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Preparative Biochemistry & Biotechnology

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https://www.readbyqxmd.com/read/28644760/expression-and-characterization-of-soybean-seed-coat-peroxidase-in-escherichia-coli-bl21-de3
#1
Changqing Liu, Kai Zheng, Ying Xu, Lacmata Tamekou Stephen, Jiming Wang, Hongwei Zhao, Tongqing Yue, Rui Nian, Haibo Zhang, Mo Xian, Huizhou Liu
Soybean seed coat peroxidase(SBP) is a valuable enzyme having a broad variety of applications in analytical chemistry, biochemistry and food processing. In the present study, the sscp gene (Gene ID: 548068) was optimized based on the preferred codon usage of Escherichia coli, synthesized, and expressed in E. coli BL21(DE3). SDS-PAGE and western blot analysis of this expressed protein revealed that its molecular weight is approximately 39 kDa. The effects of induction temperature, concentration of IPTG (isopropyl-β-D-thiogalactoside) and hemin, induction time, expression time were optimized to enhance SBP production with a maximum activity of 11...
June 23, 2017: Preparative Biochemistry & Biotechnology
https://www.readbyqxmd.com/read/28644710/study-of-artemisinin-and-sugars-accumulation-in-artemisia-vulgaris-and-artemisia-dracunculus-hairy-root-cultures
#2
Kateryna O Drobot, Nadiia A Matvieieva, Andriy M Ostapchuk, Maxim A Kharkhota
We studied the effect of genetic transformation on biologically active compounds (artemisinin and its co-products (ART) as well as sugars) accumulation in Artemisia vulgaris and A.dracunculus "hairy" root cultures. Glucose, fructose, sucrose and mannitol were accumulated in A. vulgaris and A. dracunculus "hairy" root lines. Genetic transformation has led in some cases to the sugars content increasing or appearing of non-relevant for the control plants carbohydrates. Sucrose content was 1.6 times higher in A...
June 23, 2017: Preparative Biochemistry & Biotechnology
https://www.readbyqxmd.com/read/28636483/lumping-kinetics-of-abe-fermentation-wastewater-treatment-by-oleaginous-yeast-trichosporon-cutaneum
#3
Can Wang, Chao Huang, Xue-Fang Chen, Hai-Rong Zhang, Lian Xiong, Xiao-Mei Li, Hai-Jun Guo, Gao-Xiang Qi, Xiao-Qing Lin, Xin-De Chen
Lumping kinetics models were built for the biological treatment of ABE (acetone-butanol-ethanol) fermentation wastewater by oleaginous yeast Trichosporon cutaneum with different fermentation temperatures. Compared with high temperature (33°C, 306 K) and low temperature (23°C, 296 K), medium temperature (28°C, 301 K) was beneficial for the cell growth and COD degradation during the early stage of fermentation but the final yeast biomass and COD removal were influenced little. By lumping method, the materials in the bioconversion network were divided into five lumps (COD, lipid, polysaccharide, other intracellular products, other extracellular products), and the nine rate constants (k1-k9) for the models can well explain the bioconversion laws...
June 21, 2017: Preparative Biochemistry & Biotechnology
https://www.readbyqxmd.com/read/28636478/pathway-and-rate-limiting-step-of-glyphosate-degradation-by-aspergillus-oryzae-a-f02
#4
Gui-Ming Fu, Yan Chen, Ru-Yi Li, Xiao-Qiang Yuan, Cheng-Mei Liu, Bin Li, Yin Wan
Aspergillus oryzae A-F02, a glyphosate-degrading fungus, was isolated from an aeration tank in a pesticide factory. The pathway and rate-limiting step of glyphosate (GP) degradation were investigated through metabolite analysis. GP, aminomethylphosphonic acid (AMPA) and methylamine were detected in the fermentation liquid of A. oryzae A-F02, whereas sarcosine and glycine were not. The pathway of GP degradation in A. oryzae A-F02 was revealed: GP was firstly degraded into AMPA, which was then degraded into methylamine...
June 21, 2017: Preparative Biochemistry & Biotechnology
https://www.readbyqxmd.com/read/28636463/optimization-of-an-anti-her2-nanobody-expression-using-the-taguchi-method
#5
Alireza Farasat, Fatemeh Rahbarizadeh, Davoud Ahmadvand, Fatemeh Yazdian
Despite being widely used in immunotherapy of cancer, whole antibodies are limited by several disadvantages. This has led to the advent of novel biomolecules such as nanobodies. Taguchi method is a statistical experimental design to study the effect of multiple variables in biological processes. In an effort to overexpress a recombinant anti-HER2 nanobody, we carried out a detailed study to find optimal condition of temperature, induction, culture media, vector and host strain, using Taguchi methodology. A total of 16 various experiments were designed...
