scholarly journals Selection of Optimal Conditions for Cultivation of Bacillus megaterium UCM B-5710 – Producer of Keratinase

2021 ◽  
Vol 83 (6) ◽  
pp. 32-40
Author(s):  
K.V. Avdiyuk ◽  
◽  
A.O. Roy ◽  

Every year the volume of production of poultry products all over the world is growing steadily. This contributes to a constant increase in the amount of by-products of poultry processing in the form of down and feather waste, which are dangerous for the environment due to the hard-to-degrade keratin protein and a large number of microbial pathogens. Therefore, the use of environmentally friendly methods for the destruction of keratin substrates due to keratinases of microorganisms is an urgent area of research. The aim of this work was to select the optimal cultivation conditions for the Bacillus megaterium strain UCM B-5710 to increase the activity of the keratinase synthesized by it. Methods. The culture was grown at 28°C, 201 rpm for 7 days on a basic nutrient medium containing defatted chicken feathers as the only source of carbon and nitrogen. The selection of optimal cultivation conditions was carried out according to the following parameters: temperature (21°C, 28°C, 42°C), stirring speed (201 rpm, 212 rpm), amount of inoculum (5%, 10%, 15% , 20%, 25%), the initial pH value of the nutrient medium (4.0–11.0), concentration of keratin-containing substrate (0.1%, 0.2%, 0.5%, 1.0%, 1.5%, 2.0%), additional carbon source (glucose, galactose, lactose, maltose, sucrose, mannitol, potato and corn starch, soluble starch, soybean meal) and nitrogen (NH4Cl, NH4NO3, (NH4)2SO4, NaNO3, urea, peptone, tryptone, yeast extract and soybean meal) at a concentration of 1%. Keratinase activity was assessed by the UV absorption at 280 nm of the hydrolysis products of keratin-containing raw materials. Protein was determined by the Lowry method. Results. The dynamics of the enzyme synthesis showed that the culture of B. megaterium UCM B-5710 exhibited the highest keratinase activity on the 3rd day, and complete splitting of feathers was observed on the 4–5th days. The selection of the concentration of the keratin-containing substrate showed that 0.5% is the optimal concentration. The study of the influence of the initial pH value of the nutrient medium indicates that the culture grew well at pH 6.0–7.0 and pH 9.0–11.0, but at pH 8.0 its growth was very weak. The culture exhibited the maximum keratinase activity at pH 10.0. In addition, at this pH value, complete splitting of feathers was visually observed. The influence of such a key factor as temperature on the growth and synthesis of the enzyme by B. megaterium UCM B-5710 culture demonstrated complete splitting of feathers already on the 2nd day of cultivation at 42°C, at 21°C the culture split feathers very poorly. The introduction of the inoculum into the composition of the nutrient medium in an amount of 15% of the volume of the medium and the mixing intensity of 212 rpm turned out to be optimal. Besides, it was shown that the introduction of an additional source of carbon or nitrogen had an ambiguous effect on the level of keratinase activity of B. megaterium UCM B-5710. Complete inhibition of enzyme synthesis was observed when ammonium sulfate was added to the nutrient medium, and partial inhibition was observed in the case of glucose, lactose, and maltose. Potato, corn, and soluble starch stimulated keratinase synthesis. The majority of inorganic nitrogen sources (ammonium chloride and nitrate) did not affect the synthesis of B. megaterium UCM B-5710 keratinase, while organic sources (urea, peptone, tryptone, yeast extract) increased the level of keratinase activity by 20–50%. However, the most effective result was obtained using soybean meal, the addition of which to the nutrient medium increased the keratinase activity by 2.5 times. Conclusions. As a result of the studies, the optimal conditions for cultivation of the B. megaterium UCM B-5710 strain were selected: the optimum temperature for the growth and development of the culture is 42°C, the initial pH value is 10.0, the stirring speed is 212 rpm and the amount of inoculum introduced is 15%, an additional source of carbon and nitrogen in the form of soybean meal at a concentration of 0.5%. This made it possible to increase the activity of keratinase by 4 times.

