease and lipase. In the screening for enzyme activity, the actinomycetes strains showed amylase activity, protease activity and lipase activity. Preliminary screening for antimicrobial activity of actinomycetes isolates were performed by cross streak plate method. Active form of 82 isolated strains showed activity against one or more bacteria and exhibited potential antifungal activity. The active strain was subjected to cultural, molecular identification and optimization of fermentation medium, solvent extraction and antimicrobial activity for crude extracts. Based on the primary antimicrobial proficiency, Loyola AR1 strain was selected for further studies. Ethyl acetate extract showed good antimicrobial activity when compared to butanol extract. The partial 16s rRNA sequence analysis of Loyola AR1 isolate identified to be Streptomyces sp. and the sequence was submitted to the Gene Bank (HM163569).

Physiological studies on a locally isolated rhamnolipid producer, Pseudomonas strain

Aim Bacterial strains were isolated from different soil samples and evaluated for their ability to produce biosurfactants. Preliminary identification was made of the selected strain Kh4, as well as of the type of produced biosurfactant. Finally, the most optimum conditions for improving growth and the percentage of reduction in the culture surface tension (ST) were investigated. Materials and methods The genus of the selected strain was preliminarily identified on the basis of some biological tests and characterized pigmentation on a specific medium. Rhamnolipid production was confirmed using both Siegmund Wagner and Dubois methods as well as thin-layer chromatography. Quantitative measurements of the ST of cell-free supernatants were determined using the Du Nouy ring method in order to estimate the amount of biosurfactant produced. Cultivation conditions were studied on the basis of both flask and 7 l stirred tank bioreactor levels. Results A total of 66 bacterial strains were isolated from Egyptian soil and assessed for biosurfactant productivity. The selected isolate, preliminarily identified as a Pseudomonas spp., has the ability to produce rhamnolipid and hence to decrease the ST of the fermentation medium from 60 to about 35 mN/m. The optimum fermentation medium, used for the growth and maximum decrease in ST, was determined. Some physiological parameters such as the initial pH, incubation temperature, and time were also studied. Furthermore, the effect of pH control during the fermentation production of rhamnolipid in a 7 l stirred bioreactor was also studied. Accordingly, a maximum decrease in the ST of the cell-free culture medium of 32 mN/m was reached, confirming the production of a satisfactory level of rhamnolipid biosurfactant. Conclusion These results qualified the locally isolated Pseudomonas spp. Kh4 for use in the bioremediation of hydrocarbon-contaminated sites as well as in petroleum industries.

Optimization of Fermentation Medium for Extracellular Lipase Production from Aspergillus niger Using Response Surface Methodology.

Lipase produced by Aspergillus niger is widely used in various industries. In this study, extracellular lipase production from an industrial producing strain of A. niger was improved by medium optimization. The secondary carbon source, nitrogen source, and lipid were found to be the three most influential factors for lipase production by single-factor experiments. According to the statistical approach, the optimum values of three most influential parameters were determined: 10.5 g/L corn starch, 35.4 g/L soybean meal, and 10.9 g/L soybean oil. Using this optimum medium, the best lipase activity was obtained at 2,171 U/mL, which was 16.4% higher than using the initial medium. All these results confirmed the validity of the model. Furthermore, results of the Box-Behnken Design and quadratic models analysis indicated that the carbon to nitrogen (C/N) ratio significantly influenced the enzyme production, which also suggested that more attention should be paid to the C/N ratio for the optimization of enzyme production.

Optimization of Fermentation Medium for the Production of Recombinant Human Antithrombin III from Saccharomyces cerevisiae through Statistical Experimental Designs

Statistical Optimization of Olanzapine Ternary Solid Dispersions with Pvp K 30 and Peg 20,000 by Response Surface MethodologyAnil Katharia, Ratendra Kumar, Rajiv Sharma, Yogendra Singh, Uday Veer Singh Teotia

Optimization of Streptomyces sp.A11 Medium Cultivation on Cyclo(Tyrosyl-Prolyl) Production Using the Response Surface Methodology