June 21, 2017: Preparative Biochemistry & Biotechnology
https://www.readbyqxmd.com/read/28636439/functional-enrichment-of-mannanase-treated-spent-brewer-yeast
#6
Ruge Cao, Xingyue Yang, Wenting Shang, Zhongkai Zhou, Padraig Strappe, Chris Blanchard
In this study, the effect of Bacillus brevis produced β-mannanase on the nutrients diffusion (release) and antioxidant activity of spent brewer yeast (SBY) was investigated. Three pre-treatments were performed: (1) autolysis at 50°C for 24 h; (2) autolysis at 50°C for 24 h, with the addition of β-mannanase during the autolysis; (3) autolysis at 50°C for 24 h, and the β-mannanase was added for another 12 h treatment. The pre-treatments with the addition of β-mannanase caused significant cell wall degradation; markedly increased the yield of SBY extracts...
June 21, 2017: Preparative Biochemistry & Biotechnology
https://www.readbyqxmd.com/read/28636436/signal-on-fluorescent-sensor-based-on-n-cqds-for-the-detection-of-glutathione-in-human-serum-and-pharmaceutic-preparation
#7
Chengfei Zhao, Kangying Wang, Qihong Cai, Haijian Tu, Linghong Pan, Liumin Yu
Nitrogen-doped carbon quantum dots (N-CQDs) with citric acid and ethylenediamine as raw materials were synthesized by an efficient one-step strategy. The N-CQDs showed a special property that the fluorescence was quenched by Fe(3+). The quenched fluorescence of N-CQDs could be recovered by glutathione (GSH). Therefore, a "signal-on" fluorescent sensor was developed to detect GSH. The fluorescent sensor could favorably avoid the interference of ascorbic acid, dopamine, glucose, oxidized glutathione and other amino acids in the detecting process of GSH...
June 21, 2017: Preparative Biochemistry & Biotechnology
https://www.readbyqxmd.com/read/28636431/enhanced-production-of-raw-starch-degrading-enzyme-using-agro-industrial-waste-mixtures-by-thermotolerant-rhizopus-microsporus-for-raw-cassava-chip-saccharification-in-ethanol-production
#8
Srisakul Trakarnpaiboon, Nantana Srisuk, Kuakoon Piyachomkwan, Kenji Sakai, Vichien Kitpreechavanich
In the present study, solid state fermentation for the production of raw starch degrading enzyme was investigated by thermotolerant Rhizopus microsporus TISTR 3531 using a combination of agro-industrial wastes as substrates. The obtained crude enzyme was applied for hydrolysis of raw cassava starch and chips at low temperature and subjected to non-sterile ethanol production using raw cassava chips. The agro-industrial wastes ratio was optimized using a simplex axial mixture design. The results showed that the substrate mixture consisting of rice bran: corncob: cassava bagasse at 8 g: 10 g: 2 g yielded the highest enzyme production of 201...
June 21, 2017: Preparative Biochemistry & Biotechnology
https://www.readbyqxmd.com/read/28635559/purification-of-antibody-against-ara-h-2-by-a-home-made-immunoaffinity-chromatography-column
#9
Zhihua Wu, Kun Li, Shaode Zhan, Ping Tong, Xin Li, Anshu Yang, Hongbing Chen
Antibodies are used extensively in numerous applications both in vivo and in vitro. To purify anti-Ara h 2 polyclonal antibody (PcaAb), a home-made immunoaffinity chromatography column (IAC) method was established. The properties of the home-made column were compared with those of the Mab Affinity Protein G Agarose High Flow (MPG), a commercially available column successfully used in capturing polyclonal antibodies. During antibody purification from rabbits antiserum against Ara h 2, the column capacity, recovery, and purification factor were characterized for IAC and MPG...
