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Preparative Biochemistry & Biotechnology

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https://www.readbyqxmd.com/read/28296566/a-new-elisa-based-method-for-detection-of-o-glcnac-transferase-activity-in-vitro
#1
Jieqiong Qi, Ruihong Wang, Yazhen Zeng, Wengong Yu, Yuchao Gu
O-GlcNAcylation is a dynamic, reversible, post-translational modification that regulates many cellular processes. O-GlcNAc transferase (OGT) is the sole enzyme transferring N-acetylglucosamine from UDP-GlcNAc to selected Ser/Thr residues of cytoplasm and nucleus proteins. Aberrant of OGT activity is associated with a number of diseases, suggesting OGT as a novel therapeutic target. In this study, we created a new ELISA-based method for detection of OGT activity. First, CKII, a well-known OGT substrate, was coated onto ELISA plate...
March 15, 2017: Preparative Biochemistry & Biotechnology
https://www.readbyqxmd.com/read/28281888/medium-ph-in-submerged-cultivation-modulates-differences-in-the-intracellular-protein-profile-of-fusarium-oxysporum-urm-7401
#2
Nathália Gonsales da Rosa-Garzon, Hélen Julie Laure, Cristina Maria de Souza-Motta, José César Rosa, Hamilton Cabral
Fusarium oxysporum is a filamentous fungus that damages a wide range of plants and thus causes severe crop losses. In fungal pathogens, the genes and proteins involved in virulence are known to be controlled by environmental pH. Here, we report the influence of culture-medium pH (5, 6, 7, and 8) on the production of degradative enzymes involved in the pathogenesis of F. oxysporum URM 7401 and on the 2D-electrophoresis profile of intracellular proteins in this fungus. F. oxysporum URM 7401 was grown in acidic, neutral, and alkaline culture media in a submerged bioprocess...
March 10, 2017: Preparative Biochemistry & Biotechnology
https://www.readbyqxmd.com/read/28281882/efficient-expression-of-stable-recombinant-human-insulin-like-growth-factor-1-fusion-with-human-serum-albumin-in-chinese-hamster-ovary-cells
#3
Aini Wan, Dongsheng Xu, Kedong Liu, Lin Peng, Yanfei Cai, Yun Chen, Yang He, Jianfeng Yang, Jian Jin, Huazhong Li
Insulin-like growth factor-1 (IGF-1) plays a crucial role in cell development, differentiation and metabolism and has been a potential therapeutic agent for many diseases. Chinese hamster ovary (CHO) cells are widely used for production of recombinant therapeutic proteins, but the expression level of IGF-1 in CHO cells is very low (1500 µg/L) and the half-life of IGF-1 in blood circulation is only 4.5 min according to previously studies. Therefore, IGF-1 was fused to long-circulating serum protein HSA and expressed in CHO cells...
March 10, 2017: Preparative Biochemistry & Biotechnology
https://www.readbyqxmd.com/read/28277946/separation-of-bsa-through-fau-type-zeolite-ceramic-composite-membrane-formed-on-tubular-ceramic-support-optimization-of-process-parameters-by-hybrid-response-surface-methodology-and-bi-objective-genetic-algorithm
#4
R Vinoth Kumar, I Ganesh Moorthy, G Pugazhenthi
In this study, Faujasite (FAU) zeolite was coated on low cost tubular ceramic support as a separating layer via hydrothermal route. The mixture of silicate and aluminate solutions was used to create a zeolitic separation layer on the support. The prepared zeolite ceramic-composite membrane was characterized by using X-ray powder diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), particle size distribution (PSD), Field emission scanning electron microscopy (FESEM) and zeta potential measurements...
March 9, 2017: Preparative Biochemistry & Biotechnology
https://www.readbyqxmd.com/read/28277945/polymer-membrane-ion-selective-electrodes-as-a-convenient-tool-for-lipases-and-esterases-assays
#5
Maciej Cieplak, Ryszard Ostaszewski
We have proposed a novel assay for lipases and esterases activity determination based on potentiometry with ion-selective electrodes (ISEs). Enzyme preparations, obtained from the living cells, are complex mixtures of various proteins, short peptides, lipids, carbohydrates and other compounds. The most commonly used quantitative methods in enzyme studies are based on spectrophotometric or spectroflourimetric protocols which has significant limitations. They are not valid for samples that are turbid or strongly colored...
