Research Journal of Biotechnology

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Research Journal of Biotechnology





Study on accumulation feature of rhynchophylline and isorhynchophylline in Uncaria rhynchophylla

Qianqian Guo, Xiaojun Ma, Jin'e Fu and Wei Zu

The rhynchophylline (RIN) and isorhynchophylline (IRN) of Uncaria rhynchophylla had the effect to relieve and cure several diseases such as the Alzheimer's disease. However, the low yield of these natural products restricted their application in medicine. Synthesis physiology research of the RIN and IRN had positive significance on the development of their medicinal potential. In this study, we have measured the RIN and IRN content of hook stem (as the medicinal part), stem, leaf and capsule in the uncaria using the high performance liquid chromatography. The results showed that the capsule had the highest RIN and IRN content followed by hook stem and stem and the leaf had lowest RIN and IRN content respectively. Uncaria branches were divided into vegetative and reproductive growth branches. There were different accumulation change trends of the RIN and IRN in different parts of the vegetative-growth branches and reproductive-growth branches. The change of RIN and IRN content in the reproductive-growth branches showed the uni-modal curve respectively while that in those in the vegetative-growth branches had no significant change. RIN and IRN content of the leaf R, stem R and capsule between the reproductive-growth branches and the corresponding parts of the vegetative-growth branches showed the negative correlation relationship respectively. In addition, the ratio change between RIN and IRN in the stem (hook stem, stem V and stem R) was similar to that in the capsule. RIN and IRN content between the capsule and the stem R showed the significant positive correlation relationship. Not only the understanding of the RIN and IRN accumulation law provided reference for the selection of the research material on the study of uncaria physiology and molecular biology but also it laid foundation on the research of the RIN and IRN synthesis mechanism.

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Bacterial diversity in the fermented yellow water from the traditional Chinese liquor fermentation process

Zhang Qing, Li Ke, Lin Kai and Xiang Wenliang

Culture-dependent and molecular identification methods were employed to investigate the bacterial diversity in fermented yellow water from the fermentation process of the intense fragrance liquor. The result indicated that the dominant bacterial species in the fermented yellow water were Bacillus humi (19%), Bacillus bataviensis (11%) and Lysinibacillus sphaericus (17%), while they did not show any notable correlations with the liquor fermentation. However, five species which were Bacillus subtilis (5%), Bacillus amyloliquefaciens (2%), Bacillus altitudinis (1%), Lysinibacillus xylanilyticus (4%) and Solibacillus silvestris (2%) might response for corn degradation and esterification of flavor components, implying their important functions in liquor making. The H diversity index and the simpson dominancy index respectively revealed 1.83 and 4.50 respectively.

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The Research of Breeding High Yield Streptomyces roseosporus Strains with Protoplast Fusion

Сhen Guosheng

We exposed the daptomycin producing strain Streptomyces roseosporus D02 for 8min under 30W UV lamp (wavelength: 253nm; irradiation distance: 30cm) to inactivate it and used heating at 750C for 60min to inactivate the protoplasts of Streptomyces roseosporus D04 and made the above two inactivated protoplasts to fuse using PEG4000 as the fusogen. Then we screened out the regeneratable fusants from which we obtained a high yield S. roseosporus strain D07. In shake-flask fermentation adding capric acid, the daptomycin production is 91.72 mg/L which is 2.03 times and 2.01 times the yield of the two original strains respectively. This study provides the foundation for further related studies of Streptomyces roseosporus with genome shuffling.

