Open Access Short Research Article

Extraction and Characterization of Chitosan Obtained from Scales of Clarias gariepinus (Catfish)

E. Gokulalakshmi, K. Ramalingam, Umasankari ., M. C. Vanitha

Biotechnology Journal International, Page 1-8
DOI: 10.9734/BJI/2017/27944

Fish waste is considered to be one of the major bio-pollutants which are generally discarded in coastal regions through local markets and fish processing industries. Nowadays, it is being eyed as a newer bioresource. Fish scales are good source of chitin and chitosan. Very little information is available related to extraction of chitosan from fish scales. In this study fish scale waste from markets around Chennai was used for extraction of chitosan. Chitosan is extracted from the fish scales and the physicochemical properties of the extracted chitosan were characterized by FTIR, UV-Visible spectroscopy, and Scanning Electron Microscopy. The antibacterial activity of chitosan against different sea food pathogens pathogens like Vibrio parahaemolyticus, Vibrio cholerae, Staphylococcus aureus, Salmonella typhii, Escherichia coli and Shigella dysenteriae was evaluated by calculation of minimum inhibitory concentration (MIC). Antioxidant activity for chitosan was performed with various concentration of 20 µm-100 µm and the scavenging activity of the sample is calculated as 15%. The physiochemical properties and FTIR and XRD studies confirm the production of chitosan, because of its high antibacterial activity against pathogens, hence chitosan can be widely used in food preservation, manufacture of wound dressing and in antimicrobial finished textiles.

Open Access Original Research Article

Production of Monoclonal Specific Antibody against Brazzein Fusion Protein

Fatemeh Mansouri, Mohammad Hossain Modarressi

Biotechnology Journal International, Page 1-9
DOI: 10.9734/BJI/2017/33505

Background: The use of a natural sweet protein is important as an attractive alternative to sucrose in the human diet. Rigorous research has identified the use of the artificial sweetener that leads to various diseases, cancer and heart disease. Many functional studies are necessary for examining the structure and biological activity of natural sweeteners such as Brazzein and its receptor targeting. In this report, we produced a panel of monoclonal antibodies against the recombinant brazzein which can be used for functional studies.

Methods: Hybridomas cell fusion technology was used for the production of monoclonal antibody. BALB/c mice were immunized twice with purified fusion proteins with Freund adjuvant. Next, 14 days after the last injection, blood sample was collected from the eye and serum was separated and the antibody titer was detected by enzyme-linked immunosorbent assay (ELISA). The mouse with the highest antibody titers was selected. Seven days before fusion, the selected mouse was boosted with 50 µg antigen into the peritoneum to develop a robust immune response and then spleen cells were isolated from the same animal for fusion to myeloma SP2/0 to generate hybridoma cells.

Results: Hybridoma clones were screened by ELISA and their monoclonal antibodies were purified. Specificity and reactivity of the monoclonal antibodies were determined by western blot analysis. We produced a panel of monoclonal antibodies that were highly specific and reacted with Brazzein protein that was detected by western blot analysis.

Conclusion: These monoclonal antibodies bind to the MBP (Maltose binding protein)-containing recombinant protein in western blotting, so this antibody can be a useful probe for tracing proteins in basic research and pharmaceutical applications.

Open Access Original Research Article

Isolation and Characterization of Bacteriocinogenic Enterococcal and Lactococcal Strains from South of Morocco Dairy Product

Abdelkhaleq Elmoslih, Mariem Zanzan, Rabha Aissa, Fatima Hamadi, Ghita Ait Baddi, Abdellah Ait Ben Aoumar, Fouad Achemchem

Biotechnology Journal International, Page 1-16
DOI: 10.9734/BJI/2017/32919

Aim: To investigate the occurrence of bacteriocinogenic lactic acid bacteria (BAL) in different animal’s milk of the south of Morocco.

Place and Duration of Study: Laboratory of Microbial biotechnologies and plant Protection, Faculty of Sciences, and Bioprocess and Environment laboratory (LASIME), EST-Agadir, Ibnou Zohr University, Agadir, Morocco, between January 2014 and January 2016.