The Response Surface Methodology is a three factorial design which provides the relationship between one or more measured dependent response and a number of input (independent) factors. Response surface methodology was used for optimization of fermentation medium for antibacterial agent cyclo(tyr-pro) production by Streptomyces sp. A11. The optimal response region of the significant factor was predicted by using a second order polynomial model fitted to the results obtained by applying the Central Composite Design (CCD) statistical design. Determination of the best production medium of cyclo(tyr-pro) showed among 6 carbon sources used, dextrin produced the highest cyclo(tyr-pro). Among the five nitrogen sources used, peptone produced the highest cyclo(tyr-pro), and the mineral salts group I produced the highest cyclo(tyr-pro) compared with others. The results of optimization showed that three variables (dextrin as a carbon source, peptone as nitrogen source, and mixture of mineral salts) used showed significantly affected to the activity of cyclo(tyr-pro). The mathematical model used, obtained by the composition of the three variables most optimum were concentration of dextrin was 32.55 g.L-1 , peptone concentration was 11.22 g L-1 , and 8.65 mL of mineral salts. By using the composition of mathematic model, cyclo(tyr-pro) was obtained 51.54 mg L-1 (model expectations) and 50.04 mg L-1 from experiments. The differences of experiment with expectation response value was 2.91%. Using the optimized parameters, the studies demonstrated an increase in the cyclo(tyr-pro) was 2.5 fold from 20 mg L-1 to 50 mg L-1

Transgalactosylating β-galactosidase from probiotic Lactobacillus plantarum MCC2156: production and permeabilization for use as whole cell biocatalyst.

Key nutritional factors were optimized for the maximum production of transgalactosylating β-galactosidase from Lactobacillus plantarum MCC2156. Galactose, yeast extract, sodium acetate and manganese sulphate were the most important nutrients affecting β-galactosidase production. Maximum β-galactosidase production (3015 miller units) was obtained by culturing L. plantarum in the optimized fermentation medium containing (w/v) galactose (4%), yeast extract (2%), sodium acetate (3%) and manganese sulphate (0.075%) with an optimum medium pH of 7.0, after 14h of incubation at 35°C. Further, permeabilization of L. plantarum cells using various chemical/ solvents for maximum β-galactosidase activity was performed for use as whole cell biocatalyst. Mixture of ethanol: n-butanol was found to effectively permeabilize the cells with maximum β-galactosidase activity under the following optimum conditions; 1: 1 mixture of ethanol (10%, v/v): n-butanol (30%, v/v) with a contact time of 10min at 28 ± 2°C.

Optimization of Fermentation Conditions for Cellulase Production by HDZK-BYTF620 (<i>Aureobasidium pullulans</i>) which is from the Gut of Termites

The components of a fermentation medium were optimized to produce cellulose from HDZK-BYTF620 (Aureobasidium pullulans) which is from the gut of termites and can produce cellulose under liquid state fermentation conditions by response surface methodology. By Plackett-Burman design screened out three major factors of strains HDZK-BYTF620 which affected production of the activity cellulose, immediately, CaCl2, MgSO4and CMC-Na. The fermentation conditions were optimized with Box-Behnken experiment and the largest regional of the activity of cellulase was approach with the steepest ascent experiment. The optimum composition of medium addition of CaCl2, MgSO4and CMC-Na was 0.77 g/L, 0.41 g/L and 21.88 g/L, respectively. Under the optimum fermentation medium conditions, the activity of cellulase was (598.00±2.56)×10-3IU/mL, increased 5.0 % to that of using initial medium.

Enhanced Production of Vitamin K2 from Bacillus subtilis (natto)by Mutation and Optimization of the Fermentation Medium

The aim of this study was to enhance the production of vitamin K2 by using N-methyl-N-nitro-N-nitroso-guanidine (NTG) and low energy ion beam implantation and optimizing the fermentation medium. Mutation resulted in 1.66-fold higher production of vitamin K2 than that of the parentl strain. The production by the mutant BN-P15-11-1was increased 55% and reached 3.593±0.107 mg/L by using the Plackett-Burman and Box-Behnken designs to optimize the fermentation medium. The optimal fermentation culture medium was composed of (g/L) glycerol 69.6, sucrose 34.5, K2HPO4 4.0, peptone 20, yeast extract 25 and fermented at 37 °C and 150 rpm for 72 h. The results showed that the NTG and low energy ion beam implantation mutations and optimizing fermentation medium were effective methods to enhance vitamin K2 production.

Enhanced Production of Vitamin K2 from Bacillus subtilis (natto) by Mutation and Optimization of the Fermentation Medium

The aim of this study was to enhance the production of vitamin K2 by using N-methyl-N-nitro-N-nitroso-guanidine (NTG) and low energy ion beam implantation and optimizing the fermentation medium. Mutation resulted in 1.66-fold higher production of vitamin K2 than that of the parentl strain. The production by the mutant BN-P15-11-1was increased 55% and reached 3.593±0.107 mg/L by using the Plackett-Burman and Box-Behnken designs to optimize the fermentation medium. The optimal fermentation culture medium was composed of (g/L) glycerol 69.6, sucrose 34.5, K2HPO4 4.0, peptone 20, yeast extract 25 and fermented at 37 °C and 150 rpm for 72 h. The results showed that the NTG and low energy ion beam implantation mutations and optimizing fermentation medium were effective methods to enhance vitamin K2production.