Author(s):  
A. V. Malkova ◽  
I. Yu. Evdokimov ◽  
M. V. Shirmanov ◽  
A. N. Irkitova ◽  
D. E. Dudnik

Abstract: This article aims to develop a probiotic for animals and aquaculture based on the Bacillus toyonensis B-13249 and Bacillus pumilus B-13250 strains. The selection of a nutrient medium was conducted for cultivating the inoculum of these microorganisms. Several bacteria fermentations of the Bacillus genus were performed in biological reactors with a capacity of 15 and 250 l. A technology for obtaining a finished probiotic for animals and aquaculture was developed. The results indicate that L-broth is the most optimal nutrient medium for cultivating the studied strains. The cultivation of B. toyonensis B-13249 and B. pumilus B-13250 strains in fermenters revealed that sporulation begins after 4–8 hours of fermentation. In contrast to the vegetative medium, the fermentative medium helped the bacilli develop a higher optical density (the maximum value in the B. pumilus strain – 2.400±0.149), pH value (maximum value in the B. toyonensis strain – 8.483±0.609) and titer (at least 1010 CFU/g). After 20–24 hours of incubation, both strains of bacilli in the fermenter, almost completely pass into endospores, which serve as a signal for the start of biomass centrifugation. This was indicated by the following: from a 15 l fermenter – 83.3±6.1 g of concentrate, from a 250 l fermenter – 499.8±51.4 g. The number of bacilli in a concentrated state was at least 1·1011 CFU/g for both strains. Obtaining a finished preparation required mixing bacterial concentrates with maltodextrin to a titer of at least 1·1010 CFU/g. The number of bacteria in the preparation checked every month during the year, recorded no value less than 1·1010 CFU/g. Thus, L-broth is most favorable for growing the mother culture of the B. toyonensis B-13249 and B. pumilus B-13250 strains, and fermentative nutrient medium – for the cultivation in fermenters. The expiry date of the bacilli-based biological preparation is at least 12 months, during which the drug’s polycomponence, color and consistency are preserved, in addition to the bacteria titer (at least 1·1010 CFU/g) and their viability.


2021 ◽  
Vol 1 (2) ◽  
pp. 108-117
Author(s):  
Vanessa Jane Zainip ◽  
Liyana Amalina Adnan ◽  
Mohamed Soliman Elshikh

Industrial wastewater including dye waste disposal, has been released in a massive amount and is difficult to degrade, especially synthetic dyes. In this study, 10 different types of fungi were isolated from a decayed wood in UTM forest and were labelled as S1-S10. Two dyes were chosen for this study, which were Procion Red MX-5B (PRMX5B) and Remazol Brilliant Violet 5R (RBV5R). These fungi were screened for their ability to decolor both dyes and further tested for their ability to decolor the dyes in liquid medium under several parameters; carbon and nitrogen sources, initial pH value, temperature, and agitation. S1 decolorized PRMX5B efficiently with the addition of glucose (45%), ammonium nitrate (61%), pH 3 (69%), temperature 37°C (49%), and agitation 100 rpm (69%), whereas S2 decolorized efficiently with the addition of glucose (60%), ammonium nitrate (49%), pH 3 (70%), temperature 37°C (46%), and agitation 100 rpm (74%). S1 demonstrated efficient decolorization of RBV5R with the addition of glucose (80%), ammonium nitrate (62%), pH 3, temperature 37°C (75%), and agitation 100 rpm (90%), whereas S2 demonstrated efficient decolorization with the addition of glucose (52%), ammonium nitrate (67%), pH 3, temperature 37°C (75%), and agitation 100 rpm (71%).The percentage of decolorization of dyes was measured by using a UV-Vis spectrophotometer. These fungi were then identified using the 18sr RNA method. Based on macroscopic and microscopic characteristics and a polygenetic tree, fungi S1 belong to Trichoderma koningiopsis and fungi S2 belong to Trichoderma atroviride. 


2011 ◽  
Vol 343-344 ◽  
pp. 430-437 ◽  
Author(s):  
Ling Ling Zheng ◽  
Hai Jin Mou ◽  
Jing Li