June 21, 2017: Preparative Biochemistry & Biotechnology
https://www.readbyqxmd.com/read/28635367/in-vivo-and-in-vitro-depolymerizations-of-intracellular-medium-chain-length-poly-3-hydroxyalkanoates-produced-by-pseudomonas-putida-bet001
#10
Siti Nor Syairah Anis, Mohamad Suffian Mohamad Annuar, Khanom Simarani
In vivo and in vitro depolymerizations of intracellular medium-chain-length poly-3-hydroxyalkanoates (mcl-PHA) in Pseudomonas putida Bet001 grown on lauric acid was studied. Both processes were studied under optimum conditions for mcl-PHA depolymerization viz. 0.2 M Tris-HCl buffer, pH 9, ionic strength (I) = 0.2 M at 30 °C. For in vitro depolymerization studies, cell-free system was obtained from lysing bacterial cells suspension by ultrasonication at optimum conditions (frequency 37 kHz, 30% of power output, <25 (°)C for 120 minutes)...
June 21, 2017: Preparative Biochemistry & Biotechnology
https://www.readbyqxmd.com/read/28448745/medium-optimization-for-pyrroloquinoline-quinone-pqq-production-by-methylobacillus-sp-zju323-using-response-surface-methodology-and-artificial-neural-network-genetic-algorithm
#11
Peilian Wei, Zhenjun Si, Yao Lu, Qingfei Yu, Lei Huang, Zhinan Xu
Methylobacillus sp. zju323 was adopted to improve the biosynthesis of pyrroloquinoline quinone (PQQ) by systematic optimization of the fermentation medium. The Plackett-Burman design was implemented to screen for the key medium components for the PQQ production. CoCl2 · 6H2O, ρ-amino benzoic acid, and MgSO4 · 7H2O were found capable of enhancing the PQQ production most significantly. A five-level three-factor central composite design was used to investigate the direct and interactive effects of these variables...
April 27, 2017: Preparative Biochemistry & Biotechnology
https://www.readbyqxmd.com/read/28430025/synthesis-of-dopamine-in-e-coli-using-plasmid-based-expression-system-and-its-marked-effect-on-host-growth-profiles
#12
Arunangshu Das, Anita Verma, Krishna J Mukherjee
L-Dopa and dopamine are important pathway intermediates toward the synthesis of catecholamine such as epinephrine and norepinephrine from amino acid L-tyrosine. Dopamine, secreted from dopaminergic nerve cells, serves as an important neurotransmitter. We report the synthesis of dopamine by extending the aromatic amino acid pathway of Escherichia coli DH5α by the expression of 4-hydroxyphenylacetate-3-hydrolase (HpaBC) from E. coli and an engineered dopa decarboxylase (DDC) from pig kidney cell. The activity of HpaBC and DDC require 200 µM iron supplementation and 50 µM vitamin B6, respectively as additives to the growth media...
April 21, 2017: Preparative Biochemistry & Biotechnology
https://www.readbyqxmd.com/read/28426392/design-and-validation-of-a-new-method-to-detect-and-quantify-residual-host-cell-dna-in-human-recombinant-erythropoietin-repo
#13
Shima Zamanian, Samira Mohammadi-Yeganeh, Vahid Kia, Hooman Kaghazian, Mahdi Paryan
During the purification of human recombinant erythropoietin (rEPO) from host cells, residual DNA may remain in final products. This contamination is a risk factor for patients and may result in the inactivation of some tumor suppressor genes or activation of oncogenes if its concentration is more than the standard defined by WHO. Based on WHO's criteria, acceptable level of residual DNA in biopharmaceuticals is less than 10-100 pg/dose. In this study, we have designed a sensitive and specific quantitative real-time polymerase chain reaction (PCR) assay for the detection of residual DNA in human rEPO products...
April 20, 2017: Preparative Biochemistry & Biotechnology
https://www.readbyqxmd.com/read/28426331/effect-of-deletion-of-2-3-butanediol-dehydrogenase-gene-bdha-on-acetoin-production-of-bacillus-subtilis
#14
Junjiao Zhang, Xiangying Zhao, Jiaxiang Zhang, Chen Zhao, Jianjun Liu, Yanjun Tian, Liping Yang
The present work aims to block 2,3-butanediol synthesis in acetoin fermentation of Bacillus subtilis. First, we constructed a recombinant strain BS168D by deleting the 2,3-butanediol dehydrogenase gene bdhA of the B. subtilis168, and there was almost no 2,3-butanediol production in 20 g/L of glucose media. The acetoin yield of BS168D reached 6.61 g/L, which was about 1.5 times higher than that of the control B. subtilis168 (4.47 g/L). Then, when the glucose concentration was increased to 100 g/L, the acetoin yield reached 24...
