Research Journal of Biotechnology

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Research Journal of Biotechnology





Editorial

Biotechnological applications of venom toxins

Dhananjaya B.L.

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Effects of lactate and sulfate concentration ratios on the anaerobic mineralization of pentachlorophenol (PCP) in a combined PCP-dechlorinating and phenol-degrading culture

Yang S.,1 Inoue Y.2 and Katayama A .1,2 *

Anaerobic pentachlorophenol (PCP) mineralization achieved by combining two different anaerobic microbial populations (PCP-dechlorinating and phenol-degrading cultures) was studied according to different concentrations and ratios of lactate, as the electron donor for the dechlorination and sulfate as the electron acceptor for the anaerobic degradation. Complete PCP mineralization was found to be accelerated by providing the condition with an excess of sulfate for the complete consumption of lactate or its fermentation products. The concentration ratios of lactate to sulfate had a larger influence than absolute concentration on the PCP mineralization process. First-order kinetic analysis revealed that the sulfate-sufficient conditions enhanced the phenol degradation step but had no significant effect on the dechlorination steps. This information is critical for the design of permeable reactive barrier systems that use our anaerobic/anaerobic combination technology to remove PCP from the environment.

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Comparative Study of Bioethanol Production from Deoiled and Oiled Rice Bran

Tiwari Shubhra, Jadhav S.K.,* Tiwari K.L. and Esmil

On account of limited global supply of oil, bioethanol has emerged as an alternative for petroleum based liquid fuels. Nowadays, its use in automobiles as an alternative fuel has attracted worldwide attention for its production on a large scale while maintaining the economic status of a country. In present state of energy crisis, efforts are being made to reduce the dependence upon nonrenewable energy sources, one of which is bio ethanol, produced by fermentation of agricultural wastes and byproducts. An efficient ethanol production requires four components: fermentable carbohydrates, an efficient yeast strain, a few nutrients and simple culture conditions. Present study deals with bio ethanol production from de oiled and oiled rice bran by Saccharomyces cerevisiae. Comparative study of bio ethanol production and effect of incubation was also studied. Results indicate that maximum 8.58% of bio ethanol was produced from de oiled rice bran and 9.12% of bio ethanol was produced from oiled rice bran. Oiled rice bran is more efficient for bio ethanol production due to high carbohydrate content and fifth day was optimum incubation for better production.

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Response Surface Methodology Optimization of Process Parameters for Silage Production by Effective Microorganisms using Central Composite Design

Norfahana Abd-Talib1,2,3, Siti Hamidah Mohd-Setapar1,2*, Faizah Musa Nor1 and Kamal Khamis Aidee3

In this work, the effect of Activated Effective Microorganism (EM-A) on the quality of rice straw silages production have been studied by using Ash, crude protein, (CP) and crude fibre, (CF) levels as indicators. Treatments occur in two different types of silo (plastic bag and glass jar), different chopping lengths (1, 2, 4, 5 and 6cm) and different fermentation days (2, 12, 26, 40 and 50 days) to get the optimum performance of silage. Results showed that treatment with EM-A slowdown the degradation of CP content and decreased ash and CF content effectively compared to control rice straw. There were significant effects of type of silo on ash and CF content (P<0.05) and slightly significant on CP (P<0.10). No significant effect of fermentation day (P>0.10) was observed on ash, CP or CF content. The optimization point reached highest desirability of 0.660 in glass jar silo, 5cm chopping length and 40 days fermentation with highest CP recovery at 2.01% and lowest ash and CF content at 4.25% and 15.59% respectively. Highest desirability will be obtained from wider range of parameters used in silage production optimization study.

