Research Journal of Biotechnology

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Model Predictive Control of Glucose Feeding for Fed-batch Candida utilis Biomass Production

Guo Qingqiang1,2, Liu Guoli1,3, Dong Nannan1,4, Li Qiqiang2, Lin Jianqun1 and Lin Jianqiang1*

In this work model predictive control (MPC) is used for control of glucose feeding rate of fed-batch culture for production of Candida utilis cell mass. A two layer hierarchical control system with the upper level for supervision and lower level for conventional control is designed which realizes automatic control, storage and refresh of kinetic model, data collection and on-line monitoring, data analysis and processing. The model stored in supervision computer is refreshed using genetic algorithm (GA) when the predictive error is over the limit. The results show that this method can control glucose concentration well and increase cell mass production effectively.

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Manganese ion transporter genes in Deinococcus radiodurans

Chang S.1,2 and Shu H.1,2,3*

Deinococcus radiodurans was one of the most radiation resistance microbes. There were two types of predicted Mn (II) transporters in D. radiodurans (Nramp family and ATP-dependent ABC-type transporter). DR1709 belonged to the Nramp family. DR2523 and DR2283-DR2284 were predicted ATP-dependent ABC-type transporters. Although some protein-tyrosine phosphatases can regulate the expression of Nramp gene, DR2161, one possible protein-tyrosine phosphatase, had no regulation role on the expression of DR1709. DR1708, a predicated secreted protein, had little role on the expression of DR1709 and the bacterium’s radiation resistance. DR1709 and DR2523 were parallel array. When there were sufficient nutrition in medium, DR2523 can play partial role of DR1709. But when manganese ions content was low, DR1709 may be the only gene responsible for manganese assimilating. The positions of DR2283 and DR2284 were tandem array.

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A novel bioprocessing debittering fermentation technology by Clavispora lusitaniae mutant

Sahota P.,* Sharma S. and Pandove G.

Naringinase is commercially attractive due to its potential usefulness in debittering citrus juices, so a novel bioprocessing debittering fermentation technology by naringinase producing strain Clavispora lusitaniae for the preparation of low-alcoholic naturally carbonated beverage from kinnow has been developed. Microbiological, physico¬chemical, mineral and sensory evaluation of kinnow beverage with 40 per cent juice revealed pH 3.4, TSS 12.6°B, acidity 0.54%, ascorbic acid 6.72 mg/100ml, reducing sugar 1.38%, total sugars 10.10%, limonin 3.3 ppm, naringin 161.9 ppm, total carotene 0.50 mg%, potassium 672.27 mg/kg, calcium 70.64 mg/kg, magnesium 61.89 mg/kg, sodium 19.24 mg/kg, iron 0.73 mg/kg, alcohol 0.86 (%v/v), CO2 1.36 bar and total plate count 3.3x108cfu/ml, ranked highest for taste (7.9), aroma (8.5), colour (7.8), astringency (8.25) and overall acceptability (7.8) during storage period of 3 months under refrigerated conditions (40C). The percentage decrease in limonin and naringin on storage was 54 and 64.8 percent, below the threshold level of limonin (6ppm) and naringin (600ppm) respectively. The major elements (K, Mg, Na, Ca, Fe) were retained in kinnow beverage during storage for period of 3 months. Carbon dioxide developed in the beverage acts as supercritical solvent, having antimicrobial property, imparts tangy taste, effervescence and has masking effect on bitterness.

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Role of β-Glucans and Levamisole on the Enhancement of the Survival Rate, Immune Response and Disease Resistance of Blue-Fin Porgy Sparidentax hasta (Sobaity) Larvae

Al-Gharabally H.*, Al-Marzouk A. and Azad I. S.