March 9, 2017: Preparative Biochemistry & Biotechnology
https://www.readbyqxmd.com/read/28277822/comparison-of-dna-isolation-using-salting-out-procedure-and-automated-isolation-magna-system
#6
Marta Kalousová, Kateřina Levová, Aleš A Kuběna, Marie Jáchymová, Věra Franková, Tomáš Zima
INTRODUCTION: Isolation of genomic DNA is a key step in genetic analysis. The aim of the study was to evaluate the suitability of isolation of DNA from peripheral blood with manual salting out procedure and automated MagNA system under specific conditions. EXPERIMENTAL: The impact of storage conditions, type of material (whole blood or blood cells) and method used for DNA extraction were evaluated in terms of DNA yield, its purity and integrity. Fresh material, material stored at 2-8°C for 1-4 weeks and frozen at -80°C were tested...
March 9, 2017: Preparative Biochemistry & Biotechnology
https://www.readbyqxmd.com/read/28277818/extraction-of-rapamycin-sirolimus-from-streptomyces-rapamycinicus-using-ultrasound
#7
Amol S More, Sagar Gadalkar, Virendra K Rathod
The study was designed to investigate the use of Ultrasound to assisted extraction (UAE) of Rapamycin (sirolimus) from bacterial strain of Streptomyces rapamycinicus NRRL 5491. To achieve the maximum extraction yield, various parameters were optimized which includes Streptomyces rapamycinicus(10 g) of biomass in toluene (50 mL), temperature (20°C), electrical acoustic intensity (35.67 W/cm(2)) and duty cycle (40%) for 4 min with probe tip length of 0.5 cm dipped in to extraction solvent from the surface...
March 9, 2017: Preparative Biochemistry & Biotechnology
https://www.readbyqxmd.com/read/28267413/high-production-of-xanthan-gum-by-a-glycerol-tolerant-strain-xanthomonas-campestris-wxlb-006
#8
Zichao Wang, Jianrong Wu, Min-Jie Gao, Li Zhu, Xiao-Bei Zhan
The superior properties of xanthan gum make it an industrial aginomoto used in many industries, especially in oil recovery. In the present work, xanthan production from glycerol by a mutant strain Xanthomonas campestris WXLB-006 reached as high as 17.8 g/L in flask culture. With the adoption of pH control, varied aeration and agitation, and varied glycerol feeding strategy, xanthan production reached 33.9 g/L in a 7-L fermenter and fermentation time decreased to 60 hr. Instead of difficultly and costly purifying glycerol, this research provides a very good case for glycerol utilization...
February 21, 2017: Preparative Biochemistry & Biotechnology
https://www.readbyqxmd.com/read/28278108/influence-of-al-3-on-the-titer-of-spiramycin-and-effective-components-in-fermentor
#9
Zhen Li, Fengxian Hu, Ruifang Ye, Heping Lv, Jun Zeng
Spiramycin is a multi-component antibiotic and different components have different antibacterial activities. In Streptomyces spiramyceticus16-10-2, SPMII and SPMIII (spiramycin II and spiramycin III) are the main components, while SPMI (spiramycin I) need to be controlled below 12%. Based on this, the influences of Al(3+) on total spiramycin titer and components were investigated in this work. Those experiments were mainly performed in 15 L fermentor and Al(3+) made a great improvement in spiramycin titer...
February 17, 2017: Preparative Biochemistry & Biotechnology
https://www.readbyqxmd.com/read/28278109/identification-and-production-of-bioflocculants-by-enterobacter-sp-and-bacillus-sp-and-their-characterization-studies
#10
L Muthulakshmi, H Nellaiah, T Kathiresan, N Rajini, Fenila Christopher
In this work, two bioflocculants, namely, EB-EPS and B1-EPS were derived from Enterobacter sp. and Bacillus sp., respectively and analyzed with regard to their production and characterization. About 0.9 g and 0.16 g of purified EB and B1 were obtained from I L of fermentation broth. Chemical analysis showed the contents of purified EB and B1 mainly as 88.7% and 92.8% (w/w) of carbohydrate, and 11.3% and 21.8% (w/w) protein, respectively. Fourier-transform infrared (FTIR) spectrometry analysis revealed the presence of hydroxyl, amide and carboxyl groups in the identified bioflocculant...
