Isolation and Optimization of Culture Conditions of a Bioflocculant-Producing Fungi from Kombucha Tea SCOBY

Biolocculants are gaining attention in research due to their environmental friendliness and innocuousness to human in comparison to the conventional flocculants. The present study aimed to investigate the ability of fungi from Kombucha tea SCOBY to produce effective bioflocculant in bulk. A 16S rRNA gene sequence analysis was utilized to identify the isolate. The medium composition (carbon and nitrogen sources) and culture conditions (inoculum size, temperature, shaking speed, pH, and time) were optimized using one-factor-at-a-time method. The functional groups, morphology, and crystallinity of the bioflocculant were evaluated using Fourier transform infrared (FT-IR), scan electron microscope (SEM) and X-ray diffractometry (XRD). The fungus was found to be Pichia kudriavzevii MH545928.1. It produced a bioflocculant with flocculating activity of 99.1% under optimum conditions; 1% (v/v) inoculum size, glucose and peptone as nutrient sources, 35 °C, pH 7 and the shaking speed of 140 rpm for 60 h. A cumulus-like structure was revealed by SEM; FT-IR displayed the presence of hydroxyl, carboxyl, amine, and thiocynates. The XRD analysis demonstrated the bioflocculant to have big particles with diffraction peaks at 10° and 40° indicating its crystallinity. Based on the obtained results, P. kudriavzevii MH545928.1 has potential industrial applicability as a bioflocculant producer.

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Isolation, Identification and Characterization of Bioflocculant-Producing Bacteria from Activated Sludge of Vulindlela Wastewater Treatment Plant

Applied Microbiology ◽

10.3390/applmicrobiol1030038 ◽

2021 ◽

Vol 1 (3) ◽

pp. 586-606

Author(s):

Nkanyiso Celukuthula Nkosi ◽

Albertus K. Basson ◽

Zuzingcebo G. Ntombela ◽

Tsolanku S. Maliehe ◽

Rajasekhar V. S. R. Pullabhotla

Keyword(s):

Activated Sludge ◽

Treatment Plant ◽

16S Rrna Gene Sequencing ◽

Industrial Applications ◽

Culture Conditions ◽

Inoculum Size ◽

Medium Composition ◽

Rrna Gene ◽

Sequencing Analysis ◽

Optimal Medium

The low microbial flocculant yields and efficiencies limit their industrial applications. There is a need to identify bacteria with high bioflocculant production. The aim of this study was to isolate and identify a bioflocculant-producing bacterium from activated sludge wastewater and characterise its bioflocculant activity. The identification of the isolated bacterium was performed by 16S rRNA gene sequencing analysis. The optimal medium composition (carbon and nitrogen sources, cations and inoculum size) and culture conditions (temperature, pH, shaking speed and time) were evaluated by the one-factor-at-a-time method. The morphology, functional groups, crystallinity and pyrolysis profile of the bioflocculant were analysed using scanning electron microscope (SEM), Fourier transform infrared (FTIR) and thermogravimetric (TGA) analysis. The bacterium was identified as Proteus mirabilis AB 932526.1. Its optimal medium and culture conditions were: sucrose (20 g/L), yeast extract (1.2 g/L), MnCl2 (1 g/L), pH 6, 30 °C, inoculation volume (3%), shaking speed (120 rpm) for 72 h of cultivation. SEM micrograph revealed the bioflocculant to be amorphous. FTIR analysis indicated the presence of hydroxyl, carboxyl and amino groups. The bioflocculant was completely pyrolyzed at temperatures above 800 °C. The bacterium has potential to produce bioflocculant of industrial importance.