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Preparation and Antibacterial Activity of Silver Coated Activated Carbon from Rice Husks

Altintig Esra*, Bilgin Seyhan and Arabaci Gulnur

Silver has been in use since time because of its antibacterial effects. In the present century, the use of antibacterial metals in industry as disinfectant, by loading into inorganic materials is increasing and developing. But due to the emergence of several antibiotics the use of these silver compounds has decreased remarkably. In this study, activated carbons were produced from rice husks by chemical activation with zinc chloride (ZnCl2) at 700◦C in N2 atmosphere. Rice husk activated carbon supporting silver (RHC/Ag) was prepared by activation and chemical reduction. The RHC/Ag composites were characterized by SEM, XRD, BET and FTIR. Scanning electron microscopy (SEM) showed that the Ag particles were distributed uniformly on the rice husk (RHC) matrix. The antibacterial effects of characterized composites were also determined against Escherichia coli. A good antibacterial performance of the RHC/Ag composite powders was discovered against E.coli.

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Extra Cellular Acid Pectinase production from Three Different Bacillus Species and Their Bioscouring Efficiency in Cotton Fabrics

Senthilraja K.,1 Sreelakshmi S.N.,1 Vasanthi N.S.1 and Saravanan D.2*

An acidic, extra cellular pectinase from three species of Bacillus was optimized under submerged fermentation and its application tested for textile processing as a bioscouring agent in grey cotton fabric. The effectiveness of bioscouring was evaluated by measuring weight loss, wettability, crease recovery, ruthenium red dye test and FTIR. The results confirmed that pectinase from all the three species of Bacillus can be used as bioscouring agent of cotton fabrics. However, bioscouring with pectinase of B.licheniformis could be as much effective as the conventional alkaline scouring. The present study observed the efficacy of acidic, extracellular pectinase from three species of Bacillus viz., Bacillus subtilis, B.licheniformis and B.amyloliqufaciens to bioscour grey cotton fabric at pH 5 and 50°C temperature at a concentration of 1000U with a treatment time of 60 min and to remove pectin without damaging the fabric property by environmentally favorable method.

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Evaluation of Antioxidant Properties and Phytochemical analysis in the stem and leaves of Ceriops tagal mangroves

Jadhav B.L.,1* Quraishi Firdaus Mukhtar1 and Pagare B.G.2

The antioxidant activity and antioxidant phyto¬chemicals in the stem and leaves of Ceriops tagal species of Goa coast were studied. Present results have confirmed scavenging free radicals potential of C. tagal. IC50 values for DPPH radical scavenging of stem and leaves were 19.98 ug/mL and 29.97ug/mL respectively. The reducing power of both the parts of plant increased dose dependently in which the leaves showed higher reducing power. The HPTLC analysis of stem and leaves indicated the presence of flavonoids, anthraglycosides, bitter principle, saponins, terpenoids and essential oils. The antioxidant potential of C. tagal may be related to the presence of quality and quantity of these secondary metabolites.

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Genetic diversity and relationship among 40 rice accessions from North-Eastern Zone of Tamil Nadu using morphological and SSR markers

Thenmozhi P.1,2 and Rajasekaran C.2*

The present investigation was focused on morphological and molecular characterization of 40 rice accessions collected from North-Eastern Zone of Tamil Nadu. Morphological characterization using 14 quantitative traits was done in a field experiment in a randomized block design. The aim of the work was to study the genetic diversity among closely related genotypes using Morphological and Simple Sequence Repeats (SSR) markers and to select accessions that can be used as potential parents in a future breeding program. The dendrograms based on morphological traits and SSR markers were used to group the accessions. Genetic relatedness among accessions based on SSR molecular marker data was presented in the form of a dendrogram using the unweighted Pair Group Method with Arithmetic mean (UPGMA). Relative effectiveness of the SSR markers and genetic diversity among accessions as revealed by botanical descriptors were compared. The UPGMA classified the 40 accessions into nine morphological clusters whereas the SSR markers classified the 40 accessions into 12 cluster groups. Under morphological clustering ADT 36 and Poongar, Kullakar were the most distantly related and under SSR markers ADT 36 and TKM 3 are more distantly related. Hence these distinct accessions should be used in future breeding programs for improving yield and quality characteristics of rice. SSR markers free from environmental influence, were more discriminatory and informative as they were able to differentiate more pairs of accessions than the botanical descriptors.

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In vitro protocol for rapid mass propagation of Solanum stramoenifolium, a medicinal plant

Bukkambudhi Krishnaswamy Manjunatha,* Shinduja R. and Rashmi K. P.