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Evaluation of silver nanoparticles-chitosan encapsulated synthetic herbicide paraquate (AgNp-CS-PQ) preparation for the controlled release and improved herbicidal activity against Eichhornia crassipes

Karthick Raja Namasivayam S., Aruna A. and Gokila

Nanotechnology is currently employed as a tool to explore the darkest avenues of various fields of science and technology due to their amenability to biological functionalization. Nanoformulation of various agrochemicals is now extensively utilized for controlled and improved delivery. In the present study, silver nanoparticles-chitosan encapsulated paraquate nanoformulation was evaluated against controlled release and improved herbicidal activity against Eichhornia crassipes. Nano formulated herbicide was prepared by chemical reduction of silver nitrate with sodium borohydride and the obtained silver nanoparticles were mixed with chitosan and the field concentration of paaquate. Synthesized nanoformulation was characterized by scanning electron microscopy (SEM) which reveals uniform spherical particles with the size of 100nm and all the elemental composition of nanoformulation was confirmed by energy dispersive X ray spectroscopy (EDX) and X ray diffraction (XRD). Encapsulation efficacy was found to be 89.0%. Controlled release study showed 90.0% of release at 24 hours and the distinct improved herbicidal activity was observed against Eichhornia crassipes by the formation of necrotic lesions in all the tested concentration of nanoformulation. Non target effect study on the various soil parameters revealed that nanoformulation did not affect the soil macro and micronutrients, soil enzyme and soil microflora. Phytotoxicity of the nanoformulation was also studied using Vigna mungo. Seedling emergence and plant growth parameters of the tested plant were not affected in the nanoformulation treatment. The present study suggests the possible utilization of nanoformulated herbicide to control the economic important weeds.

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Isolation and Identification of New Species of Thermophilic Cyanobacteria and Bioremediation investigating of Heavy Metals

Ebli Farzad mohammadi, Heshmatipour Zoheir, Khanafarin Anita, Saffari Javad Roodgar and Imani Saber

A large family of bacteria is Cyanobacteria which have a large variety of types and have been found in various habitats. Heavy metals such as copper, lead and zinc-as a large group of elements are considered as toxic factors present in such environment as wastewater and soil. Some bacteria, Cyanobacteria for example, have the ability to remove heavy metals. The purpose of this study is isolation and identification of new species of thermophilic Cyanobacteria and investigating its ability to remove heavy metals. First, the collected samples were inoculated into BG11 broth medium. Then the masses were transferred to the BG11 agar medium and were incubated for 8-10 days. Finally, molecular diagnosis 16s RNA was developed on the colonies. Thermophilic Cyanobacteria were inoculated in medium containing copper, lead and zinc; results were calculated by atomic adsorption. In this study, we succeeded in isolating unculturable of thermophilic Cyanobacteria EBLI1 (89.2%) that was morphologically similar to oscillatoria and lyngbia. Cyanobacteri have also been identified as capable of elimination of 2.5% zinc, 13.5% copper and 27% lead from the media. This paper studies the bioremediation of copper (Cu), plumb (Pb) and Zinc (Zn) at different concentrations and under different conditions by Bacteria. The study has proved that heavy metals such as zinc, copper and lead are not toxic for Cyanobacteria, but they can reduce GT time bacteria.

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Separation of Bacillus thuringiensis from fermentation broth using microfiltration: Optimization approach

Rasoul Marzban, Fatemeh Saberi and Mohammad Mahdi A. Shirazi

In this work, the separation of spore and crystal from fermentation broth of Bacillus thuringiensis (Bt) via cross-flow microfiltration (MF) was evaluated. A commercial microporous membrane made of cellulose acetate (CA) with 0.45 µm pore size was used for experiments. Taguchi method was used for experimental design. Under optimal conditions, the permeation flux could be greater than 51 kg/m2.h. Results indicated that the spore and crystal proteins recoveries were found to be more than 99%. The results demonstrated that MF was effective for Bt based biopesticide recovery from fermention broth.

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Expression of napA, nirK, cnorB and nosZ in Chelatococcus daeguensis TAD1 at high temperature under oxic conditions

Yang Yunlong and Huang Shaobin

Pure cultures of Chelatococcus daeguensis TAD1 were incubated at high temperature (500C) using nitrate as a substrate and an electron acceptor for denitrification. The periplasmic nitrate reductase gene (napA), nitrite reductase gene (nirK), nitric oxide reductase gene (cnorB) and nitrous oxide reductase gene (nosZ) expression were measured using a quantitative reverse transcription-PCR at various initial dissolved oxygen (DO) concentrations. All the four genes expression reached a maximum at 12h. The maximum expression level of napA, nirK and nosZ was at 50rpm, the corresponding DO concentrations of which were 2.2 mg/l, but the maximum expression level of cnorB was at 150rpm, the corresponding DO concentrations of which were 2.6 mg/l. These results demonstrated that low dissolved oxygen conditions were more beneficial to expression of the denitrification genes in C. daeguensis TAD1 at high temperature.