This work presents laboratory studies on the degradation of lambda-cyhalothrin. At first, a rapid quantitative determination method of lambda-cyhalothrin in food was developed by high performance liquid chromatography. Lambda-cyhalothrin-degrading bacterium F37 was isolated from the sewage of a pesticide factory outlet and was identified as Citrobacter braakii. The effects of environmental factors including carbon and nitrogen sources, initial pH, medium volume, incubating temperature and substrate concentration on the degradation rate were investigated. The addition of sucrose and yeast extract at the concentrations of 4.0 and 3.0 g/L, respectively, was the best for the degradation of lambda-cyhalothrin. F37 showed higher degradation activity at the range of moderate pH value (pH 6.5-8.0). After 72-h stirring culture, the degradation rates of lambda-cyhalothrin reached 81.1% at pH 7.0. The degradation dynamics analysis showed that the degradation half-life times of lambda-cyhalothrin in the culture liquid of F37 were only 5.7, 1.9 and 4.9 days at pH 9.0, 7.0 and 5.0, respectively. In addition, cypermethrin and triazophos could also be degraded by F37, showing that F37 was a broad-spectrum pesticide- degrading bacterium. Application of F37 on eliminating pesticide in vegetable showed that 68% of lambda-cyhalothrin was removed after treatment for 48 h. The results indicated that Citrobacter braakii F37 is effective in the elimination of pesticide and may provide a potent application in detergent industry and environmental restoration.


Nanomaterials ◽  
2021 ◽  
Vol 11 (2) ◽  
pp. 529
Author(s):  
Hongjuan Zheng ◽  
Kongjun Zhu ◽  
Ayumu Onda ◽  
Kazumichi Yanagisawa

Eu(OH)3 with various shape-controlled morphologies and size, such as plate, rod, tube, prism and nanoparticles was successfully synthesized through simple hydrothermal reactions. The products were characterized by XRD (X-Ray Powder Diffraction), FE-SEM (Field Emission- Scanning Electron Microscopy) and TG (Thermogravimetry). The influence of the initial pH value of the starting solution and reaction temperature on the crystalline phase and morphology of the hydrothermal products was investigated. A possible formation process to control morphologies and size of europium products by changing the hydrothermal temperature and initial pH value of the starting solution was proposed.


2014 ◽  
Vol 79 (7) ◽  
pp. 815-828 ◽  
Author(s):  
Nikola Ilic ◽  
Slavica Lazarevic ◽  
Vladana Rajakovic-Ognjanovic ◽  
Ljubinka Rajakovic ◽  
Djordje Janackovic ◽  
...  

The sorption of inorganic arsenic species, As(III) and As(V), from water by sepiolite modified with hydrated iron(III) oxide was investigated at 25 ?C through batch studies. The influence of the initial pH value, the initial As concentrations, the contact time and types of water on the sorption capacity was investigated. Two types of water were used, deionized and groundwater. The maximal sorption capacity for As(III) from deionized water was observed at initial and final pH value 7.0, while the bonding of As(V) was observed to be almost pH independent for pH value in the range from 2.0 to 7.0, and the significant decrease in the sorption capacity was observed at pH values above 7.0. The sorption capacity at initial pH 7.0 was about 10 mg g?1 for As(III) and 4.2 mg g?1 for As(V) in deionized water. The capacity in groundwater was decreased by 40 % for As(III) and by 20 % for As(V). The Langmuir model and pseudo-second order kinetic model revealed good agreement with the experimental results. The results show that Fe(III)-modified sepiolite exhibits significant affinity for arsenic removal and it has a potential for the application in water purification processes.


1952 ◽  
Vol 30 (4) ◽  
pp. 360-370 ◽  
Author(s):  
Egon Stark ◽  
P. A. Tetrault

Thirty-five cultures of Bacillus stearothermophilus hydrolyzed five starches under various cultural conditions. Hydrolysis occurred regardless of the type, brand, or batch of starch; regardless of the initial pH or of the subsequent pH changes of the medium. Starch in broth was better attacked than in agar media. Some cultures hydrolyzed 0.5%, but not 1% starch; others hydrolyzed easily 10% soluble starch. Length of incubation was important. Certain cultures never formed acid or sugar from starch. Dextrinization was a more reliable indication of starch hydrolysis than was the formation of acid or sugar. Soluble starch gave more consistent results in repeated experiments than did nonsoluble starches. The type of protein medium determines strongly the formation of amylase. Trypticase was the best commercial medium, yeast extract came second. The other 10 media yielded fewer amylolytic cultures. Yeast extract added to media enhanced amylase formation, except with trypticase. Tryptose, proteose-peptone, and neopeptone inhibited the growth of most cultures.


