Open Access Original Research Article

Alkaline Cellulase Production by Penicillium mallochii LMB-HP37 Isolated from Soils of a Peruvian Rainforest

Karin Vega, Victor Sarmiento, Yvette Ludeña, Nadia Vera, Carmen Tamariz-Angeles, Gretty K. Villena, Marcel Gutiérrez-Correa

Biotechnology Journal International, Page 160-168
DOI: 10.9734/BBJ/2015/17831

Alkaline cellulases are demanded by the textile industry for several purposes but commercial preparations showing activity at alkaline conditions are very scarce.

Aim: To characterize a Penicillium strain isolated form soils of a Peruvian rainforest showing alkaline cellulase activity that may be useful for the textile industry.

Methodology: The molecular identification was based on the DNA sequence of its ITS region using ITS1 and ITS4 primers after PCR amplification. Cellulase production was evaluated in shaken flasks by using either lactose or microcrystalline cellulose. Total cellulase (as FPA) and endoglucanase activities were evaluated by the standard methods at several pH levels. Also, the cellulase activity of culture filtrates was tested for antipilling activity as compared to a commercial neutral cellulase preparation.

Results: After raw data of ITS DNA sequence was processed, multiple alignment and phylogenetic analysis confirmed that our strain can be named as Penicillium mallochii LMB-HP37. Higher activity was attained for neutral total cellulase on lactose (3371±108 U/l at pH 7.4) and alkaline cellulases attained similar activity levels than the acid cellulase (2978±151 U/l at pH 8.4 and 2910±42 U/l at pH 9.4). FPA and endoglucanase activities were produced at high volumetric (46.8±1.5 and 13.5±1.0 U/l.h, respectively) and specific (32.9±1.1 and 9.5±0.7 U/gbiomass.h, respectively) productivities at the same pHs which indicate that this strain may be suitable for commercial development. The enzyme of P. mallochii LMB-HP37 had slightly better results than the commercial enzyme as an anti-pilling agent even though is a crude preparation.

Conclusion: Penicillium mallochii LMB-HP37 produced high total cellulase activity on lactose which compares to well-known cellulase producers but at neutral to alkaline pH levels. Data obtained reveal that the crude enzyme is suitable for anti-pilling process (biopolishing) and may be also useful for biostoning.

Open Access Original Research Article

Highlighting the Degradation of Actin in Longissimus dorsi Muscle of Different Species: Bovine, Ovine, Caprine, Poultry and Freshwater Fish

Hasina Rasolofoharitseheno, Wisdom Michael Mompi, Yasmine Boudida, Mouhammed Gagaoua

Biotechnology Journal International, Page 169-176
DOI: 10.9734/BBJ/2015/17054

This research is aimed at highlighting the degradation of actin in the Longissimus dorsi muscle of five species, namely, bovine, ovine, caprine, poultry, and freshwater fish. To reveal the degradation of actin, by Western blotting, the myofibrillar proteins of the five studied species were first separated by using sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and colored with coomassie brilliant blue G-250. The colored polyacrylamide gel showed a similarity of the bands’ electrophoretic profile in bovine, ovine, and caprine. However, the bands observed in poultry and freshwater fish are specific for each species. The Western blotting revealed the presence of actin fragment of 32 kDa in bovine, ovine, and freshwater fish. A fragment of 28 kDa was found in the five studied species. However, a fragment of 19 kDa was obtained only in caprine. Two fragments of light molecular weights, 15 and 12 kDa, were found in four species such as bovine, ovine, caprine, and poultry. The fragments of 12 and 28 kDa were revealed for the first time by this research, where no previous study has announced their existence.

Open Access Original Research Article

Establishment of a Simple Plant Regeneration System Using Callus from Apomictic and Sexual Seeds of Guinea Grass (Panicum maximum)

Chen Lanzhuang, Nishimura Yoshiko, Umeki Kazuma, Zhang Jun, Xu Chengti

Biotechnology Journal International, Page 183-190
DOI: 10.9734/BBJ/2015/17215

Aims: For analysis of apomixis genes, as the first step, an efficient and simple plant regeneration system has been established using callus from apomictic and sexual seeds of guinea grass (Panicum maximum).

Study Design: The best basic medium for callus formation from matured seeds of guinea grass was selected, and then, the best combinations of growth regulators on different media were selected for plant regeneration by indirect organogenesis.

Methodology: Guinea grass accessions of sexual and apomict of seeds were used for culture. Seeds sterilized were cultured in Murashige and Skoog [10] medium (MS) and in that proposed by Chu et al. [12] (N6D) for callus formation. The best medium and, the effects of L-proline and growth regulators’ type and concentrations on callus formation and plant regeneration in 3 accessions were examined, respectively. After the plantlets rooting in MS hormone-free medium, the complete plants were planted in pots for hardening.