The immunostimulatory effects of two potential immunostimulants, β-Glucans (BG) and levamisole (LM) and their combination (MX) on the survival rate of blue-fin porgy Sparidentax hasta (sobaity) larvae were investigated. Also, different immune responses and disease resistance were evaluated in the present study. The optimum doses for β-Glucans and levamisole were determined to be 0.045 g/l and 0.3 g/l respectively. The β-Glucans, levamisole and their combination were evaluated in sobaity larvae via immersion and mixing with live feed for 30 d after hatching. The results showed that the immunostimulated larvae showed significant improvement in the growth rate compared to the non stimulated control larvae. Immune assays such as lysozyme activity, bacterial agglutinins and haemagglutinins were enhanced in sobaity larvae fed with immunostimulants. The use of immunostimulants, as a dietary supplement to sobaity larvae developed significant resistance to stress tests like salinity, transportation and handling. The protective response via the challenge test of the stimulated larvae was significantly (P<0.05) enhanced due to immunostimulation compared to the non stimulated larvae. These results suggest that the introduction of the two immunostimulants studied separately into the fish diet could increase their resistance to bacterial infection, reduce fish mortality rates and offer economic benefits.

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Identification of Human Protein Drug Targets Homologues with Data Mining

Feng Yanghe 1* and Wang Tengjiao 2

Identification and validation of potential target proteins is the first step for drug discovery and design. An accurate drug target classifier is helpful to test a new drug target more efficiently and economically. In this paper, we analyzed 522 drug targets and 5371 non-drug targets with 38 chemical and physical properties to identify differences on their chemical and physical properties. It shows the significant differences can be summarized into 9 properties. Based on these sequence features we used four data mining techniques to train drug target classifiers and gained lists of the potential target proteins. The results of 10 fold cross validation show that the accuracy of support vector machine (SVM) is 81.4% which is the highest in these classifiers. By integration of the lists predicted from our classifiers, a drug targets homologues set is identified to help drug discovery.

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Ecofriendly RO 13 Dye Decolorization by newly isolated Alcaligenes faecalis PMS-1 using Lignocellulosic Agriculture Waste

Shah P.D.

The aim of the present work was to develop ecofriendly decolorization process for cyanuric chloride based dye, Reactive Orange 13 (RO 13). Bacterial strain was isolated from dye-contaminated soil samples of local dye manufacturing industry. The bacterial isolate was identified as Alcaligenes faecalis PMS-1 with a NCBI accession number, GenBank ID: JF297973 based on 16S rDNA analysis. Decolorization experiments of RO 13 were carried out in Bushnell and Haas medium (BHM) in the presence of different fifteen carbon and seven organic nitrogen sources as well as optimum quantities were determined. RO 13 decolorization by PMS-1 was also studied in the presence of extract of agricultural by-products like rice husk, rice straw, sugarcane baggase powder and wood straw. In our study, successful replacement of conventional growth medium (nutrient broth) was demonstrated without compromise on rate and extent of decolorization along with less contribution of COD. Economically feasible and practically acceptable decolorization process for cyanuric chloride based reactive dyes was developed using lignocellulosic agriculture waste.

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Mycotoxin Production by Fusarium spp Isolates on Wheat Straw in Laboratory Condition

Matny Oadi Najim

This study was conducted to evaluate the capacity of 58 Fusarium spp isolates produce mycotoxin in infested wheat straw, the more popular feed for animal farm. Results showed that Fusarium spp grown in wheat straw, pre-incubated with Fusarium spp, isolated growth (dish of 1cm diam/10g) and incubated at 25 ±2 ºC for 21 days, produced many chemotyps. The highest toxin producers among Fusarium spp isolates were F.culmorum CS3716 which produced DON toxin at 9.07 mg/kg, F.culmorum CS3350 at 14 mg/kg T2-toxin, F.verticelloides IR 20 produced Fumonisin toxin at 10062.7 mg/kg and F. pseudograminearum CS3270 produced Zearalenone toxin at 211.9 mg/kg.

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Influence of silver nitrate on shoot regeneration from excised meristems of Momordica cymbalaria Hook.: a diminishing species

Balkhande S.V., Kure S.R. and Surwase B.S.*

The effect of silver nitrate (AgNO3) on shoot bud induction and proliferation from nodal explants derived from mature plants of M. cymbalaria Hook. was investigated. The nodal explants were cultured on Murashige and Skoog (MS) medium containing BAP, Kinetin alone or in combination with NAA, IBA and AgNO3 for multiple shoot induction. AgNO3 highly influenced the shoot bud formation and their subsequent proliferation. The best medium composition for multiple shoot induction was BAP (2 mgl-1) and AgNO3 (3 mgl-1) in MS medium. An average of 14.30±0.64 shoots were obtained per explant after 4 wk of culture. The number of shoots per explant increased three fold as compared to those formed on control. The rooting of in vitro developed shoots was best in MS ¼ medium with IBA (0.5 mgl-1). Micropropagated plants were successfully acclimatized (70 %) within 8 weeks after rooting. The standardized protocol reported in this study may help in the conservation of this species which is currently exploited from the nature.