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Growth morphology of Streptomyces akiyoshiensis in submerged culture: influence of pH, inoculum, and nutrients

Canadian Journal of Microbiology ◽

10.1139/m92-016 ◽

1992 ◽

Vol 38 (2) ◽

pp. 98-103 ◽

Cited By ~ 20

Author(s):

M. A. Glazebrook ◽

L. C. Vining ◽

R. L. White

Keyword(s):

Nitrogen Sources ◽

Growth Conditions ◽

Culture Conditions ◽

Inoculum Size ◽

Clear Correlation ◽

Growth Morphology ◽

Physiological Control ◽

Submerged Cultures ◽

Pellet Formation ◽

Culture Influence

Most media in which the growth of shaken submerged cultures of Streptomyces akiyoshiensis was examined did not support the formation of well-dispersed mycelial suspensions. Investigation of the culture conditions promoting dispersed growth showed the pH of the culture medium to be of critical importance; an initial value of 5.5 minimized aggregation of the mycelium while supporting adequate biomass production. In cultures started at this pH, spore inocula gave better mycelial dispersal than did vegetative inocula; with spore inocula, growth morphology was also less affected by inoculum size. The composition of the nutrient solution influenced the extent of mycelial dispersal; slow growth was often associated with clumping but no clear correlation was observed between pellet formation and the ability of carbon or nitrogen sources to support rapid growth. Increasing the phosphate concentration from 0.5 to 15 mM caused a modest decrease in mycelial aggregation. Conditions promoting a well-dispersed mycelium suitable for studying the physiological control of secondary metabolism also supported the formation of 5-hydroxy-4-oxonorvaline by S. akiyoshiensis. Key words: Streptomyces akiyoshiensis, mycelial aggregation, growth conditions.

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Enhancement of 2,3-Butanediol Production byKlebsiella oxytocaPTCC 1402

Journal of Biomedicine and Biotechnology ◽

10.1155/2011/636170 ◽

2011 ◽

Vol 2011 ◽

pp. 1-7 ◽

Cited By ~ 11

Author(s):

Maesomeh Anvari ◽

Mohammad Reza Safari Motlagh

Keyword(s):

Acetic Acid ◽

Taguchi Method ◽

Submerged Culture ◽

Klebsiella Oxytoca ◽

Culture Conditions ◽

Inoculum Size ◽

Medium Composition ◽

Operating Parameters ◽

Optimal Operating ◽

Mixing Intensity

Optimal operating parameters of 2,3-Butanediol production usingKlebsiella oxytocaunder submerged culture conditions are determined by using Taguchi method. The effect of different factors including medium composition, pH, temperature, mixing intensity, and inoculum size on 2,3-butanediol production was analyzed using the Taguchi method in three levels. Based on these analyses the optimum concentrations of glucose, acetic acid, and succinic acid were found to be 6, 0.5, and 1.0 (% w/v), respectively. Furthermore, optimum values for temperature, inoculum size, pH, and the shaking speed were determined as 37°C, 8 (g/L), 6.1, and 150 rpm, respectively. The optimal combinations of factors obtained from the proposed DOE methodology was further validated by conducting fermentation experiments and the obtained results revealed an enhanced 2,3-Butanediol yield of 44%.

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Production and Characterization of a Novel Bioflocculant from Bacillus licheniformis

Applied and Environmental Microbiology ◽

10.1128/aem.02558-09 ◽

2010 ◽

Vol 76 (9) ◽

pp. 2778-2782 ◽

Cited By ~ 100

Author(s):

Yuyan Xiong ◽

Yuanpeng Wang ◽

Yi Yu ◽

Qingbiao Li ◽

Haitao Wang ◽

...

Keyword(s):

Bacillus Licheniformis ◽

16S Rrna Gene Sequencing ◽

Amino Sugar ◽

Culture Conditions ◽

Inoculum Size ◽

Infrared Spectrometry ◽

Rrna Gene ◽

Neutral Sugar ◽

Sem Images ◽

Average Mass

ABSTRACT A bacterium producing an extracellular bioflocculant was isolated from contaminated LB medium and identified as Bacillus licheniformis by 16S rRNA gene sequencing and its biochemical/physiological characteristics. The optimum culture conditions for flocculant production were an initial medium pH of 7.2 and an inoculum size of 4% (vol/vol). The maximum flocculating activity (700 U/ml) was obtained after cultivation at 37°C for 48 h. Chemical analyses of the purified bioflocculant revealed that it was a proteoglycan composed of 89% carbohydrate and 11% protein (wt/wt). The mass ratio of neutral sugar, amino sugar, and uronic acid was measured at 7.9:4:1. Infrared spectrometry further indicated the presence of carboxyl, hydroxyl, and amino groups, typical of heteropolysaccharide. The average mass of the bioflocculant was calculated to be 1.76 × 106 Da. Scanning electron microscopy (SEM) images of the bioflocculant showed an irregular structure with netted texture. Its efficient flocculation capabilities suggest potential applications in industry.