Results: N6D medium has given better rates for callus formation, that is, 97.1% in sexual N68/96-8-o-5, and 91.7% and 84.6% in apomict N68/96-8-o-11 and ‘Natsukaze’, respectively. MS medium with 0.2 mg/l of Kinetin has given the best rate or plant regeneration among the used 4 kinds of the media. For each material, the best results were obtained on MS with 0.2 mg/l of Kinetin for N68/96-8-o-5, and MS with 2.0 mg/l of L-proline and 0.2 mg/l of Kinetin for ‘Natsukaze’. After hardening of the regenerated plants in soil, 100% of surviving rates were obtained, and showed normal growth comparing with the plants-derived from seeds.

Conclusion: We have established, as the first case, an efficient and simple plant regeneration system by using callus from apomictic and sexual seeds of guinea grass for analysis of apomixis genes, consisting of analysis of the best media, L-proline usages and phytohormone combinations for callus formation, plant regeneration and hardening in different stages, respectively.

Open Access Original Research Article

Human Oral Lower Palate Isolates and their Antagonistic Effect on Phytopathogenic Fungi Pathogens

C. E. Aruwa, O. J. Adegoke, F. C. Adetuyi

Biotechnology Journal International, Page 191-199
DOI: 10.9734/BBJ/2015/17649

Production of certain substances that inhibit other microorganisms in the microbial environment of the oral cavity could serve as aggressive by product that may eliminate competitors and pathogens. Hence, this study was carried out to isolate and identify microorganisms from the oral cavity (lower palate of the mouth) and challenge these organisms with some selected strains of pathogenic fungi. The samples were aseptically collected using sterile swab sticks and transported to the Microbiology laboratory, for morphological, biochemical, and antagonistic tests in vitro. Nutrient Agar (NA) and MacConkey Agar (MCA) were used for the isolation of bacteria, Potato Dextrose Agar (PDA) for fungi and Malt Extract Agar (MEA) for antagonistic test. The isolated oral bacteria were Bacillus sp., Lactobacillus sp., Staphylococcus aureus and Streptococcus salivarius. Antagonistic effect was measured by zone of inhibition between the fungal plug and bacterial streak. The inhibition varied with different fungi. Results revealed that there were considerable variations in inhibitory activity. The zone of inhibition was more apparent in Bacillus sp. against Fusarium oxysporum (57.2±0.1:     P = .05). There was no inhibition/antagonistic activity with S. aureus and S. salivarius against all the selected fungi (0.0 - 0.1±0.1: P = .05). Antibiotics susceptibility test was carried out on the isolated oral microorganisms. The highest zone of inhibition was found in cotrimoxazole against Lactobacillus (23 mm), while the lowest zone of inhibition was found in ciprofloxacin against Lactobacillus sp. S. salivarius exhibited resistance to all antibiotics (0-12 mm). Bacillus sp., Lactobacillus sp. and S. aureus showed susceptibility to gentamycin. None of the bacterial isolates showed susceptibility to perfloxacin and streptomycin. Hence, gentamycin could be used to treat oral/dental infections caused by these bacterial isolates. Result from preliminary antagonistic studies showed that Bacillus sp. and Lactobacillus sp. could prove to be potent against all selected pathogenic fungi. Therefore, the metabolites produced by these isolates could be further studied for use as biocontrol agents of diseases caused by these fungi.

Open Access Review Article

Towards Efficient In vitro Regeneration of Cowpea (Vigna unguiculata L. Walp): A Review

Lawan Abdu Sani, Inuwa Shehu Usman, Muhammad Ishiaku Faguji, Sunusi Muhammad Bugaje

Biotechnology Journal International, Page 174-182
DOI: 10.9734/BBJ/2015/16841

Cowpea is a crop of tremendous economic and ecological values particularly in sub-Saharan Africa, where over 80% of the crop is produced and consumed. Due to heavy attack by pests and diseases, actual yield does not exceed 20% of the crop’s potential in most of the production regions. Shortages of genes to combat biotic stresses in the germplasm and sexual incompatibility with wild relatives are major impediments in cowpea improvement. Genetic modification of cowpea with relevant genes can address these problems. Establishment of reproducible regeneration system is a prerequisite for genetic transformation of cowpea using transgenic technology. Cowpea is among the most recalcitrant crops for manipulation under In vitro condition especially via de novo process. However, strategies to regenerate cowpea under In vitro conditions have evolved steadily in the last three decades. In this review, we give a summary of cowpea regeneration work carried out so far and discussed approaches employed as well as challenges of developing efficient regeneration systems in cowpea.