An efficient propagation and regeneration system via indirect and direct shoot organogenesis for a medicinal plant Solanum stramoenifolium was established. High activity cytokinins (BAP) could directly induce adventitious multiple shoots from nodal explants within 3 weeks of culture. Indirect organogenesis through callus was successfully achieved in the cultures supplemented with auxin followed by shoot induction and root induction. Nodal explants when cultured on 4 µM BAP induced maximum number of shoots (19.5±0.16.)/explants within 3 weeks of incubation, microshoots started rooting in the same medium on prolonged culture (4-5 weeks). In vitro regenerated plants were successfully acclimatized. The leaf and petiole explants cultured on MS medium supplemented with 2µM 2-4 D induced proliferative green callus within 4 weeks of incubation. Proliferating callus on subculturing to MS media supplemented with 2 µM BAP induced shoot induction, microshoots transferred to culture supplemented with 1.0 µM IAA induced roots. Explants cultured on media supplemented with both cytokinin and auxin combination failed to induce expected results. Over 95% of plantlets survived following acclimatization and transfer to a potting mixture (1:1, sand:vermiculite) in basins.

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Efficient degradation of raw chicken feather into soluble peptides and free amino acids by a newly isolated Bacillus subtilis S1-4

Yong Bin, Yang Bin-Qing and Feng H.*

Feather is a byproduct by the poultry industry with huge annual yield. In view of its application, keratinolytic microorganisms have been employed to degrade feather. In order to achieve this purpose, a bacterial strain (Bacillus subtilis S1-4) adhered to the native chicken feather was isolated and identified by comparative genome analysis. This strain in simple basal salt cultures could completely degrade raw chicken feathers at 5%. The contents of soluble peptides and amino acids released from the feathers by strain S1-4 could reach 11.61 mg/ml and 6.86 mg/ml respectively. The strain S1-4 needed a narrow pH to achieve efficient degradation. In general, extra carbon and nitrogen sources did not improve the feather degradation. B. subtilis S1-4 could secrete several proteases and keratinase as well as the disulfite reductase activity, indicating that all of these enzymes are involved in the feather degradation. In addition, the effects of several parameters on keratinase production were primarily investigated. However, the optimal cultivating parameters for the mass loss of substrate and keratinase production were different suggesting that optimization of these parameters for various purposes should be performed separately. Conclusively, these results demonstrated that B. subtilis S1-4 is most promising in application to feather degradation.

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Purification and Characterization of Laccase from newly isolated Thermophilic Brevibacillus sp. (Z1) and its applications in removal of Textile Dyes

Ceyda Bozoglu1, Ahmet Adiguzel1*, Hayrunnisa Nadaroglu2*, Derya Yanmis3 and Medine Gulluce3

Laccases are multicopper-containing oxidases (EC 1.10.3.2.) and they are widely found in fungi, higher plants and bacteria. Because of their high pH stability and thermostability, laccases from this strain have a potential for the treatment of textile bleaching effluents. In this research, the thermophilic test strain isolated from the Agri-Diyadin thermal springs, in the east of Turkey, were subjected to analysis of 16S rRNA sequence. The obtained sequence data were compared with data from the database in the Genebank and as a result it has been identified that this organism is a member of Brevibacillus genus (GenBank number KC292196). The laccase from Brevibacillus sp. (Z1) was purified by using precipitate of saturation (NH4)2SO4, DEAE-cellulose and Sephacryl S200. The purification degree and the molecular mass of the enzyme (93 and 110 kDa) were determined by SDS-PAGE and gel filtration chromatography. In order to determine the enzyme's resistance against the metal ions, the effect of metal ions such as Ca2+, Ba2+, Mn2+, Ni2+, Co2+, Cu2+, Zn2+, Fe2+, K+ and certain chemical substances such as EDTA and ascorbic acid on the pure laccase enzyme activity have been investigated and no significant change was observed. Vmax and Km values were calculated using Lineweaver Burk plots for ABTS, 2,6-dimetoxyphenol and guaiacol. Purified laccase enzyme was also used to remove some textile dyes which are used in textile industries and known to cause major environmental problems such as Reactive Black 5, Fuchsine, Allura Red and Acid Red 37 from waste water. Consequently, it is concluded that the purified laccase from Brevibacillus sp. (Z1) can successfully be used for the removal of the textile dyes from wastewater.

