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Preparative Biochemistry & Biotechnology

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 (PBD) was implemented to screen for the key medium components for the PQQ production. CoCl2 · 6H2O, ρ-amino benzoic acid (PABA), and MgSO4 · 7H2O were found capable of enhancing the PQQ production most significantly. A five level-three factor central composite design (CCD) was employed to investigate the direct and interactive effects of these variables...
April 27, 2017: Preparative Biochemistry & Biotechnology
Arunangshu Das, Anita Verma, Krishna J Mukherjee
L-Dopa and dopamine are important pathway intermediates towards the synthesis of catecholamine like 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 E.coli DH5α by 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
Shima Zamanian, Samira Mohammadi-Yeganeh, Vahid Kia, Hooman Kaghazian, Mahdi Paryan
During the purification of Human Recombinant EPO (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 (TSG) 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 picograms/dose. In this study we have designed a sensitive and specific Quantitative Real-time PCR assay for detection of residual DNA in human rEPO products...
April 20, 2017: Preparative Biochemistry & Biotechnology
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 rised to 100 g/L, the acetoin yield reached 24...
April 20, 2017: Preparative Biochemistry & Biotechnology
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 PBD revealed that the silk cocoon and PLA film were the most significant variables enhancing the PLA-degrading enzyme production. After a RSM, a maximum amount of PLA-degrading enzyme activity at 0.74 U mL(-1) was predicted and successfully validated at 95% after 0.39% (w/v) silk cocoon and 1...
April 17, 2017: Preparative Biochemistry & Biotechnology
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
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
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-polyethylene glycol (PEG) method, to obtain functional IgY with high purity and yield...
April 12, 2017: Preparative Biochemistry & Biotechnology
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
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
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
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
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
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
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
Amol S More, Sagar Gadalkar, Virendra K Rathod
The study was designed to investigate the use of ultrasound-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 include S. rapamycinicus (10 g) of biomass in toluene (50 mL), temperature (20°C), acoustic intensity (35.67 W/cm(2)), and duty cycle (40%) for 4 min extraction time with probe tip length of 0.5 cm dipped into extraction solvent from the surface...
March 9, 2017: Preparative Biochemistry & Biotechnology
Sepideh Khazeni, Ashrafalsadat Hatamian-Zarmi, Fatemeh Yazdian, Zahra Beagom Mokhtari-Hosseini, Bahman Ebrahimi-Hosseinzadeh, Behnam Noorani, Ghassem Amoabedini, Mohammad Reza Soudi
Bacterial cellulose (BC) is a very fascinating microbial biopolymer which is mainly produced by Gluconacetobacter xylinum. Optimization of BC production by G. xylinum was performed based on scale-down studies in miniature-bioreactor and response surface methodology in which the optimum pH value (6.5) and shaking rate (50 rpm) were obtained. The static culture condition for BC production has newly been defined. Nanostructure of BC includes nanofibers up to (60 nm) and nanoporosity up to (265 nm) was observed by scanning electron microscopy...
April 21, 2017: Preparative Biochemistry & Biotechnology
Hanen Mallek-Fakhfakh, Jawhar Fakhfakh, Najla Masmoudi, Fatma Rezgui, Ali Gargouri, Hafedh Belghith
In the present study, we investigated a potent extracellular β-glucosidases secreted by the thermophilic fungal strain AX4 of Talaromyces thermophilus, isolated from Tunisian soil samples. This strain was selected referring to the highest thermostability of its β-glucosidases compared to the other fungal isolates. The β-glucosidase production was investigated by submerged fermentation. The optimal temperature and initial pH for maximum β-glucosidase production were 50°C and 7.0, respectively. Several carbon sources were assayed for their effects on β-glucosidase production, significant yields were obtained in media containing lactose 1% (3...
April 21, 2017: Preparative Biochemistry & Biotechnology
Siriporn Chaikaew, Apinun Kanpiengjai, Jenjira Intatep, Kridsada Unban, Pairote Wongputtisin, Goro Takata, Chartchai Khanongnuch
The present study demonstrates the effectiveness of X-ray radiation in strain improvement for defective lipase production by Bacillus sp. MR10 for further application in the fermentative production of manno-oligosaccharides (MOS) from agricultural by-product, defatted copra meal (DCM). The mutants obtained were screened based on their defective lipase activity together with their β-mannanase production performance. Among 10 selected mutants, the strain M7 was the highest promising mutant regarding the smallest lipase activity (0...
April 21, 2017: Preparative Biochemistry & Biotechnology
Ashish A Prabhu, A Jayadeep
The current study is focused on optimizing the parameters involved in enzymatic processing of red rice bran for maximizing total polyphenol (TP) and free radical scavenging activity (FRSA). The sequential optimization strategies using central composite design (CCD) and artificial neural network (ANN) modeling linked with genetic algorithm (GA) was performed to study the effect of incubation time (60-90 min), xylanase concentration (5-10 mg/g), cellulase concentration (5-10 mg/g) on the response, i.e., total polyphenol and FRSA...
April 21, 2017: Preparative Biochemistry & Biotechnology
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