2003 ◽  
Vol 21 (5) ◽  
pp. 451-462 ◽  
Author(s):  
Sameer Al-Asheh ◽  
Fawzi Banat ◽  
Leena Abu-Aitah

An improvement in the adsorption capacity of naturally available bentonite towards water pollutants such as Methylene Blue dye (MBD) is certainly needed. For this purpose, sodium bentonite was activated by two methods: (1) treatment with sodium dodecyl sulphate (SDS) as an ionic surfactant and (2) thermal treatment in an oven operated at 850°C. Batch adsorption tests were carried out on removing MBD from aqueous solution using the above-mentioned bentonites. It was found that the effectiveness of bentonites towards MBD removal was in the following order: thermal-bentonite > SDS-bentonite > natural bentonite. X-Ray diffraction analysis showed that an increase in the microscopic bentonite platelets on treatment with SDS was the reason behind the higher uptake of MBD. An increase in sorbent concentration or initial pH value of the solutions resulted in a greater removal of MBD from the solution. An increase in temperature led to an increase in MBD uptake by the bentonites studied in this work. The Freundlich isotherm model was employed and found to represent the experimental data well.


2013 ◽  
Vol 798-799 ◽  
pp. 1123-1127
Author(s):  
Hua Lei Zhou ◽  
Qiong Qiong Zhu ◽  
Dong Hua Huang

The activated carbon with high surface area was prepared by KOH activation from anthracite and used as adsorbent for removal of Cr (VI) from aqueous solution. The pore structure and surface properties were characterized by N2 adsorption at 77K, transmission electron microscope (TEM) and Fourier transform infrared spectroscopy ( FTIR). Effect of pH and isotherms at different temperature were investigated. Results show that the prepared carbon is a microporous-and mesoporous-adsorbent with developed pore structure and abundant surface oxygen-containing groups. PH value of the solution plays key function on the adsorption. The chemical adsorption dominates the adsorption process. The activated carbon exhibits much higher Cr adsorption capacity than the commercial activated carbon at initial pH of ~3. The equilibrium adsorption data are fitted by both Freundlich model and Langmuir model well.


2002 ◽  
Vol 20 (4) ◽  
pp. 393-416 ◽  
Author(s):  
Fawzi Banat ◽  
Sameer Al-Asheh ◽  
Dheaya‘ Al-Rousan

This study examined and compared the ability of chicken feathers, human hair and animal horns, as keratin-composed biosorbents, for the removal of Zn2+ and Cu2+ ions from single metal ion aqueous solutions under different operating conditions. The three biosorbents investigated in this study were all capable of adsorbing Zn2+ and Cu2+ ions from aqueous solutions. The biosorbent showing the highest uptake of Zn2+ and Cu2+ ions was animal horns. Chicken feathers showed a higher Cu2+ ion uptake and a lower Zn2+ ion compared to human hair. Increasing the initial concentration of Zn2+ or Cu2+ ions, or increasing the initial pH value, increased the metal ion uptake. Such uptake decreased when the temperature was raised from 25°C to 50°C for all adsorbent/metal ion combinations except for Zn2+ ion/human hair where the uptake increased with temperature. It was demonstrated that the addition of NaCl salt to the metal ion solution depressed the metal ion uptake. The Freundlich isotherm model was found to be applicable to the adsorption data for Cu2+ and Zn2+ ions.


Author(s):  
Zhao-Jun Wei ◽  
Le-Chun Zhou ◽  
Hua Chen ◽  
Gui-Hai Chen

Moranoline (1-Deoxynojirimycin, DNJ) is a piperidine alkaloid, and shows high inhibit activities to glucoamylase and ?-glucosidase. One DNJ high-yield strain of Streptomyces lawendulae was obtained after isolated form soil and mutated with the ultra violet (UV) and ethyl sulfate (DES), which named as TB-412, and can produce DNJ with 35.925 mg/L. Response surface methodology (RSM) was applied to optimize the parameters of DNJ yield from S. lawendulae TB-412. The effects of independent variables of fermentation, including time, temperature, initial pH and the soluble starch content were investigated. The statistical analysis showed that the fermentation time, pH and the soluble starch content, and the quadratics of time, temperature, pH and the soluble starch content, as well as the interactions between fermentation time and pH, and time and the soluble starch content, showed significant effects on DNJ yield. The optimal process parameters for DNJ production within the experimental range of the variables researched was at 11d, 27 °C, pH 7.5, and 8% soluble starch content. At this condition, the DNJ yield was predicted to be 42.875 mg/L.


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