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Design of a culture medium for optimal growth of the bacterium Pseudoxanthomonas indica H32 allowing its production as biopesticide and biofertilizer

AMB Express ◽

10.1186/s13568-020-01127-y ◽

2020 ◽

Vol 10 (1) ◽

Author(s):

Dayana Morales-Borrell ◽

Nemecio González-Fernández ◽

Néstor Mora-González ◽

Carlos Pérez-Heredia ◽

Ana Campal-Espinosa ◽

...

Keyword(s):

Hydrogen Sulfide ◽

Yeast Extract ◽

Culture Medium ◽

Nitrogen Sources ◽

Optimal Growth ◽

Culture Conditions ◽

Medium Composition ◽

Mineral Salts ◽

Molar Ratios ◽

Biotechnological Processes

Abstract Culture medium composition is one of the most important parameters to analyze in biotechnological processes with industrial purposes. The aim of this study was to design of a culture medium for optimal growth of the bacterium Pseudoxanthomonas indica H32 allowing its production as biopesticide and biofertilizer. The influence of several carbon and nitrogen sources and their molar ratios on P. indica H32 growth was investigated. The effect of different micronutrients such as mineral salts and vitamin on P. indica H32 growth was determined as well. A mixture design based on Design-Expert 10.0 Software was performed to optimize the culture medium concentration. Finally, in the designed medium, an attribute of the biological mechanism of action of the P. indica H32 against nematodes, was evaluated: the hydrogen sulfide production. It was found that tested carbon/nitrogen ratios were not a significant influence on P. indica H32 growth. Growth of P. indica H32 was favored with use of sucrose, yeast extract and phosphate buffer without the addition of any tested micronutrients. An optimal concentration of 10 g/L sucrose and 5 g/L yeast extract were obtained at a cost of 0.10 $/L. In this concentration, the specific growth rate (µ) and maximal optical density (Xmax) were equal to 0.439 h− 1 and 8.00 respectively. It was evidenced that under the culture conditions used, P. indica H32 produced hydrogen sulfide. The designed medium led to a 1.08 $/L reduction of costs in comparison to LB medium. These results were critical to carry on with biotechnological development of P. indica H32 as a bioproduct.

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Screening of Culture Conditions for Production of Xylanase from Landfill Soil Bacteria

Indonesian Journal of Chemistry ◽

10.22146/ijc.39709 ◽

2019 ◽

Vol 19 (2) ◽

pp. 470 ◽

Cited By ~ 2

Author(s):

Siti Nor Amira Rosli ◽

Rohaida Che Man ◽

Nasratun Masngut

Keyword(s):

Carbon Source ◽

Moisture Content ◽

Nitrogen Source ◽

Incubation Period ◽

Xylanase Activity ◽

Nitrogen Sources ◽

Culture Conditions ◽

Inoculum Size ◽

Xylanase Production ◽

Initial Ph

Culture conditions including initial pH media, incubation period, inoculum size, type of carbon source, type of nitrogen source and its concentration, which affect xylanase production were screened via the one-factor-at-a-time approach. The bacteria used in the production of xylanase was isolated from the landfill site at Sg. Ikan, Kuala Terengganu, Malaysia. Three characterizations of the landfill soil were investigated for their moisture content, ash content, and pH. The culture conditions range used in the experimental work were between 6–30 h for the incubation period, with initial pH between 5–9, inoculum size between 1–20% v/v, carbon, nitrogen sources, and nitrogen source concentration between 1–5% w/v. Xylanase activity was estimated using dinitrosalicylic acid (DNS) based on the release of xylose under standard assay conditions. The landfill soil was observed to have pH between pH 3.4–7.2 with a moisture content between 12.4–33.7% and ash ranged between 3.5–4.3%. Results showed that the highest xylanase activity within studied ranges was recorded at 25.91±0.0641 U/mL with 10% (v/v) inoculum size, 1% (w/v) xylose as sole carbon source, mixture of 1% (w/v) peptone and 0.25% (w/v) ammonium sulphate as nitrogen sources, which was carried out at initial pH of 8.0 for 24 h incubation.