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Enhancement of laccase production and biotechnological application of laccase pleurotus florida

Jenitta Emima Packiyam E.1 and Ragunathan R.2*

Laccases are oxidoreductases belonging to the multinuclear copper-containing oxidases; they catalyze the monoelectronic oxidation of substrates at the expense of molecular oxygen. Laccases are multi-copper oxidation of a wide range of inorganic substances coupled with one four electron reduction of oxygen to water. In this work laccase was produced from Pleurotus florida by using two different production media (media I and II) and production media II was found to be the best among two. All the laccase enzymes are extracellular glycoproteins and many of these enzymes show considerable heterogeneity after purification, so the enzyme is purified and parameters such as optimum day substrate concentration are characterized. Carbon and nitrogen sources in the medium play an important role in ligninolytic enzyme production, so effective carbon and nitrogen sources for optimum production of laccase was also found out. The molecular mass of lacasse isozymes varies from 50 to 100KDa. The molecular mass of partially purified lacasse was determined by using SDS-PAGE technique. Number of industrial applications for laccases has been proposed. In this study biopulping of saw dust using isolated laccase was carried out and kappa number of bleached pulp was obtained.

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Molecular characterization of free living nitrogen fixing bacteria associated with aerobic rice root zone soil and their influence on growth and biomass of Anethum graveolens

Earanna N.,* Nandini K. and Karaba N. Nataraja

Four free living N2 fixing bacteria were isolated from root zone soil of aerobic (upland rice) rice and identified by using 16S rRNA gene sequence. The gene sequence showed 97-99 percent homology for Azotobacter species/strains. Based on gene sequence homology (BLAST) they are identified as A. tropicalis, A. vinelandii DJ, A. vinelandii ISSDS 386 and A. vinelandii ISSDS 436. Influence of these strains either singly or in combination with the arbuscular mycorrhiza (Glomus intraradices) on growth and herbage yield of an aromatic culinary herb Anethum graveolens was studied in pot culture under glass house conditions. Results indicated that all the inoculated treatments either single or in combination with the G. intraradices increased the growth, herbage yield and nitrogen content compared to control. Among the four Azotobacters, the Azotobacter vinelandii ISSDS 436 strain produced significantly highest biomass and nitrogen content which is followed by A.vinelandii ISSDS 386 indicating their efficiency of nitrogen supplement to the plant.

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Therapeutic Effects of Human Epidermal Growth Factor

Oldooz Atashparvar*, Kumar Neethu S. and Seyed Amirali Raeisosadat

The discovery of EGF protein has opened new fields of widespread importance to basic science. EGF protein is well known for use in the cosmetic field but is also of major benefit in the management of gastric ulcer healing, elimination of skin scars, hastening the healing process of epidermis damage on skin etc. Human EGF protein stimulates dimerization of epidermal growth factor receptor by attaching to the extracellular region of the EGF receptor and forms EGF-EGFR complex. After binding the EGF to the EGF receptor, dimerization of EGFR stimulates tyrosine kinase activity thus promoting various downstream signaling pathways by EGF receptor tyrosine kinase to regulate proliferation and differentiation. Nowadays, since the EGF protein activates the cells for multiply; it has been widely used for treatment of wound and cancer in gene therapy and cancer vaccine.

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Plant Response to Abiotic Stresses: From the Perspectives of Gene Expression

Haruna Sa’idu and Sulaiman Mohammed*

The response to abiotic stresses by some selected plants species from the perspective of gene expression was reviewed. The plants selected are of highly valued in terms of agriculture, the utility of the plants as a food product has also been extensively described. Several abiotic stress factors are considered, some can be recognized easily. However, others are difficult to identify especially those which affect the environment and scientists believe that these factors will increase in the future due to the harsh changes in climate, known as global warming. Genes are chosen to review their impact at transcriptional levels under abiotic stresses and resistance to various plants.

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