Methodology: A total of 2000 different colonies, isolated from 42 samples of dromedary, ewe’s, goat and cow spontaneously fermented milk collected from some southern regions of Morocco, were tested for antimicrobial activity. Three indicator strains were used; Listeria innocua, Bacillus subtilis and Enterococcus hirae. The selected strains are phenotypically and biochemically identified, especially by API 20 Strep galleries. In addition, the sanitary and technological aspects of these strains are studied.

Results: Among the active strains 150 strains were selected, and 91% among them were identified as lactic acid bacteria. Out of these, 11 strains isolated from dromedary and ewe’s milk are shown to be active by the agar well diffusion assay (AWDA). Seven (7) strains were identified as Enterococcus faecium, three (3) as Enterococcus faecalis and only one (1) strain was identified as Lactococcus lactis. The twelve strains are active against a wide range of pathogenic and spoilage bacteria, including Listeria monocytogenes, Staphylococcus aureus and Escherichia coli. In addition, all of these strains shown to lack haemolytic, decarboxylatic, proteolytic and lipolytic activities and to be susceptible to most tested antibiotics.

Conclusion: These results suggest a potential application of isolated strains of lactic acid bacteria in bio-preservation of fermented foods especially dairy products.

Open Access Original Research Article

Enhancing the Performance of Three White-rot Fungi in the Mycoremediation of Crude Oil Contaminated Soil

Moses B. Adewole, Olumuyiwa O. Olanrewaju

Biotechnology Journal International, Page 1-10
DOI: 10.9734/BJI/2017/34267

Contamination of soils by petroleum hydrocarbon is on the increase, particularly in the oil producing areas of Nigeria. White-rot fungi have enzymes which are capable of turning these organic compounds into harmless substances. This work investigated the performance of three white-rot fungi (Pleurotus tuber-regium, P. ostreatus and P. pulmonarius) for the remediation of different concentrations (0, 1, 2.5, 5, 10 and 20% w/v) of crude oil contaminated soils with sawdust and poultry manure as bedding materials. Ten grammes of each of the white-rot fungi were separately inoculated in each bottle containing exhaustively cropped topsoil (200 g), rice straw (40 g) and wheat bran (20 g). Each treatment was conducted in three replicates and arranged in a 3 x 6 x 3 complete randomised design. The bottles (54) were incubated for zero, one and three months in a dark room, exposed to light, watered daily thereafter for twelve days for fruiting bodies to spring out and harvested. P. ostreatus had best agronomic performance and P. tuber-regium removed hydrocarbons and heavy metals more than either P. ostreatus or P. pulmonarius under similar experimental conditions. The yield and mycoremedial performance of the three tested white-rot fungi demonstrated potentials for cleaning-up petroleum hydrocarbon contaminated soil, but their performance reduced from 5.0% (w/v) crude oil substrate contamination.

Open Access Review Article

Diabetes Mellitus: Can Stem Cells be the Answer?

M. Senthilnathan, A. Ramadevi, K. x K. Srinivas, A. Thangamani

Biotechnology Journal International, Page 1-12
DOI: 10.9734/BJI/2017/34614

This review aims to enlighten the readers regarding the past, present and future of stem cells in the treatment of Diabetes. Diabetes is one of the leading causes of morbidity and mortality, affecting more than 415 million people worldwide.  It is estimated that one in ten adults will have diabetes by 2030. Diabetes is mainly due to reduction in β-cell mass which are responsible for insulin production. Exogenous administration of insulin is having good impact on restoring glucose homeostasis, but it does not entirely control the minute-to-minute fluctuations in systemic blood glucose. Recently cellular-based therapies have been established for exogenous insulin administration by modern pump technology. One of the most interesting therapies involves substitution of insulin producing islet cells by transplantation. But lack of donor material and lifelong immunosuppression made the technique unfeasible. These restrictions have led to exploration of other sources of β-cells, one of the prospects being the stem cells. Several types of stem cells have been used to make pancreatic β-cells, including human embryonic stem cells / induced pluripotent stem cells, pancreatic stem / progenitor cells, and non-pancreatic stem cells. There is also evidence of adult β-cells regeneration through β-cell replication and cellular reprogramming. Functional restoration of existing β-cells, transplantation of stem cells or stem cell-derived β-like cells might provide new opportunities for treatment. In conclusion it can be said that the research is still wide open to arrive at the efficient reprogramming of various types of stem cells to destine them towards functional β-cells.