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Production of a Bioflocculant by Penicillium Sp. HHE-P7 Using Sugarcane Molasses

Advanced Materials Research ◽

10.4028/www.scientific.net/amr.518-523.453 ◽

2012 ◽

Vol 518-523 ◽

pp. 453-459

Author(s):

Li Fan Liu

Keyword(s):

Rotation Speed ◽

Fermentation Broth ◽

Nitrogen Sources ◽

Culture Conditions ◽

Inoculum Size ◽

Carbon And Nitrogen Sources ◽

Carbon And Nitrogen ◽

Medium Components ◽

Flocculating Activity ◽

Initial Ph

Bioflocculant MBF7 was produced by a novel bioflocculant-producing microorganism HHE-P7. In order to reduce the bioflocculant producing cost, culture experiments were conducted. The effects of medium components including carbon and nitrogen sources as well as culture conditions such as pH of molasses diluents, cultivating temperature, inoculum size were investigated. The results showed when the molasses waste was diluted at COD concentration of 2000 mg/L, the optimal culture conditions for MBF7 production by HHE-P7 were inoculum size 1% (v/v), initial pH 5, cultivating temperature 25°C at the rotation speed 150 r/min. Under such conditions, MBF7 had a flocculating activity of 83% for 5 g/L kaolin clay suspension. About 3.19 g crude bioflocculant could be recovered from 1.0 L of molasses fermentation broth.

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Random mutagenesis and process optimization of bacterial co-culture for hyperproduction of 1,4-α-D-glucan glucanohydrolase using submerged fermentation

Hemijska industrija ◽

10.2298/hemind180213022a ◽

2018 ◽

Vol 72 (6) ◽

pp. 329-339

Author(s):

Roheena Abdullah ◽

Samra Kiran ◽

Mehwish Iqtedar ◽

Afshan Kaleem ◽

Faiza Saleem ◽

...

Keyword(s):

Uv Light ◽

Carbon Sources ◽

Random Mutagenesis ◽

Strain Improvement ◽

Nitrogen Sources ◽

Tween 80 ◽

Fold Increase ◽

Inoculum Size ◽

Medium Composition ◽

Chemical Mutagenesis

The exponential increase in the application of 1,4-?-D-glucan glucanohydrolase (GGH) in various fields has placed stress and demand in both qualitative improvement and quantitative enhancement through strain improvement. In the present work, Bacillus subtilis LCBT-15 and Bacillus amyloliquefaciens LCBT-20 were subjected to physical as well as chemical mutagenesis for improving the GGH production potential. Applications of the UV light and ethidium bromide did not cause a significant increase in the enzyme production. However, Ethyl methane sulphonate (EMS) treated co-culture 10 gave 1.3-fold increase in the GGH production, in contrast to the wild co-culture. Different physicochemical parameters including fermentation media, rate of fermentation, temperature, pH, nitrogen and carbon sources and surfactants were also investigated. The M7 medium composition was optimized for GGH production after 48h of incubation at 37?C and pH 6. The optimum inoculum size was 3.5 ml (1x106 cells/ml) in 50 ml of medium. The best carbon and nitrogen sources were lactose (2.5 %); ammonium chloride (1.75 %) and beef extract (1 %), respectively. Optimal GGH production (287 U/ml) was obtained when the medium was supplemented with 0.05% Tween 80. The novelty of this work was exploration of the synergistic phenomena of mutant bacterial co-culture for the enhancement of GGH production.

