Open Access Original Research Article

Extraction and Purification of Protease from Nigella sativa for Its Potential Use in Celiac Disease

Gamal A. Gabr

Asian Journal of Biotechnology and Bioresource Technology, Page 1-9
DOI: 10.9734/AJB2T/2018/44555

Celiac disease (CD) is a disease of the digestive system resulting from the intolerance of autoimmune system against gluten and gliadin proteins in wheat, barley, rye and some varieties of oats. In this work protease was isolated and partially purified from Nigella Sativa using ammonium sulphate, acetone and trichloroacetic acid. The purified protease was evaluated further for its possible use in celiac disease by its ability to hydrolyse both, gluten and gliadin. Data indicate that the highest purity of digestive protease enzymes was obtained by acetone 80% and 0.2M trichloroacetic acid (TCA). The enzyme purified by acetone exhibited the high value of Vmax (166.66 mg/min) and the low value of Km (0.133 mg/ml). When the obtained enzyme was tested to hydrolyse gluten and gliadin at two increasing concentrations of 1 and 2 mg/ml, almost similar efficacy was observed (28.46% and 32.02%; for gluten) and (22.7% and 16.84%; for gliadin), whereas Digestin, a positive control which contain Papain and Sanzyme 3500 and works by helping in the breakdown of proteins, produced higher efficacy at 2 mg/ml to hydrolyse gluten (38.67%) with 1 mg/ml (25.68%) whereas similar results were observed against gliadin at both concentrations (22.06% and 22.5%). This study show the efficacy of protease obtained from Nigella Sativa as a natural and economical source for the hydrolysis of gluten and gliadin, which is considered the primary reason for celiac disease.

Open Access Original Research Article

Enhancement of Triterpenoids formation of Antrodia cinnamomea in thin Stillage-submerged Culture by Shifting the Shaking Rate and Adding Soybean Oil

Te-Wei Ma, Min-Yu Chiu, Fan-Chiang Yang

Asian Journal of Biotechnology and Bioresource Technology, Page 1-13
DOI: 10.9734/AJB2T/2018/44690

Antrodia cinnamomea is an endemic medical mushroom in Taiwan. It is well known that the major effective components in this medical fungus are polysaccharides, triterpenoids, and steroids; it is found only in Taiwan. Its anti-tumour activity remains Current research interest is focused on its anti-tumour activity from secondary metabolites of triterpenoids. However, the main disadvantage of mycelia produced by liquid cultures is the quite small amount of triterpenoid. In this study, various cultivation strategies were tested to investigate their effects on the production of triterpenoids in thin stillage submerged cultures of A. cinnamomea. The results indicate that when thin stillage was reused in shaking flask culture at a constant speed, crude triterpenoids production reached the level of 43.04 mg/L at a low inoculum size of 1% at d-20, which was twice that of the control of high inoculum size of 10%. This study demonstrates that diminishing inoculum size has a significant effect on mycelia morphology, and could raise the generation of crude triterpenoids. The addition of soybean oil and corn starch could improve the production of crude triterpenoids to 78.53 mg/L, triple that of the control. Shifting the shaking rate was the most efficient method for improving triterpenoids production. When the shaking rate was shifted at d-8 and the rate maintained for 5 d, the level of crude triterpenoids rose to 152.62 mg/L, which was 7-fold that of the control. The results also demonstrate the feasibility of the reuse of thin stillage for liquid cultures of A. cinnamomea.

Open Access Original Research Article

Retracted: Bioactive Compounds Identified from Cellulytic Bacteria Isolated from Food Dissipated and Their Screening for Potential Antimicrobial and Cytotoxicity Activities

M. R. Mahmud, Sajib Podder, Rima .

Asian Journal of Biotechnology and Bioresource Technology, Page 1-9
DOI: 10.9734/AJB2T/2018/43320

Food waste is a rich source of lingo-cellulosic materials thus providing an intense environment for the growth of cellulolytic bacteria. The present study was conducted to isolate and identify bioactive compound of cellulose degrading bacteria from food waste and their screening for potential antimicrobial activity. Cellulytic bacteria Klebsiellia pneumoniaee strain ZJ-02 identified was isolated from food waste. To test the sensitivity of the isolates, five different antibiotics were used. The strains were tested for resistance to Neomycin 30µg, Ampicillin 10mg, Cephalosporin 30µg, Doxycycline 30µg, and Erythromycin 15µg. When tested by disc diffusion method on antibiotic spreaded nutrient agar plate the strains showed sensitivity to Neomycin, Ampicillin, Cephalosporin, Doxycycline, and Erythromycin. The ethyl acetate extract of Klebsiellia pneumoniae strain ZJ-02   showed cytoxicity against brine shrimp nauplii with the LD50 value of 50.486 µg/ml and showed potent anti-bacterial activity against five human pathogenic bacteria Listeria monocytogenes, Bacillus subtilis, Shigella boydii, Escherichia coli, Klebsiella pneumoniae. Minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) have been determined. The MIC and MBC values were between 32-64μg/ml and 64-128μg/ml. Higher MBC value than that of MIC indicates Klebsiellia pneumoniae strain ZJ-02 is bacteriostatic. In addition, potential active compounds were detected using gas chromatography and mass spectrometry- 1,2-benzenedicarboxylicacid, Pyrrolo[1,2-a]pyrazine-1,4-dione, 3,6-Diisobutyl 2,5piperazinedion, trimethylsilyl 3,5-dimethoxy-4-(trimethylsilyloxy) benzoate, 3H-cycloocta[c]pyran-3-one and phthalic acid were found in EAEKP. In summary, our findings provide insight into the Klebsiellia pneumoniae strain ZJ-02 from food waste are a potential source of novel antibiotic leads that are worthy of further study.


