Open Access Original Research Article

Partially Purified 3-Mercaptopyruvate Sulphurtransferase Obtained from the Intestine of Cane Rat (Thryonomys swinderianus) as a Detoxifier of Cyanide

A. A. Sanni, O. M. Ige, G. B. Olagunju, B. A. Olukade

Asian Journal of Biochemistry, Genetics and Molecular Biology, Page 1-9
DOI: 10.9734/ajbgmb/2020/v6i230146

Cane rat (Thryonomys swinderianus) is an herbivorous animal which feeds on plant materials, including cassavas that are known to contain cyanogenic glycosides. Cyanide or cyanogenic glycosides are known to be toxic for animal consumption. Therefore, Cane rat must have an inherent mechanism for detoxifying cyanide to be able to survive on its food. Previous works on 3MST have been done on several tissues of cane rats other than the intestine. In this work, we characterized and explored a partially purified 3MST (cyanide detoxifying enzyme) from the intestine of a cane rat for a possible therapeutic source against cyanide poisoning in other mammals that are susceptible to the toxin. 3MST from the intestine of T. swinderianus had a yield of 10.3% with specific activity of 0.21Umg-. The Km and Vmax values of the 3-MST were determined to be 40 mm and 0.20µmol/ml/min respectively for KCN (Potassium Cyanide); also 33.3 mm and 0.15 µmol/ml/min for mercaptoethanol. 3-MST presents in the intestine of T. swinderianus plays a significant role in detoxification of cyanogenic compounds, which makes it an effective target for cyanide poisoning therapy.

Open Access Original Research Article

Comparative Study of Ethanolic Wild African Nutmeg (Pycnanthus angolensis (Welw.) Stem Bark Extract Potentials and Selected Conventional Toothpaste against Hidden Resident Mouth Cavity Microfora

mouth cavity microfora., Teniola Temitayo Mary

Asian Journal of Biochemistry, Genetics and Molecular Biology, Page 10-36
DOI: 10.9734/ajbgmb/2020/v6i230147

The aim of the study is to evaluate and compare the antibacterial activity of ethanolic stem extract of (Wild African nutmeg) Pycnanthus angolensis (Welw.) and some commercially available toothpaste against bacteria isolated from the hidden resident mouth cavity microfora. Bacteria were isolated from swabs of apparently healthy individuals and were identified using Staining procedure biochemical tests and the use of Bergey’s manual of bacteria identification  The assay for antibacterial activity of Pycnanthus angolensis stem bark extract and the four toothpastes were determined using agar well diffusion method. The Gram positive bacteria isolated were Streptococcus sangus, Streptococcus ratti, Stomatococcus mucilaginous., Peptostreptococcus  sp., and Streptococcus mutans and the Gram negative bacteria were Veillonella atypical, Veillonella parvula, Veillonella dispar and Acidiaminococcus sp. Oral B toothpaste showed maximum efficacy of inhibition with inhibition zone diameter as wide as 20 mm at 100 mg/ml. Percentage frequency distribution of antibacterial activity of conventional toothpaste (Close-up) against hidden resident mouth cavity microfora depicts Acidaminococcus sp.13%, Veillonella parvula (10%), Veillonella dispar (12%), Peptostreptococcus  sp.(12%), Stomatococcus mucilaginous.(9%), Streptococcus ratti (13%), Veillonella atypical (11%), Streptococcus sangus (9%) and Streptococcus mutans (11%), Percentage frequency distribution of antibacterial activity of conventional toothpaste (Oral B toothpaste) against hidden resident mouth cavity microfora reveals Acidaminococcus sp.(11%,) Veillonella dispar (11%), Veillonella parvula (10%), Peptostreptococcus sp. (12%), Stomatococcus mucilaginous.(15%), Streptococcus ratti (11%), Veillonella atypical (8%), Streptococcus sangus (10%),  and Streptococcus mutans (12%), Percentage frequency distribution  of antibacterial activity of conventional toothpaste (MyMy toothpaste) against hidden resident mouth cavity microfora depicts Acidaminococcus sp.(12%), Veillonella dispar (9%), Veillonella parvula (8%), Peptostreptococcus sp.(10%), Stomatococcus mucilaginous.(16%), Streptococcus ratti (9%), Veillonella atypical (15%),Streptococcus sangus (9%) and Streptococcus mutans (12%), Percentage frequency distribution of antibacterial activity of conventional toothpaste (Olive toothpaste) against hidden resident mouth cavity microfora shows Acidaminococcus sp.(9%), Veillonella dispar (10%), Veillonella parvula (10%), Peptostreptococcus sp.(12%), Stomatococcus mucilaginous.(13%), Streptococcus ratti (10%) ,Veillonella atypical (17%), Streptococcus sangus (7%),  and Streptococcus mutans (12%). Pycnanthus Angolensis stem bark extract inhibited the growth of the oral bacterial isolates with of zones of inhibition diameter ranging from 6 mm to 17 mm at a concentration of 100mg/ml. Secondary metabolite (Phytochemical) screening shows the presence of flavonoids, tannins, saponins, alkaloids, reducing sugars, steroid, phenol, terpenoid, pyrrolozidine alkaloid, glycoside and cardiac glycoside with glycoside and terpenoid most present. However, anthraquinones and volatile oil were absent. With menial antibacterial activity, P. angolensis can be use in the formulation of herbal toothpaste. It should be advocated that Pycnanthus angolensis should be added to our convention toothpaste to improve the functional ingredient of the toothpaste and Plant-based traditional knowledge has become a recognized tool in search for new sources of drugs. It is clear that the use of these herbal plants can offer a platform for further research.