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Marker-Trait Association for Fertility Restoration in Hybrids of Rice (Oryza sativa L.) under Aerobic Conditions

Reddy Kamalnath K. R., Kaki Nagendra, Varma Mohan Kumar C., Patil Kalmeshwar Gouda, Payasi Devendra , Anantha M. S., Shashidhar H. E. and Shenoy Vinay

A study was conducted to validate earlier reported SSR markers closely associated with fertility restoration (Rf gene) of WA-CMS lines in rice. Thirteen SSR markers across three chromosomes (chr.1, 10 and 12) were evaluated among 21 drought tolerant restorer lines. These lines were crossed to five different CMS lines to obtain 105 F1s. Single Marker Analysis was performed to determine the association between marker and fertility restoration in hybrids. Out of thirteen markers, RM6100 marker linked to Rf4 gene on chromosome 10 has shown significant association under both aerobic and wetland conditions in two consecutive Kharif (2010 and 2011) seasons, followed by other markers viz. RM171 linked to Rf4 gene RM258 linked to Rf4 and Rf6 genes all on chromosome 10. These markers will help facilitate marker assisted selection for identification of restorer lines in CMS-WA system. Molecular diversity analysis for twenty one genotypes along with known restorers using thirteen markers was carried out and the dendrogram revealed four clusters. Cluster-II and Cluster-IV consisted of single genotypes and Cluster-I consisted of 7 genotypes and Cluster-III consisted of 14 genotypes. Combined ANOVA for spikelet fertility revealed significant variation for season, moisture regime and genotypes.

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Enhancement of Skin Whitening Activities of Aronia melanocarpa by Ultrasonication Extraction Process at Low Temperature

Jeong Hwan Kim, Nam Young Kim and Hyeon Yong Lee

The ultrasonication process (UE) was employed to efficiently extract relatively hard fruits of A. melanocarpa (Aronia) by comparing the results from conventional extractions: hot water (WE), 70% ethanol extractions (EE). 104.5 mg ml-1 of the highest total polyphenol content was observed from the UE process as well as high amounts of two anthocyanins in A. melanocarpa such as cyanidin and dephinidin which should result in the best antioxidant activities as 70.93 % in DPPH free radical scavenging ability. Approximately 70% inhibition of melanin synthesis was also observed in adding the extracts from the UE which was also highest inhibition. It was also found that the whitening activities of the extract from the UE seemed to be better than those of each of components like cyanidin and dephinidin which implies that the extract might be effective in maintaining its biological activities rather than using each of single components, possibly due to synergistic effects of the bioactive substances in the extracts. It was also first shown that the whitening activities of the extract from the UE process were possibly associated with strong antioxidant activities of A. melanocarpa.

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Phytochemical Testing, Antioxidant Activity, HPTLC and FTIR Analysis of Antidiabetic Plants Nigella sativa, Eugenia jambolana, Andrographis paniculata and Gymnema sylvestre

Sundaramoorthy Sangeetha, Rastogi Archit and Arunachalam SathiaVelu

Earlier, we have investigated and proven the in vitro anti-diabetic effects of Nigella sativa, Eugenia jambolana, Andrographis paniculata and Gymnema sylvestre. In this paper, we discuss the results of their phytochemical analysis, antioxidant activity tests and HPTLC investigation. On phytochemical analysis, we found that all the plants, apart from containing the expected carbohydrates and proteins, also contained terpenoids, flavonoids and tannins. Though Andrographis paniculata tested negative for tannins, it was found to contain phlobatannins, a special class of tannins. HPTLC confirmed these results. We found Nigella sativa contained the highest number of flavonoids and all the four plants contained phenolic compounds. On literature review, we found that flavonoids are responsible for antioxidant activity and tannins for antidiabetic activity. Our antioxidant analysis results echoed this. Nigella sativa had a high antioxidant activity attributed to its excessive flavonoids. On FTIR we found all plants contained aromatic compounds and phenolic compounds. Most of the compounds were identical in all the four plants. A notable exception was alkenes found in Nigella sativa at wavenumber 989.48 cm-1 which might play a role in the antioxidant activity of the plant.