Retraction Notice: This paper has been retracted from the journal. This journal is determined to promote integrity in research publication. This retraction is in spirit of the same. After formal procedures editor(s) and publisher have retracted this paper on 25th November-2019. Related policy is available here:

Open Access Original Research Article

Induction of Anthocyanin Production in Established Callus Cultures of Roselle (Hibiscus sabdariffa L.) Using Yeast Extract

Therese Julienne T. Medina, Lourdes B. Cardenas

Asian Journal of Biotechnology and Bioresource Technology, Page 1-8
DOI: 10.9734/AJB2T/2018/45918

Aims: Hibiscus sabdariffa var. sabdariffa (Malvaceae), a shrub locally known as roselle has been valued for its vibrant red colored calyces that are used as food colorant and health drink. Its anthocyanin content has been known to have health promoting effects like antioxidant activity, antimicrobial and anti-cancer, among others. This study was done to establish callus cultures of roselle which are capable of producing anthocyanins.

Study Design: An experimental study was done to look into the effects of different factors such as explant source and growth hormone concentration on the induction of roselle callus cultures. The effect of different concentrations of yeast extract as elicitor of anthocyanin production was also tested.

Place and Duration of Study: The study was conducted at the Biotechnology Laboratory for Natural Products, Institute of Biological Sciences, University of the Philippines Los Baños from June 2011 to May 2014.

Methodology: Callus induction was done using aseptically grown seedlings of the Thailand accession at MS medium with different combinations of growth hormones (T1: 0.5ppm 2,4-D and 1ppm kinetin; T2: 1ppm 2,4-D and 1ppm kinetin; T3: 1ppm 2,4-D and 2ppm kinetin). Established cultures were subjected to anthocyanins elicitation using yeast extract (Y1: 8g/L, Y2: 4g/L, Y3: 1g/L and Yc: no extract) as a biotic elicitor.

Results: Callus formation and ephemeral anthocyanin production were observed 2 weeks after inoculation. Addition of yeast extract increased the growth rate up to 10-fold (4g/L) but difference among treatments was not statistically significant. Callus cultures produced anthocyanins 2 weeks after transferred back to a growth medium without yeast extract.

Conclusion:  Anthocyanin production was unstable and temporary but the calli proved competent for anthocyanin production. Yellow calli were also observed after exposure to yeast extract, TLC profile showed presence of chlorogenic acids which are possible precursors for anthocyanin production.

Open Access Original Research Article

The Production of Liquid Biofertilizer from Cassava Peels and Spent Mushroom Substrates Using Microbial Inoculants

O. C. Okolie, H. O. Stanley, Frank-Peterside, N., C. J. Ugboma

Asian Journal of Biotechnology and Bioresource Technology, Page 1-13
DOI: 10.9734/AJB2T/2018/45642

This study showed the production of liquid biofertilizer from organic wastes (cassava peels and spent mushroom substrates) using microbial inoculants (Bacillus spp., Pseudomonas spp., Lactobacillus spp. and Aspergillus spp.). The organic wastes were autoclaved and then immersed in sterile water inside plastic pots. The plastic pots were twelve in number (labelled A to L) with duplicates but one of it (L) serves as the negative control (sterile water + organic wastes). They were used as the composting reactors with different compositions of microbial inoculants. There were increase in the physicochemical parameters of the liquid compost in week one and two respectively. Generally there was slight decrease in the microbial population at week two. On the addition of Azotobacter spp. the microbial population increased substantially along with some physicochemical parameters (nitrates, carbon(iv)oxide, total organic carbon, potassium and phosphates), only the pH and Total Nitrogen concentrations showed  reduction in their concentrations. All compost reactors were then mixed together and filtered to obtain the liquid biofertilizer. The shelf life of the liquid biofertilizer was determined at monthly intervals from December to April. The highest microbial population was observed in the third week of February (3.6 x 108 cfu/l). The efficacy of liquid biofertilizer produced were tested on the growth index of plant (Maize and Okra) and compared with a commercial organic liquid fertilizer and a control (no fertilizer application). The number of leaves, size of stem girth and the height of plant in maize and okra favoured the liquid biofertilizer applications compared to the commercial Organic liquid fertilizer. To determine the most efficient mode of fertilizer application, foliar applications and soil inoculation were tested on the crops using the liquid biofertilizer and the commercial organic liquid fertilizer until week five. It was discovered from this research that soil inoculation is the best method for the application of the liquid biofertilizer.