Open Access Original Research Article

Phenotypic and Genotypic Identification and Antifungal Susceptibility of Some Fungi Isolated from Tympanotonus fuscatus var. radula

Asemota Uwem Kelly, Obiekezie Smart Obuneme, Makut Makwin Danladi, Jubril Egwu Owuna

Asian Journal of Biochemistry, Genetics and Molecular Biology, Page 37-45
DOI: 10.9734/ajbgmb/2020/v6i230148

Aim: This study aimed to identify fungi isolated from Tympanotonus fuscatus var. radula and evaluate its level of susceptibility to known antifungal compounds.

Place and Duration of Study: Biotechnology Advanced Research Centre, Sheda Science and Technology Complex, Abuja between September and December 2019.

Methods: Tympanotonus fuscatus var. radula samples were purchased from the Keffi, Masaka, and Orange markets in Nasarawa State, Nigeria. Fungal isolation was achieved using Sabouraud dextrose agar supplemented with chloramphenicol and incubated at 28ᵒC for 5 days. ITS-1 and ITS-4 primers were used at 94°C for 2 min, 52°C for 1 min, and 72°C for 2 min for the polymerase chain reaction before sequencing at Inqaba Biotech South Africa. Disk diffusion technique was employed for antifungal susceptibility testing.

Results: Data obtained revealed that the suspected fungal species exhibited a generally higher level of resistance (19-40 mm) to 1 µg voriconazole in addition to a 20-35.5 mm zone of inhibition against 10 µg ketoconazole. Blast sequence analysis of the isolated samples revealed a 99.65% sequence homology to Meyerozyma guilliermondii, 99.38% to Fusarium oxysporium isolate E-225 1 and 96.23% to Aspergillus terreus isolate A2S4.  

Conclusion: Food safety involves isolating and accurately identifying disease causing pathogens such as fungi in food. Based on the fungal load obtained from this study, proper cooking and handling of sea-food which would otherwise cause disease, is highly recommended.

