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Preparative Biochemistry & Biotechnology

Yogesh Devaraj, Savita Kumari Rajender, Prakash Motiram Halami
A serine protease with preference for fibrin protein was purified and characterized from B.amyloliquefaciens MCC2606, isolated from dosa batter. The protease was purified using ammonium sulphate precipitation, ion-exchange and gel filtration chromatography. The degradation activity of the protease towards the fibrin was significantly higher compared with other protein substrates in the study. The molecular weight of the CFR15-protease was estimated to be 32 kDa based on SDS-PAGE. The purified enzyme exhibited both fibrinolytic and fibrinogenolytic activity...
January 17, 2018: Preparative Biochemistry & Biotechnology
Chong-Boon Ong, Mohamad S M Annuar
Immobilization of cross-linked tannase on pristine multi-walled carbon nanotubes (MWCNT) was successfully carried out. Cross-linking of tannase molecules was made via glutaraldehyde. The immobilized tannase exhibited significantly improved pH, thermal, and recycling stabilities. The optimal pH for both free- and immobilized tannase were observed at pH 5.0 with optimal operating temperature at 30 °C. Moreover, immobilized enzyme retained greater biocatalytic activities upon ten-repeated uses compared to free enzyme in solution...
January 17, 2018: Preparative Biochemistry & Biotechnology
Mona Liza Santana, Elinalva Maciel Paulo, José Ailton Bispo, Amanda Reges de Sena, Sandra Aparecida de Assis
The current study aims to assess the kinetics of population growth of R. oryzicola and the production of β-1,3-glucanase (EC enzyme by this yeast. It also aims to obtain the optimum conditions of β-1,3-glucanase enzymatic activity by varying the pH as well as to study the enzyme thermostability. R. oryzicola population doubled within 12 hours. During this period, 9.26 generations were obtained, with 1 hour and 29 minutes interval from one generation to the other, with specific growth rate (µ) of 0...
January 9, 2018: Preparative Biochemistry & Biotechnology
Jagruti Jadhav, Sruba Dutta, Sandeep Kale, Amit Prabhakar Pratap
Glycolipids are one of the major classes of biosurfactants in which the rhamnolipids are best studied. The present work investigates, optimization of inoculum age and batch time for maximizing the yield of rhamnolipid from Pseudomonas aeruginosa (MTCC 2453). The yield and titer of rhamnolipids were maximum in the fermentation batch with an inoculum age of 24 hours. Batch time studies were carried out on biomass production, rhamnolipid production and sunflower oil utilization. The maximum yield of rhamnolipid was achieved at 96 hours when the culture cells were in the late exponential/ early stationary phase...
January 9, 2018: Preparative Biochemistry & Biotechnology
Hadi Bayat, Saghar Hossienzadeh, Es'hagh Pourmaleki, Roshanak Ahani, Azam Rahimpour
Monoclonal antibodies (mAbs) have emerged as the most promising category of recombinant proteins due to their high efficiency for the treatment of a wide range of human diseases. The complex nature of mAbs creates a great deal of challenges in both upstream and downstream manufacturing processes. Proportional expression and correct folding and assembly of the light chain and heavy chain are required for efficient production of the mAbs. In this regard, expression vector design has proven to have profound effects on the antibody expression level as well as its stability and quality...
January 9, 2018: Preparative Biochemistry & Biotechnology
Tuğçe Yurtluk, Fikriye Alev Akçay, Ayşe Avcı
Biosynthesis of silver nanoparticles (AgNPs) using microorganisms is an important application of nano-biotechnology and green chemistry because of interest by pharmaceutical and food manufacturers. In this study, biosynthesis of AgNPs by a novel Bacillus strain isolated from a soil sample from Sakarya district in Turkey was investigated. Biosynthesis was performed using cell-free supernatant of the bacterium following 24 h growth. Effects of varying AgNO3 concentration (1-10 mM), pH (5-10), and temperature (30-40°C) on the synthesis of AgNPs were determined...
January 9, 2018: Preparative Biochemistry & Biotechnology
Zhongyu You, Suping Zhang, Xiaoxia Liu, Yujie Wang
PigC is a synthetase that catalyzes the condensation of 4-methoxy-2,2'-bipyrrole-5-carboxyaldehyde (MBC) and 2-methyl-3-amylpyrrole (MAP) to produce prodigiosin, which has a wide variety of impressive biological properties. In this study, we optimized PigC production from engineered Escherichia coli BL21(DE3). Investigation of different induction strategies revealed that auto-induction was the most appropriate method for PigC expression. As a result, PigC activity was elevated to 75.7 U/mL, nearly 2.1-fold higher than that with induction by isopropy-β-D-thiogalactoside (IPTG)...
