Open Access Original Research Article

Cell Proliferation Using Broccoli Leaf Cutting in vitro Culture: Its Biochemical and Antioxidant Properties

A. B. M. Sharif Hossain, Imdadul Haq, Nasir Adam Ibrahim, Mohammed Aleissa

Biotechnology Journal International, Page 1-9
DOI: 10.9734/BBJ/2016/22322

Significance of the study: Tissue or cell culture keeps a significant role in micro-propagation in the plant production industry. Nowadays it has been successfully performed from many species in plant. Millions of explants can be produced by tissue or cell culture per year in any plant production industry. 

Aim: The study was conducted to investigate the root, shoot and leaf formation from the leaf cutting in vitro culture of broccoli (Brassica oleracea var italic).

Methodology: Different BAP (6-Benzylaminopurine) and IBA (Indole-butaric-acid) hormone (0.25, 0.50, 1.0, 1.50, 2.0 2.5, 3.0 and 3.5 mg/l) concentrations in combination with MS media was used to culture leaf cutting in vitro.

Results: The results showed that there was no shoot formed, but a few leaves and bulky callus proliferation were appeared. The highest number (6.75) of root proliferation was found in the concentration of 2.0 mg/l IBA + 0.25 mg/l BAP combination. 2,2-diphenyl-2-picrylhydarzyl (DPPH) free radical scavenging potential was higher (70%) in leaves extract than in callus extracts (46%) at concentration of 10 mg/ml.

Conclusion: The present results conclude that it is better to use the combination of BAP and IBA at different concentration to produce root proliferation, leaf initiation and callus formation in broccoli.

Open Access Original Research Article

Dietary Consumption of Citrullus lanatus can Ameliorate Infertility Potential of Carica papaya Seeds Extract in Male Rats

Nathaniel Ohiemi Amedu, Ukanu Paul Idoko

Biotechnology Journal International, Page 1-9
DOI: 10.9734/BBJ/2016/22805

Aim: This study examined dietary consumption of Citrullus lanatus as an ameliorative agent against infertility potential of seeds extract of Carica papaya in male wistar rats.

Study Design: Eighteen adult male Wistar rats were randomly assigned into three groups of six (n=6): A, B, and C. Group A served as the control group while the other groups served as the treated groups.

Place and Duration of Study: This study was carried out in the Animal Holdings of the Department of Anatomy, University of Ilorin, Ilorin, between July, 2014 and October, 2014.

Methodology: The Group A animals were given feed and water liberally throughout the study. Group B received 150 mgkg/bwt/d of Carica papaya extract intraperitoneally for 8 weeks, Group C received 150 mgkg/bwt/d of Carica papaya extract intraperitoneally plus Citrullus lanatus orally for 8 weeks. The rats were sacrificed at the end of administration, Testosterone, FSH and LH levels were assayed. Also, semen analysis was carried out to ascertain the level of other parameters.

Results: Generally there was significant reduction in the level of Testosterone, FSH and LH in Group B when compared with the Control and Group C. Also, there was significant reduction in the level of sperm count, motility, concentration and morphology in Group B when compared with the Control and Group C. However, there was no significant difference when almost all these parameters were directly compared between Control and Group C.

Conclusion: This study has demonstrated that dietary consumption of Citrullus lanatus can ameliorate infertility potential of Carica papaya seeds extract in adult male Wistar rats.

Open Access Original Research Article

Biovalorization of Olive Mill Waste Water for the Production of Gellan Gum from Sphingomonas paucimobilis

Ioannis Giavasis, Konstantinos Petrotos

Biotechnology Journal International, Page 1-15
DOI: 10.9734/BBJ/2016/22510

Aims: The aim of this work was to investigate and optimize the potential of olive mill waste water (OMWW) to be utilized as a substrate for the production of gellan gum by Sphingomonas paucimobilis.

Study Design: The study was divided in two phases, a preparatory phase for the removal of olive polyphenols and/or condensation of OMWW, and a bioprocessing-fermentation phase.

Place and Duration of Study: The study was conducted at the TEI of Thessaly from January 2013 to August 2015.

Methodology: OMWW was used as a fermentation substrate for production of gellan after dephenolization by microfiltration, thermal condensation, and addition of minerals/nitrogen sources and glycerol. S. paucimobilis was grown in this substrate under controlled process conditions in shake flasks and a 15 lt bioreactor. Biomass, gellan, sugars, phenol concentration and apparent viscosity of OMWW were evaluated.