April 20, 2017: Preparative Biochemistry & Biotechnology
https://www.readbyqxmd.com/read/28414263/effective-enhancement-of-polylactic-acid-degrading-enzyme-production-by-amycolatopsis-sp-strain-scm_mk2-4-using-statistical-and-one-factor-at-a-time-approaches
#15
Watsana Penkhrue, Apinun Kanpiengjai, Chartchai Khanongnuch, Kazuo Masaki, Wasu Pathom-Aree, Winita Punyodom, Saisamorn Lumyong
This study aims to find the optimal medium and conditions for polylactic acid (PLA)-degrading enzyme production by Amycolatopsis sp. SCM_MK2-4. Screening of the most effective components in the enzyme production medium by Plackett-Burman design revealed that the silk cocoon and PLA film were the most significant variables enhancing the PLA-degrading enzyme production. After an response surface methodology, a maximum amount of PLA-degrading enzyme activity at 0.74 U mL(-1) was predicted and successfully validated at 95% after 0...
April 17, 2017: Preparative Biochemistry & Biotechnology
https://www.readbyqxmd.com/read/28409700/high-yield-of-biologically-active-recombinant-human-fibroblast-growth-factor-16-in-e-coli-and-its-mechanism-of-proliferation-in-ncl-h460-cells
#16
Jiliang Cheng, Zhaoxiang Fang, Huanhuan Yang, Yong Li, Haishan Tian, Weiyue Gong, Taotao Chen, Min Liu, Xiaokun Li, Chao Jiang
Fibroblast growth factor-16 (FGF16) is a member of FGF9 subfamily, which plays key role in promoting mitosis and cell survival, and also involved in embryonic development, cell growth, tissue repair, morphogenesis, tumor growth, and invasion. However, the successful high-yield purification of recombinant human fibroblast growth factor-16 (rhFGF16) protein has not been reported. In addition, lung cancer is a major cause of cancer-related deaths, which threats people's lives and its incidence has continued to rise...
April 14, 2017: Preparative Biochemistry & Biotechnology
https://www.readbyqxmd.com/read/28402220/implementation-of-quality-by-design-toward-processing-of-food-products
#17
Anurag S Rathore, Gautam Kapoor
Quality by design (QbD) is a systematic approach that begins with predefined objectives and emphasizes product and process understanding and process control. It is an approach based on principles of sound science and quality risk management. As the food processing industry continues to embrace the idea of in-line, online, and/or at-line sensors and real-time characterization for process monitoring and control, the existing gaps with regard to our ability to monitor multiple parameters/variables associated with the manufacturing process will be alleviated over time...
April 12, 2017: Preparative Biochemistry & Biotechnology
https://www.readbyqxmd.com/read/28402216/a-novel-and-efficient-immunoglobulin-y-extraction-method-using-poloxamer-polyethylene-glycol
#18
Cui Li, Hao Ren, Rüdiger Schade, Xiaoying Zhang
Although many IgY extraction methods (such as polyethylene glycol (PEG) precipitation method, octanoic acid method, water dilution method, etc.) have been established, there is still industrial drive and real need in developing scale-up IgY production methods. Some previous studies have reported that poloxamer degreasing method shows very good result in IgY extraction from egg yolk with high degreasing speed, harmlessness, simpleness in operation and minimal effect on antibody titer. In this study, we developed a new method, poloxamer-PEG method, to obtain functional IgY with high purity and yield...
April 12, 2017: Preparative Biochemistry & Biotechnology
https://www.readbyqxmd.com/read/28402172/rational-design-and-synthesis-of-affinity-matrices-based-on-proteases-immobilized-onto-cellulose-membranes
#19
Alberto Del Monte-Martínez, Jorge González-Bacerio, Bessy Cutiño-Avila, Jorge Rojas, Mae Chappé, Emir Salas-Sarduy, Isel Pascual, José M Guisán
Discovery of new protease inhibitors may result in potential therapeutic agents or useful biotechnological tools. Obtainment of these molecules from natural sources requires simple, economic and highly efficient purification protocols. The aim of this work was the obtainment of affinity matrices by the covalent immobilization of dipeptidyl peptidase IV (DPP-IV) and papain onto cellulose membranes, previously activated with formyl (FCM) or glyoxyl groups (GCM). GCM showed the highest activation grade (10.2 µmol aldehyde/cm(2))...
April 12, 2017: Preparative Biochemistry & Biotechnology
https://www.readbyqxmd.com/read/28267413/high-production-of-xanthan-gum-by-a-glycerol-tolerant-strain-xanthomonas-campestris-wxlb-006
#20
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...
May 28, 2017: Preparative Biochemistry & Biotechnology
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