Open Access Original Research Article

Purification and Characterization of a Thermostable Chitinase Produced by a Fungus Isolated from Fruit Tree Rhizosphere

I. A. Afolayan, J. F. Oyun, E. A. Ekundayo, F. O. Ekundayo

Asian Journal of Biochemistry, Genetics and Molecular Biology, Page 46-56
DOI: 10.9734/ajbgmb/2020/v6i230149

This study reveals the chitinase producing ability of some fungi isolates cultured from the rhizosphere of different fruit trees such as mango, cassava, guava, and banana inside FUTA farm. A total of 16 isolates identified as Aspergillus nidulans, A. niger, A.flavus, A. fumigatus, A. ripens, Trichoderma viride, Mucor mucedo, Penicillium frequentans, Rhizopus stolonifer, Paecilomyces fumosoroseus, Gibbelula suffulfa and Geotrichum albidum were obtained and screened for chitinolytic activity. The effect of cultural conditions such as pH, temperature, metal ion, nitrogen source and carbon source was determined on Aspergillus nidulans, being the best chitinase producer. Further, the chitinase produced by Aspergillus nidulans was concentrated by ammonium sulphate precipitation and purified consecutively by gel filtration and ion-exchange chromatography. The optimum pH and temperature for chitinase activity and stability were examined as well as the effect of metal ions on the enzyme activity. The enzyme was most active at pH 7.0 and it was relatively stable at pH 4.0 - 9.0 retaining over 60% of initial activity after 120 min of incubation. The enzyme was most active at 50°C, possessing high thermal stability at high temperature of 70°C. The purified chitinase was significantly inactivated at 80°C and almost completely at 90°C when it was pre-incubated at these temperatures for 60 min. The enzyme was strongly inhibited by FeSO4, ZnCl2 and MnCl2 and was less sensitive to CaCl2 and KCl. This purified Aspergillus nidulans chitinase can be used as a catalyst for the degradation of chitin-containing compounds.

Open Access Original Research Article

Effect of Aqueous Extract of Costus afer Stems on the Serum Proteins and Bilirubin Levels of High Fat Diet Induced Hyperlipidemic Rats

Anyiam C. Chioma, Anyiam C. Albert, Onyeneho R. Ijeoma

Asian Journal of Biochemistry, Genetics and Molecular Biology, Page 57-64
DOI: 10.9734/ajbgmb/2020/v6i230150

Aim: The effect of aqueous extract of Costus afer stems on total protein, albumin,globulin, total and

conjugated bilirubin levels in diet induced hyperlipidemic rats were studied.

Methodology: Wistar albino male rats (100-135 g) were randomly distributed into 7 groups of 12 rats each. Group I was fed with standard diet as normal control rats and all the other groups were fed with high fat diet (10 g eggyolk/day) for 2 weeks. The plant extract was administered orally at different concentrations of 400, 800 and 1600mg/kg b.w alone and also in combination with the reference drug, Atorvastatin® (0.26mg) to the treatment groups for four weeks.  The serum proteins and bilirubin were observed at specific intervals (2 weeks).

Results: The results revealed significant (p<0.05) increase in the total protein, albumin and globulin concentration after 2 weeks of feeding with high fat diet in rats which were in groups IV and V as compared to normal control rats. Treatment of rats with plant extract showed no significant difference in the total protein concentration of hyperlipidemic test rats while the albumin concentration of rats in group VII increased significantly when compared to the normal and hyperlipidemic control rats. There was no significant difference in the globulin concentration of hyperlipidemic treated rats. After 2nd and 4th week of treatment, the total bilirubin concentrations of rats in groups V and VI (HTR on aqueous extract, 1600 mg/kg and HTR on aqueous extract, 800 mg/kg) decreased significantly when compared with normal and hyperlipidemic control rats. After 2 weeks of treatment, the conjugate bilirubin concentration of rats in group VI (HTR on aqueous extract, 1600 mg/kg) significantly (p<0.05) decreased when compared to the hyperlipidemic control rats.

Conclusions: Hence, this shows that Costus afer stem extract does not have any deleterious effect on tissues and on the analyzed parameters.