January 9, 2018: Preparative Biochemistry & Biotechnology
Xiaofei Chen, Xiaoxiao Yu, Xiaoda Song, Li Liu, Yuting Yi, Wenbing Yao, Xiangdong Gao
Human epidermal growth factor receptor 2 (HER2) is a powerful target for cancer immune therapy. The development of anti-HER2 monoclonal antibodies targeting different domains of HER2 is quite effective. However, the selection and production of multivalent antibodies are complicated. In this study, a mimivirus-based designed ankyrin repeat protein (DARPin) targeting HER2 was selected from an artificial library by bacteria surface display. The selection was performed on HER2-positive B16BL6/E2 melanoma cells and HER2-nagative cells...
January 9, 2018: Preparative Biochemistry & Biotechnology
Mehri Mortazavi, Mohammad Ali Shokrgozar, Soroush Sardari, Kayhan Azadmanesh, Reza Mahdian, Hooman Kaghazian, Seyed Nezamedin Hosseini, Elnaz Shams
Recombinant protein aggregation is a problematic issue and can provoke immunological response. The aim of this study was to analyze the stability of erythropoietin, as a therapeutic protein expressed in mammalian cells, in the presence of different chemicals and find a specific stabilizer for erythropoietin. The effects of several chemicals, including mannitol, betaine, trehalose, taurine, linoleic acid, beta cyclodextrin, copper sulfate, spermidine, maltose, maltodextrin, sucrose, dextran, beta alanine, myo-inositol and cysteine, on protein stabilization through the thermally induced aggregation of erythropoietin were monitored...
January 9, 2018: Preparative Biochemistry & Biotechnology
E I Garcia-Peña, C Niño-Navarro, I Chairez, L Torres-Bustillos, J Ramírez-Muñoz, E Salgado-Manjarrez
In this study the biohydrogen (bioH2) production of a microbial consortium was optimized by adjusting the type and configuration of two impellers, the mixing regimen and the mass transfer process (KLa coefficients). A CSTR system, with a non-standard geometry, was characterized. Two different mixing configurations with either predominant axial (PB4 impeller) or radial pumping (Rushton impeller) were assessed and four different impeller configurations to produce bioH2. The best configuration for an adequate mixing time was determined by an ANOVA analysis...
December 7, 2017: Preparative Biochemistry & Biotechnology
Vibol Sok, Alex Fragoso
Carbon nanomaterials have emerged as suitable supports for enzyme immobilization and stabilization due to their inherently large surface area, high electrical conductivity, chemical stability and mechanical strength. In this paper, carbon nano-onions (CNOs) were used as supports to immobilize alkaline phosphatase, horseradish peroxidase and glucose oxidase. CNOs were first functionalized by oxidation to generate carboxylic groups on the surface followed by the covalent linking of the using a soluble carbodiimide as coupling agent...
December 7, 2017: Preparative Biochemistry & Biotechnology
Jian Zhang, Jie Yuan, Wen-Xue Zhang, Fang Tu, Ya Jiang, Chuan-Ze Sun
A novel and effective process was put forward for converting rice straw into feed by combining diluted acid hydrolysis and ammonization with Rhodospirillum rubrum fermentation. After pretreatment with dilute sulfuric or phosphoric acid (1%, w/w) at 100°C, materials were subjected to fermentation under several gases (N2, CO2, and air) and different light intensities in a 2-L fermentor. The key indexes of feed for fermented materials were estimated and several toxic substances were investigated during the fermentation...
December 1, 2017: Preparative Biochemistry & Biotechnology
Fatin Myra Abdul Manan, Nursyafreena Attan, Nashi Widodo, Hassan Y Aboul-Enein, Roswanira Abdul Wahab
An alternative environmentally benign support was prepared from chitosan-chitin nanowhiskers (CS/CNWs) for covalent immobilization of Rhizomucor miehei lipase (RML) to increase the operational stability and recyclability of RML in synthesizing eugenyl benzoate. The CS/CNWs support and RML-CS/CNWs were characterized using X-ray diffraction (XRD), fluorescent microscopy and Fourier transform infrared spectroscopy (FTIR). Efficiency of the RML-CS/CNWs was compared to the free RML to synthesize eugenyl benzoate for parameters: reaction temperature, stirring rate, reusability and thermal stability...