February 16, 2017: Preparative Biochemistry & Biotechnology
https://www.readbyqxmd.com/read/28278112/purification-and-characterization-of-a-novel-trypsin-like-protease-from-green-seeded-chickpea-cicer-arientum
#11
Tooba Naz Shamsi, Romana Parveen, Priyankar Sen, Sadaf Fatima
The present study describes the purification and physicochemical and biochemical characterization of trypsin-like protease from green-seeded chickpea (Cicer arientum). The crude extract of chickpea trypsin (CpT) was obtained by homogenization followed by differential ammonium sulfate precipitation. The CpT was purified by ion-exchange chromatography on diethylaminoethyl (DEAE) column, pre-equilibrated with 20 mM tris-CaCl2 buffer (pH 8.2) with a flow rate of 0.5 mL min(-1). The molecular weight and purity of ∼23 kDa of CpT were determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis...
February 10, 2017: Preparative Biochemistry & Biotechnology
https://www.readbyqxmd.com/read/28278111/box-behnken-analysis-and-storage-of-spray-dried-collagenolytic-proteases-from-myceliophthora-thermophila-submerged-bioprocess
#12
Youssef Ali Abou Hamin Neto, Luciana Barbosa Coitinho, Luis Alexandre Pedro de Freitas, Hamilton Cabral
Enzymes do not have long-term storage stability in soluble forms, thus drying methods could minimize the loss of enzymatic activity, the spray dryer removes water under high temperatures and little time. The aims of this study were to improve the stability of enzymatic extract from Myceliophthora thermophila for potential applications in industry and to evaluate the best conditions to remove the water by spray drying technique. The parameters were tested according to Box-Behnken and evaluated by analysis of variance (ANOVA), all the parameters measured were found to influence the final enzyme activity and spray drying process yield ranged from 38...
February 10, 2017: Preparative Biochemistry & Biotechnology
https://www.readbyqxmd.com/read/28278110/optimization-of-penicillium-aurantiogriseum-protease-immobilization-on-magnetic-nanoparticles-for-antioxidant-peptides-obtainment
#13
José Manoel Wanderley Duarte Neto, Jackeline da Costa Maciel, Júlia Furtado Campos, Luiz Bezerra de Carvalho Junior, Daniela Araújo Viana Marques, Carolina de Albuquerque Lima, Ana Lúcia Figueiredo Porto
This work reports an optimization of protease from Penicillium aurantiogriseum immobilization on polyaniline coated magnetic nanoparticles for antioxidant peptides obtainment derived from bovine casein. Immobilization process was optimized using a full two-level factorial design (2(4)) followed by a response surface methodology (RSM). Using the derivative, casein was hydrolyzed uncovering its peptides that were sequenced and had antioxidant properties tested via ABTS radical scavenging and Hydrogen peroxide scavenging assays...
February 10, 2017: Preparative Biochemistry & Biotechnology
https://www.readbyqxmd.com/read/28165864/comprehensive-utilization-of-activated-sludge-for-preparation-of-hydrolytic-enzymes-polyhydroxyalkanoates-and-water-retaining-organic-fertilizer
#14
He Ni, Xiao-Min Fan, Hao-Ning Guo, Jian-Hua Liang, Qing-Rong Li, Liu Yang, Hui Li, Hai-Hang Li
The urban wastewater treatment industry produces a large amount of excess activated sludge which is mainly composed of microbial biomass and costly to be disposed. In this research, a comprehensive utilization of activated sludge was developed by sequentially extracting hydrolytic enzymes and polyhydroxyalkanoates (PHAs), and the residue was used to prepare water-retaining organic fertilizer. The sludge was extracted with 4-fold H2O containing 1% Triton X-100 with the yield of 66.7% protease activity. The enzyme solution was precipitated in 80% acetone and vacuum-dried at 40°C at the dried enzyme yield of 2...
February 6, 2017: Preparative Biochemistry & Biotechnology
https://www.readbyqxmd.com/read/28151056/%C3%AE-decalactone-production-by-yarrowia-lipolytica-and-lindnera-saturnus-in-crude-glycerol
#15
Géssyca P A Soares, Karla S T Souza, Leonardo F Vilela, Rosane F Schwan, Disney R Dias
Flavor compounds are commonly obtained from chemical synthesis or extracted from plants. These sources have disadvantages, such as racemic mixtures generation, more steps to yield the final product, low yield, and high cost, making the microbial fermentation an alternative and potential way to obtain flavor compounds. The most important lactone for flavor application is γ-decalactone, which has the aroma of peach, and can be obtained by ricinoleic acid biotransformation via yeast peroxisomal β-oxidation. The aim of this work was to use crude glycerol, a residual biodiesel industry, for the production of bioaroma from two different yeasts...