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Pseudomonas stutzeri RP1: A Versatile Plant Growth promoting Endorhizospheric Bacteria inhabiting Sunflower (Helianthus annus)

Pandey R.,* Chavan P.N., Walokar N.M., Sharma N., Tripathi V. and Khetmalas M.B.

The extensive use of chemical fertilizer brought impressive gains in food production but with insufficient concern for sustainability. Crop plants are able to use about 50% of the applied fertilizer N while 25% is lost from the soil –plant system through leaching, volatilization, denitrification and due to many other factors causing not only an annual economic loss of US $ 3billion but also causes pollution to the environment.52 Plant growth promoting rhizobacteria (PGPR) can be defined as the indispensable part of rhizosphere biota that when grown in association with the host plants can stimulate the growth of the host. A Gram-negative, rod shaped bacterial isolate Pseudomonas stutzeri RP1was isolated from Sunflower endorhizosphere and was identified and characterized for plant growth promoting activities. PGPRs can be classified into extracellular plant growth promoting rhizobacteria (ePGPR) and intracellular plant growth promoting rhizobacteria (iPGPR)38. Plant growth promoting traits were analyzed by determining growth in nitrogen free medium, indole acetic acid (IAA), NH3 and HCN production and P- solubilization efficiency. The taxonomic position of the bacterium was confirmed by sequencing of 16S rRNA and phylogenetic tree analysis. The 16S rRNA gene sequence analysis confirmed the isolate to be Pseudomonas stutzeri. Further phylogenetic tree constructed with Clustal W2 showed its alignment with 16SrDNA sequence of Pseudomonads stutzeri ATCC17588 and P.stutzeri IARI-L-119. The strain showed antibacterial activity against E. coli, Xanthomonas sp, Serratia marscense, Bacillus cereus and Pseudomonas sp Isolate RP1 showed multiple plant growth-promoting attributes such as phosphate solubilization, IAA, NH3, HCN production and antibacterial activity. The present work demonstrates that the strain could be used as bioinoculant to attain the desired plant growth-promoting activity in plant growth promotion.

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Efficient Production of androstadienedione and testolactone from progesterone by biotransformation using Fusarium solani

Bing He1,2 and Wei Li1*

This study describes the efficient production of androstadienedione (androsta-1,4-diene-3,17-dione, ADD) and testolactone (1-dehydrotestololactone) from progesterone by biotransformation using the filamentous fungus Fusarium solani 14-2. These two metabolites produced in high yield were purified and were characterized on the basis of spectroscopic features (1HNMR, 13C NMR, IR and TOF-MS). The effects of some process conditions, like the pH value, concentration of substrate, solubilizing agents have been investigated. When tween-80 was used to solubilized progesterone with concentration of 10g/L, the substrate was completely consumed at 120 h and the yield of ADD reached its maximum at 120 h with concentration of 8.355 g/L and testolactone reached its maximum at 240 h with concentration of 8.682 g/L. The conversion rate of progesterone to androstadienedione and testolactone was not affected by progesterone concentration below 10 g/L. The optimum pH value for the bioconversion was 5.8. The results obtained in this study indicate that the strain of Fusarium solani 14-2 has the ability to convert a high concentration of progesterone to produce androstadienedione and testolactone in high yield and indicating its potential for commercial production of testolactone or ADD.

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Chemical and Laccase catalysed oxidation of gallic acid: etermination of kinetic parameters

Zinnai A., Venturi F.,* Sanmartin C., Quartacci M.F. and Andrich G.