December 1, 2017: Preparative Biochemistry & Biotechnology
Preeti Tripathi, Jyothi Lakshmi A, Mukesh Kapoor
The present study was aimed at enhancing phytase (Phy-Ck) production from Citrobacter koseri PM-7 using response surface methodology (RSM) and improving the bioaccessibility of minerals (Fe and Zn) and protein digestibility in high-phytate food using Phy-Ck. A five-variable and three-level central composite design of RSM using wheat bran (6.681% w/v), inoculum level (2.5% v/v), and triton-x-100 (0.2% v/v) resulted in up to 5.57-fold (1.047 U/ml) improvement in Phy-Ck yield from C. koseri PM-7 when compared with fermentation media I & II...
December 1, 2017: Preparative Biochemistry & Biotechnology
S Lalitha Gavya, Neha Arora, Siddhartha Sankar Ghosh
A paradigm shift towards fusion proteins to render multiple functionalities and applications on a single platform has been incurred in enzyme based diagnosis. Herein, we report development and systematic characterizations of glutathione-s-transferase (GST) and lactate dehydrogenase-A (hLDHA) in a fusion protein (GST-hLDHA) to achieve functional activities of GST and hLDHA simultaneously. The GST-pGEX-4 T -2 vector system was employed for cloning and purification of human lactate dehydrogenase A, utilizing the affinity based interaction between GST and GSH in column chromatography...
December 1, 2017: Preparative Biochemistry & Biotechnology
Saima Nusrat, Rizwan Hasan Khan
The association of protein aggregates with plentiful human diseases has fascinated studies regarding the biophysical characterization of protein misfolding and ultimately their aggregate formation mechanism. Protein-ligand interaction, their mechanism, conformational changes by ligands and protein aggregate formation has been studied upon exploiting experimental techniques and computational methodologies. Such studies for the exploration of ligand induced conformational changes in protein, misfolding and aggregation, has confirmed drastic progresses in the study of aggregate formation pathways...
November 6, 2017: Preparative Biochemistry & Biotechnology
Manoj Kumar Narasimhan, Selvarajan Ethiraj, Tamilarasan Krishnamurthi, Mathur Rajesh
No abstract text is available yet for this article.
November 6, 2017: Preparative Biochemistry & Biotechnology
Guo-Li Gong, Yuan Wei, Zhong-Zhong Wang
In this study, a novel heterozygous antimicrobial peptide MLH was synthesized, expressed, purified and characterized. The peptide MLH was selected through bioinformatic analysis using musca domestica antimicrobial peptide (Cec-Med), human antimicrobial peptide LL-37 and helicobacter pylori antimicrobial peptide (Hp) as parent peptides. The target gene was synthesized by overlap extension PCR (SOE-PCR) and connected to the expression vector pET-32a (+), and the recombinant plasmid pET-32a-MLH was transformed to Escherichia coli for constructing pET-32a-MLH/BL21 (DE3)...
November 3, 2017: Preparative Biochemistry & Biotechnology
Zhen Rong, Ying-Yi Huo, Shu-Ling Jian, Yue-Hong Wu, Xue-Wei Xu
A novel esterase gene (e25) was identified from Altererythrobacter epoxidivorans CGMCC 1.7731(T) by genome sequence screening. The e25 gene is 948 nucleotides in length and encodes a 315 amino acid protein (E25) with a predicted molecular mass of 33,683 Da. A phylogenetic tree revealed that E25 belongs to the hormone-sensitive lipase (HSL) family of lipolytic enzymes. An activity assay of E25 showed that it exhibited the highest catalytic efficiency when using p-nitrophenyl caproate (C6) as a substrate. The optimum pH and temperature were determined to be approximately pH 9 and 45°C, and the Km and Vmax values were 0...
November 3, 2017: Preparative Biochemistry & Biotechnology
Ashish A Prabhu, Veeranki Venkata Dasu
Pichia pastoris is considered as one of the prominent host extensively used as a platform for heterologous protein production. In the present study, the growth inhibition kinetics of recombinant P. pastoris expressing human interferon gamma was studied under different initial substrate concentrations of gluconate (10-100 g L-1) and methanol (2-50 g L-1) in modified FM22 medium. The highest specific growth rate of 0.0206 and 0.019 hr-1 was observed at 60 g L-1 of gluconate and 10 g L-1 of methanol, respectively...
November 26, 2017: Preparative Biochemistry & Biotechnology
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