February 2, 2017: Preparative Biochemistry & Biotechnology
https://www.readbyqxmd.com/read/28151045/hirudin-as-a-novel-fusion-tag-for-efficient-production-of-lunasin-in-escherichia-coli
#16
Qinghua Tian, Ping Zhang, Zhan Gao, Hengli Li, Zhengli Bai, Shuhua Tan
Fusion expression provides an effective means for the biosynthesis of longer peptides in Escherichia coli. However, the commonly used fusion tags are primarily suitable for laboratory scale applications due to the high cost of the commercial affinity resins. Herein, a novel approach exploiting hirudin as a multipurpose fusion tag in combination with TEV protease cleavage has been developed for the efficient and cost-effective production of a 43-amino acids model peptide lunasin in E. coli at preparative scale...
February 2, 2017: Preparative Biochemistry & Biotechnology
https://www.readbyqxmd.com/read/28140750/production-of-a-protease-inhibitor-from-edible-mushroom-agaricus-bisporus-and-its-statistical-optimization-by-response-surface-methodology
#17
Reena Vishvakarma, Abha Mishra
The production of a protease inhibitor from Agaricus bisporus through solid-state fermentation was studied. The purpose was to produce protease inhibitor from natural, cheap, and readily available carbon and nitrogen sources. Solid-state fermentation enhanced the mycelia growth and also gave a higher yield of the product. Further, fungal growth and other production parameters were statistically optimized. The specificity of the inhibitor was tested and was effective against trypsin. Screening of significant factors (wheat bran, cyanobacterial biomass, initial pH, temperature, incubation period, and moisture content and inoculum size) was performed using Plackett-Burman design...
January 31, 2017: Preparative Biochemistry & Biotechnology
https://www.readbyqxmd.com/read/28106512/endoglucanase-and-xylanase-production-by-bacillus-sp-ar03-in-co-culture-glycosyl-hydrolases-by-bacillus-sp-ar03-in-co-culture
#18
Johan S Hero, José H Pisa, Nora I Perotti, Cintia M Romero, María A Martínez
The behaviour of three isolates retrieved from different cellulolytic consortia, Bacillus sp. AR03, Paenibacillus sp. AR247 and Achromobacter sp. AR476-2, were examined individually and as co-cultures in order evaluate their ability to produce extracellular cellulases and xylanases. Utilizing a peptone-based medium supplemented with carboxymethyl cellulose (CMC), an increase estimated of 1.30 and 1.50 times was obtained by the co-culture containing the strains AR03 and AR247, with respect to enzyme titles registered by their individual cultivation...
January 20, 2017: Preparative Biochemistry & Biotechnology
https://www.readbyqxmd.com/read/28102768/an-experimental-strategy-validated-to-design-cost-effective-culture-media-based-on-response-surface-methodology
#19
J L Navarrete-Bolaños, G P Téllez-Martínez, R Miranda-López, H Jiménez-Islas
For any fermentation process, the production cost depends on several factors, such as the genetics of the microorganism, the process condition and the culture medium composition. In this work, a guideline for the design of cost-efficient culture media using a sequential approach based on response surface methodology is described. The procedure was applied to analyze and optimize a culture medium of registered trademark and a base culture medium obtained as a results of the screening analysis from different culture media used to grow the same strain according to the literature...
January 19, 2017: Preparative Biochemistry & Biotechnology
https://www.readbyqxmd.com/read/28045643/production-of-human-mutant-biologically-active-hepatocyte-growth-factor-in-chinese-hamster-ovary-cells
#20
Dongsheng Xu, Aini Wan, Lin Peng, Yun Chen, Yang He, Jianfeng Yang, Jian Jin
Hepatocyte growth factor (HGF) is a potent multifunctional cytokine that affects proliferation, migration, and morphogenesis of various cells. HGF is secreted as an inactive single-chain precursor protein and activated by the cleavage of serine proteases to form heterodimers. In our current study, the cleavage site of HGF was blocked by replaced Arg 494 of Glu (R494E) that resulted in the single-chain HGF (R494E) unable to be cleaved by serine proteases. We established Chinese hamster ovary (CHO) cells overexpressing HGF (R494E), the expression of HGF (R494E) achieved 12 mg/L and was similar to a previously reported study...
January 3, 2017: Preparative Biochemistry & Biotechnology
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