During grape crushing, the release in the liquid phase of oxidizing enzymes can reduce the concentration of phenols which are natural antioxidants important for human health. On the contrary, in the following phases, related to the aging of wine, it is the slower chemical oxidation that causes the unwished degradation of the phenolic compounds. In spite of the wide literature on the field, kinetic evaluations which allow to quantify the extent of the phenomenon with temperature (30 and 45°C) are not yet available. To evaluate the phenol degradation kinetics, a model solution containing gallic acid which was subjected to both chemical and enzymatic oxidation was used. After determination of the kinetic equation able to describe the time reduction of the gallic acid concentration in the solution, the kinetic parameters related to the degradation curves and the oxidative process involved were calculated. In both the chemical and enzymatic oxidation experiments, an increase in the temperature of 15°C induced a reduction of the gallic acid oxidation rate, evidencing an influence of temperature on the oxygen amount dissolved in the liquid phase. In the presence of laccase, the oxidative kinetics was three orders of magnitude higher than that recorded without the enzyme.

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Antimicrobial compound from a novel Streptomyces termitum strain ATC-2 against Xanthomonas oryzae pv. oryzae

Jiang Donghua,1* Liu Qinying,1 Song Yiming 1 and Ji Hao 2

Bacterial blight caused by Xanthomonas oryzae pv. oryzae has been reported as one of the most destructive diseases of rice. An antagonistic strain ATC-2 against Xanthomonas oryzae pv. oryzae was screened and identified as Streptomyces termitum by biochemical and physiological assays which was reinforced by comparison of 16S rDNA sequence on Genebank via online BLAST analysis. The antimicrobial compound was isolated from the ferments broth by column chromatography over silica gel and Sephadex LH-20 and it is determined to be aloesaponarin II by 1H NMR and EI-MS analysis. The results of antimicrobial assays revealed that aloesaponarin II possessed strong antimicrobial activity against Xanthomonas oryzae pv. oryzae with IC50 19.2 μg/ml.

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Study of opsins as a colour producing factor in guppy fish (Poeciliareticulata)

Baqal Samreena and Jadhav Ujwala*

Guppy (Poecilia reticulata) fishes are renowned for their diversity and offer adaptation changes in the visual systems for spectral tuning.Tuning of visual system is typically accomplished through change in opsins development which includes cones and rods of eye.As colour plays significant role in mate choice, differences in visual sensitivities could greatly influence and even drive speciation. This fresh water fish has significantly different spectral sensitivities for cones present in eye retina. To establish whether the spectral qualities of environmental light affected the complement of visual pigment occur during ontogenetic process.Comparisons were made between fishes reared in three lighting aquarium conditions and between the natural environments (wild caught).The changes in spectral sensitivity of the cones were related to the differing levels of opsin development during developmental stages of eye.Significant differences were observed between development of cone sets of fishes living within the same environment and in different environment showing the changes in colour pattern.

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Can protease be an alternative agent for scouring of raw cotton fabrics?

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

Attempt has been made to analyse the applicability of bacterial protease as an alternative agent of scouring of raw cotton fabrics in place of sodium hydroxide to remove the natural impurities present in the fibre. Protease scouring shows lower weight loss values (4.0%) compared to the alkali scouring (6.15%) though no significant differences were observed in the drop absorbency values. Also, the proteases retain higher activity levels even after prolonged treatments at different pH values and temperature conditions. Proteases exhibit potential to replace harsh conditions employed in the scouring of cotton fabrics at present.

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Oxidative enzymatic gelation of sugar beet pectin for emulsion stabilization

Zaidel Dayang Norulfairuz Abang1,2 and Meyer Anne S.1*

Pectin from sugar beet is derived from the sugar beet pulp residue which results when sugar beets are processed for sucrose extraction. The sugar beet pectin has poor gelationability by the classic divalent-cation molecular mechanism because of a relatively high acetylation degree and short polygalacturonate backbone chain length. However, due to the feruloyl-substitutions on the side chains, the sugar beet pectic polysaccharides can be cross-linked via enzyme catalyzed oxidation. The enzyme kinetics and functionality of such oxidativelycross-linked sugar beet pectin, in relation to stabilizing emulsions has recently been investigated in model food emulsions. This paper reviews the pectin chemistry, enzymatic oxidative gelation mechanisms, interaction mechanisms of the sugar beet pectin with the emulsion droplets and explores how the gelation affects the rheology and stability of emulsion systems. The applied biotechnology concept of enzymatic gelation provides an array of opportunities for upgrading of low-value pectins for new food and non-food uses.

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