[Optimization of culture condition of a moderately thermophilic acidophilic Ferroplasma thermophilum].
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Abstract:
The extreme microorganisms Ferroplasma spp., play an important role in bioleaching of sulphide ores at low pH value and temperatures around 50 degrees C. Without cell wall, Ferroplasma spp. is sensitive to pulp density, shearing force and heavy metal ions. Thus it is difficult to obtain their high cell density cultures, which limits the large-scale industrial application. In this paper, the optimum culture conditions of Ferroplasma thermophilum were studied by shaking culture. The results showed that the optimum culture conditions are as follows: 50 degrees C, initial pH 0.5, 50 mL working volume in 250 mL shaking-flask, inorganic nitrogen source (NH4)2SO4. The optimum combination of FeSO4.7H2O, yeast extract and peptone was determined by orthogonal experiments, including FeSO4.7H2O 40 g/L, yeast extract 0.3 g/L, peptone 0.2 g/L. Under the optimum culture conditions, the cell density was up to 6.3 x 10(7) cell/mL, and the oxidation of 40 g/L ferrous sulfate heptahydrate was finished in less than 72 hours. The results might provide information for scale-up of archaeon culture as well as its industrial application.Keywords:
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To improve the yield of the Pseudomonas strain N1 in the fermentation culture medium screened from the biofilm of BAF reactor, the optimal shake flask fermentation conditions for Pseudomonas N1 were determined by single factor test and orthogonal test. The composition of optimal fermentation culture medium was as follows: nitrate of potash of 4 g/L, sodium succinate of 26.1 g/L, KH 2 PO 4 of 4.0 g/L, MgSO 4 ·7H 2 O of 1 g/L, KCl of 1 g/L. The cell concen- tration could reach 9.08 g/L under the conditions. Fermentation was performed for 36h in a 250ml shake flask containing 70mL medium at 1% inoculums volume and initial pH value 7.0. The cell concentration could reach 12.86 g/L which was 2.5 fold higher than that under the original conditions. Simultaneous nitrification and denitrification could be achieved to alleviate the pollution of nitrogen in the wastewater by adding the microbial inoculums into the common aerobic nitrifica- tion sludge.
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Potassium nitrate
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We compared the effects of different carbon sources,nitrogen sources and inorganic salts on fermentation yield of oceanic red yeast(Rhodotorula sp.)RH1.By 4 factors and 3 levels orthogonal test,the optimum fermentation conditions for RH1 were determined: peptone 10 g·L-1,yeast extract 15 g·L-1,glucose 20 g·L-1,MgSO4 0.25 g·L-1,KH2PO4 0.25 g·L-1,NaCl 10 g·L-1,inoculation amount 10%,initial pH 5.0,temperature 28 ℃,80 mL medium in 500 mL triangle bottle,and fermentation time 48 h.Under above conditions,the cell biomass of RH1 can reach 10.46×108 cfu·mL-1,which increases by 23.9% as compared with the fermentation before optimization.An experiment of expanded culture is also conducted in a 25 L fermentation tank.The cell biomass of RH1 increased to 33.6×108 cfu·mL-1 in 28 h under the conditions of inoculation amount 8%,initial pH 5.0,revolution speed 350~400 r·min-1,air ventilation volume 9 L·min-1 and temperature 28 ℃.It is expected to further increase the biomass of strain RH1 by continuous filling of the materials.
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In this study the culture conditions and culture medium components of the Lactobacillus caseis-03 were optimized by means of single factor and orthogonal experiments.The results showed that Lactobacillus caseis-03 was anaerobic bacteria;the optimum culture conditions were as follows:initial pH 6.5,35 ℃,inoculation 1.0%,static cultured in improved MRS medium for 24 h;the optimum culture medium components were as follows:peptone 12 g,yeast extract 6 g,beef extract 6 g,glucose 18 g,sodium acetate 5 g,C6H17N3O7 2.15 g,Tween 80 1 mL,MgSO4·7H2O 0.58 g,MnSO4·4H2O 0.05 g,K2HPO4 2 g,H2O 1000 mL.D610 nm value could reach 1.832,viable bacteria counts could reach 3.22×1010 CFU/mL in the optimum culture medium and conditions.
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Optimum carbon source,nitrogen source and concentrations of inorganic salts were determined with single factor and orthogonal experiments.The optimum composition was as follows: saccharose 1.25%,soya peptone 1.25%,KH2PO3 2mmol/L,beef extract 0.3%,peptone 1%,NaCl 0.5%,Fe2(SO4)3 3mmol/L.The optimum incubation conditions were fermentation temperature 37℃,fermentation time 4d and inoculum 3% in 70ml medium.
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Liquid medium
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Isolation of thermophilic hydrogen bacteria was performed at 50°C using enrichment culture method. One of the four strains isolated, strain TH-1 grew most rapidly. Culture conditions of strain TH-1 were investigated. Optimum temperature and pH for growth proved to be 52°C and 7.0, respectively. There existed a positive correlation between the specific growth rate and the partial pressure of carbon dioxide in the gas phase. Ammonium and nitrate are the good nitrogen sources in that order. Effect of concentrations of nitrogen source, magnesium, ferrous and phosphate ions on the cell growth was also investigated. The maximum specific growth rate (μmax) of strain TH-1 was determined as 0.68 hr-1 by the cultivation at 52°C in a jar fermentor containing the optimal medium at pH 7.0.