Results: The results show that although S. paucimobilis can degrade olive polyphenols in OMWW, the removal of polyphenols is beneficial to gellan synthesis. The condensation (2/1, i.e. to the half of its initial volume) of the dephenolized OMWW also improved gellan production as it offered more sugars for polysaccharide production. After both dephenolization and 2/1 condensation gellan production increased by 50%. Yeast extract (1 g/l) was the preferable nitrogen source supplementation as it stimulated both cell growth and gellan synthesis. Glycerol (5 g/l) increased gellan formation and viscosity of the fermentation broth, which may indicate a key role of glycerol in gellan biosynthesis. An agitation of 500 rpm and aeration of 1 vvm resulted in the highest gellan production of 9.5 g/l in 63 h in the optimized and fortified OMWW after dephenolization and condensation. At 500 rpm an intense aeration of 2 vvm increased cell growth at the expense of gellan formation and resulted in a reduced viscosity. 

Conclusion: After dephenolization, condensation, addition of some nitrogen source and glycerol, OMWW can be utilized as a substrate for efficient gellan production.

Open Access Original Research Article

Biovalorization of Olive Mill Waste Water for the Production of Single Cell Protein from Saccharomyces cerevisiae, Candida utilis and Pleurotus ostreatus

Ioannis Giavasis, Konstantinos Petrotos

Biotechnology Journal International, Page 1-16
DOI: 10.9734/BBJ/2016/22509

Aims: The aim of this work was to investigate and optimize the potential of olive mill waste water (OMWW) to be utilized as a substrate for the production of single cell protein (SCP).

Study Design: The study was divided in two phases, a preparatory phase for the removal of olive polyphenols and/or condensation of OMWW, and a bioprocessing-fermentation phase.

Place and Duration of Study: The study was conducted at the TEI of Thessaly from January 2013 to August 2015.

Methodology: OMWW was used as a fermentation substrate for production of SCP after dephenolization by microfiltration, condensation via reverse osmosis, and addition of minerals/nitrogen sources. Saccharomyces cerevisiae (S. cerevisiae), Candida utilis (C. utilis) and Pleurotus ostreatus (P. ostreatus) were grown in this substrate under controlled process conditions in shake flasks and a 15 lt bioreactor. Biomass, sugars, phenol concentration of OMWW and the protein content of the harvested biomass were evaluated.

Results: Dephenolization of OMWW is essential for sufficient growth of both yeasts, while                      P. ostreatus grows better in the untreated substrate, as it can degrade the polyphenols while the dephenolization step reduced also the sugar content of OMWW. Optimum process conditions for    S. cerevisiae included a pH 7, 350 rpm agitation rate, addition of (NH4)2SO4 to a 3/1 condensed medium, while C. utilis grew best at pH 5, 250 rpm, after addition of NH4NO3 in a 3/1 condensed medium. Maximum biomass concentration was 13.5 g/l and 14.6 g/l, respectively. 14.8 g/l of             P. ostreatus biomass were obtained at pH 5, 150 rpm after addition of NH4NO3. This mold had the highest yield but lowest productivity due to slow growth.   

Conclusion: P. ostreatus is a good producer of SCP in OMWW and reduces its organic load, but it has lower protein content. C. utilis had the highest SCP productivity, and the two yeasts had the highest protein concentration but require dephenolization of OMWW.

Open Access Original Research Article

Genetic Improvement of Saccharomyces boulardii R7 and Generate Suitable Strains for Synthesis and Expression of Recombinant Products

Majid Hussein Al-Jailawi, Amer Al-Shekdhaher, Rahem Al-Zaiadi

Biotechnology Journal International, Page 1-9
DOI: 10.9734/BBJ/2016/22927

Saccharomyces boulardii R7 was mutagenized using UV radiation. Uracil auxotroph mutants were isolated and four of these mutants showed stable mutation. Two mutants SbR7M7 and SbR7M10 were chose for transformation according to their similar behavior with wild type (SbR7) isolate. The two mutants (SbR7M7 and SbR7M10) were transformed with pYES2, which was extracted from Escherichia coli Top10, and two transformants (SbR7T7, SbR7T10) obtained. The transformation was confirmed by isolating the plasmid (pYES2) from these transformants and used to transform E.  coli Top10 (free of plasmid). Some probiotic properties were studied for the two transformants (SbR7T7, SbR7T10) compared with SbR7. They showed a noticed improvement in autoaggregation ability, an improvement of antagonistic activity toward E. coli: O157:H7 and Candida albicans and they reduced cholesterol ratio after 24 hr of incubation, however, the ratio increased after 48 hr of incubation.