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The Effect of Aspect Factor in Isparta-Arapdağı Rangeland on the Dry Forage Yield and Botanical Composition

Babalık Ahmet Alper

The objective of this study is to determine the plant-covered area, dry forage yield and botanical composition in Isparta-Arapdağı rangeland during 2008-2009. The measurements were carried out for a period of two years during the months of June and September at 4 different aspects of the grazed and protected rangeland areas. “Line intercept (transect)” method was used for the determination of plant-covered area whereas “quadrat method” was used to determine dry forage yield. It was determined as a result of measurements that the average plant-covered area of the rangeland was 17.8 % whereas the average dry forage yield was 78.8 kg/da. The highest dry forage yield was determined at the northern aspect and the lowest was determined at the southern aspect. It was determined that Poaceae family comprises 48.0 %, Fabaceae family comprises 8.8 % and plants from the remaining families comprise 43.2 % of the botanical composition. Statistically significant differences were determined between the usage status, seasons, aspects and years when measurements were carried out in terms of dry forage yield.

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Utilization of poultry waste (chicken manure) for Cost effective and high density culture of two freshwater cyclopoid copepods Thermocyclops decipiens and Mesocyclops aspericornis

Ali Hyder, Mehrajuddin War, Naveed Saquib and Altaff Kareem

The benefits of using copepods in aquaculture include their superior nutritional value, high digestibility, movement patterns which trigger strong feeding response, small size and simply the fact that they are part of natural food chain. Two cyclopoid copepod, Thermocyclops decipiens and Mesocyclops aspericornis were mass cultured using chicken manure as feed for a period of 21 days. Adult T. decipiens occurred in high density on 14th day while high densities of copepodid and nauplii were recorded on 21st and 7th day of the culture period respectively. Higher densities of Mesocyclops aspericornis adult, copepodid and nauplii were obtained on 14th day (1845.55 ± 43.04), 21st (2458.88 ± 17.88) and 7th (1077.77 ± 4.84) respectively. The water analysis showed that the temperature range of 27 - 31ºC and pH of 6 - 7 was conducive for optimal growth of these two copepod species. The main focus of this approach was to check feasibility of using chicken manure for fertilizing the medium to grow high population densities of these species cost effectively. Our results show that by using chicken manure it is possible to culture these two species to high densities with proper feeding dosages.

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Thermostability of the xylanase from Streptomyces rameus L2001 and its use in xylooligosaccharide production

Li Xiuting and Zhu Yunping

Xylooligosaccharide was a promising ingredient in functional foods due to its various beneficial health effects. The thermostability of the xylanase from Streptomyces rameus L2001 and its use in xylooligosacchride using corncob were investigated in current study. The half-time (t1/2) of xylanase was determined at different temperatures. The thermoinactivation of the enzyme followed first-order kinetics and the energy of denaturation (Ea) was 230.082 KJ/mol. In order to improve the yield of xylooligosacchride, several factors affecting the hydrolysis reaction were examined including the enzyme dosage, substrate concentration and five surfactants (Tween 60, Tween 80, Triton 100, oleic acid and SDS). The optimal enzyme dosage and substrate concentration was 20U/g and 3% respectively. Non-ionic surfactants could effectively improve the hydrolysis yield with the addition of 5 g/L of Tween 80. The main components of the hydrolysis prodcuts were xylobiose and xylotriose by HPLC analysis which indicated that hydrolysis of the corncob xylan using the xyanlase from Streptomyces rameus L2001 was a promising method to produce xylooligosacchride.

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