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Classical and Statistical Optimization of Medium Composition for Promoting Prodigiosin Produced by Local Isolate of Serratia Marcescens

Al-Khwarizmi Engineering Journal ◽

10.22153/kej.2018.03.006 ◽

2018 ◽

Vol 14 (4) ◽

pp. 92-102 ◽

Cited By ~ 2

Author(s):

Sura Jasem Mohammed ◽

Khalid Jaber Kadhum ◽

Khalid Waleed Hameed

Keyword(s):

Serratia Marcescens ◽

Trace Element ◽

Nitrogen Sources ◽

Antimicrobial Activities ◽

Inoculum Size ◽

Medium Composition ◽

Statistical Optimization ◽

Statistical Experimental Design ◽

Carbon And Nitrogen ◽

Anticancer Properties

Prodigiosin is a ‘natural red pigment produced by Serratia marcescens which exhibits immunosuppressive and anticancer properties in addition to antimicrobial activities. This work presents an attempt to maximize the production of prodigiosin by two different strategies: one factor at time (OFAT) and statistical optimization. The result of OFAT revealed that sucrose and peptone were the best carbon and nitrogen sources for pigment production with concentration of prodigiosin of about 135 mg/ L. This value was increased to 331.6mg/ L with an optimized ratio of C/N (60:40) and reached 356.8 with pH 6 and 2% inoculum size at end of classical optimization. Statistical experimental design based on Response surface methodology was conducted to optimize the composition of trace element. The design revealed that the predicted prodigiosin production of 406 mg/L can be achieved when concentrations of trace element CaCl2·2H2O, FeSO4·4H2O, MgSO4·7H2O and MnSO4·4H2O were equal to 9.22, 0.32, 0.67 and 2.48 g/L, respectively. The actual production of prodigiosin in the optimized medium was 375 mg/L. Growth kinetics of S. marcescens were evaluated in optimized medium which revealed that prodigiosin was ‘non-associated growth’ secondary metabolite with maximum production of approximately 365.7 mg/L obtained after 54 hours of incubation.

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Fibrinolytic enzymes from a newly isolated marine bacteriumBacillus subtilisA26: characterization and statistical media optimization

Canadian Journal of Microbiology ◽

10.1139/w09-057 ◽

2009 ◽

Vol 55 (9) ◽

pp. 1049-1061 ◽

Cited By ~ 34

Author(s):

Rym Agrebi ◽

Anissa Haddar ◽

Mohamed Hajji ◽

Fakher Frikha ◽

Laila Manni ◽

...

Keyword(s):

Enzyme Production ◽

Methodological Approach ◽

Statistical Design ◽

Maximal Activity ◽

Culture Conditions ◽

Medium Composition ◽

Carbon Substrate ◽

Rrna Gene ◽

Fibrinolytic Enzymes ◽

Initial Medium

A fibrinolytic enzyme producing bacterium was isolated and identified as Bacillus subtilis A26 on the basis of the 16S rRNA gene sequence. The fibrin zymography analysis reveals the presence of at least three fibrinolytic enzymes. The crude enzyme exhibited maximal activity at 60 °C and pH 8.0. Medium composition and culture conditions for the enzyme production by B. subtilis A26 were optimized using two statistical methods. The Plackett–Burman statistical design was applied to find the key ingredients and conditions for the best yield of enzyme production. Five significant variables (hulled grain of wheat, casein peptone, NaCl, CaCl2, and initial pH) were selected for the optimization studies. The response surface methodological approach was used to determine the optimal concentrations and conditions. The optimized medium contained 40.0 g·L–1hulled grain of wheat, 3.53 g·L–1casein peptone, 4.0 g·L–1CaCl2, 3.99 g·L–1NaCl, 0.01 g·L–1MgSO4, and 0.01 g·L–1KH2PO4, pH 7.78. The medium optimization resulted in a 4.2-fold increased level of fibrinolytic production (269.36 U·mL–1) compared with that obtained with the initial medium (63.45 U·mL–1). A successful and significant improvement in the production of protease by the A26 strain was accomplished using inexpensive carbon substrate (hulled grain of wheat), allowing a significant reduction in the cost of medium constituents.

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