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Objective:To get the highest sporulation rate and spore concentration, and offered the theoretical foundation for producing the Bacillus coagulans preparation on industry.Method: Influence of carbon sources,nitrogen sources,inorganic salts,Mn2+,initial pH,temperature,inoculum size and dissolved oxygen were investigated in shake flask or 15L auto fermentor.Result:The optimal medium was identified as follow: bran 20g/L,yeast extract 5g/L,soybean meal 10g/L, NaCl 5g/L,K2HPO4 3g/L,MnSO4·H2O 0.3g/L.The optimal culture conditions were as follows:initial pH 7.0,the initial spore concentration controlled at 106CFU/mL,40℃,180r/min,15mL medium in 250mL flask,dissolved oxygen level not less than 30% in the 15L auto fermentor,the spore concentration reached 5.8×109CFU/mL and sporulation rate reached 96.7% after cultivating for 20h.Conclusion: The results show that the optimal culture conditions can be applied in practical production.
Bacillus coagulans
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A kind of strain with high flocculent activity named as TJ-30 was separated and screened from the activated sludge in Tangjiatuo sewage treatment plant of Chongqing.Orthogonal experiment and even experiment were used to obtain the culture conditions and the compositions of the culture medium for this strain,and the results showed that the optimum carbon source for this strain was soluble starch,its nitrogen source was yeast extract,its culture pH was acidic,and its aeration was cotton plug.Its optimum culture medium composition was:soluble starch 25.252 2g,yeast extract 0.1 g,K2HPO4 0.778 2 g,KH2PO4 6.846 8 g,urea 0.008 9 g,(NH4)2SO4 0.739 7 g,NaCl 0.1g,H2O 1 000 mL.Flocculation rate of this strain changeed with the change of the culture days and reached stable period after 40 hours.The flocculation activity of this strain on kaolin suspension was over 80% after optimization of the culture conditions.
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Yeast extract
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Optimization on the Screening and Culture Condition of Biosurfactant-producing Bacteria from Compost
[Objective] The study aimed to screen out piosurfactant-producing bacteria from compost and optimize its culture condition.[Method] The strains which could produce biosurfactant were screened from the agriculture aerobic compost samples with methods of enrichment culture and strain purification ect.and the cultural condition of strain was optimized through orthogonal test.[Result] A strain BS-2 that could produce biosurfactant was screened out from the agriculture aerobic compost,which could decreased the surface tension of broth below 40 mN/m and keep the same level under the temperature of 20-100 ℃ and pH of 6.0-9.5, showing its good surface tension and adaptability in composting environment.The optimized fermentation conditions were as follows: soluble starch 25.0 g/L,NH4NO3 8.0 g/L,KH2PO4 2.0 g/L,K2HPO4 2.5 g/L,KCl 1.1 g/L,NaCl 1.1 g/L,MgSO4 0.15 g/L,FeSO4·7H2O 5.0×10-5 g/L,EDTA 1.0 g/L,yeast paste 0.2 g/L,initial pH of 7.0,temperature of 30 ℃,rotation speed of 150 r/min,culture time of 3 d.Under this condition,the lowest surface tension of the broth was measured as 29.3 mN/m.[Conclusion] The strain BS-2 was preliminarily identified as Bacillus subtilis.
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Enrichment culture
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According to flask experiments,the culture conditions of D-lactic acid production by Serratia marcescens are determined as follows:culture temperature of 34 ℃,pH value of 6.5,the inoculum amount of 5%,loading liquid of 150 mL/500 mL,two-stage(pre-aerobic culture to accumulated the thallus,later anaerobic fermentation) culture method.Effects of medium components on cell growth and lactic acid yield are studied.Glucose is selected as carbon source,yeast is selected as nitrogen source,and meanwhile appropriate metal ions are added into medium.The optimal medium components(g·L-1)are as follows:glucose 15,yeast 15,KH2PO4 1.5,K2HPO4·3H2O 2.0,MgSO4·7H2O 1.0,FeSO4 ·7H2O 0.03,MnSO4·H2O 0.005.Under the optimal culture conditions and medium components by shake flask fermentation,D-lactic acid yield increases from(13.5±1.29) g·L-1 to(29.0±1.42) g·L-1 with optical purity over 97%.
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The phenol-degrading strain named JF-6,which is screened from activation sludge,can grow in inorganic salt medium with 1800 mg·L-1 phenol.The culture conditions are investigated by single factor experiment.The results show that the degradation rate of phenol reaches 100% at phenol concentration of 1000 mg·L-1.The optimum culture conditions are obtained as follows: temperature 20~40℃,pH value 4~9,NaCl concentration≤4.0%,DO(broth content) 40~100 mL of 250 mL shake flask,with peptone as additional nitrogen source.
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Degradation
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