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Preparative Biochemistry & Biotechnology

Luane Ferreira Garcia, Monike Fabiane Alves Ribeiro Lacerda, Douglas Vieira Thomaz, Jhéssica Cavalcante de Souza Golveia, Maria das Graças Cabral Pereira, Eric de Souza Gil, Fernando Schimidt, Mariângela Fontes Santiago
Laccase extract (LE) from Pycnoporus sanguineus was immobilized on calcium and copper alginate-chitosan beads and applied for the removal of 17α-ethinylestradiol (EE2). Effects of immobilization conditions such as: sodium alginate (SA) concentration; LE/SA ratio and chitosan/ion (Ca+2 or Cu+2 ) ratio on the immobilization yield were investigated. Immobilized LE on Ca-beads and Cu-beads was then used to degrade an EE2 solution. The optimal conditions for LE immobilization on Ca-beads were: 1.5% (w/v) SA, 1:5 (v/v) LE/SA and 3:7 (v/v) chitosan/ion (Ca+2 )...
February 19, 2019: Preparative Biochemistry & Biotechnology
Mosayeb Zolfaghari-Esmaeelabadi, Parisa Hejazi
This research develops on our previous semi-mechanistic model that describes the dynamic physical and biochemical processes taking place in a packed-bed bioreactor to analyze the relationship of nutrient limitation, biomass accumulation, metabolic heat generation, and mathematical description of packed-bed porous media. The experimental and simulation data proved that glucose concentration gradients in the biofilm could be neglected due to small biofilm thickness and high diffusivity of glucose in the biofilm...
February 15, 2019: Preparative Biochemistry & Biotechnology
Herng C Leow, Katja Fischer, Yee C Leow, Katleen Braet, Qin Cheng, James McCarthy
Shark variable new antigen receptors (VNARs) are known to possess excellent heat-stability, and the long complementarity determining region 3 (CDR3) has permitted it to penetrate into the cleft region of antigens. The number of cysteine (Cys) residues contained within VNAR is greater than in conventional antibodies, entailing disulfide bond formation in both the inter- or intra-loop regions is required for interactions with the target protein antigens. Therefore, the selection of a suitable expression system is important to ensure the solubility and correct folding of functional VNAR protein production...
February 15, 2019: Preparative Biochemistry & Biotechnology
Hak Sup Shin, Soon-Ik Park
In this study, we report a novel member of the attacin family from Hermetia illucens. The cDNA clone encoding the attacin-like protein was isolated by screening a cDNA library prepared from immunized fat body. The complete 510 bp cDNA of HI-attacin was predicted to encode a protein of 169 amino acids with a molecular weight of 17.7 kDa. The putative mature protein of H. illucens attacin (HI-attacin) had 50% identity with that of Bactrocera dorsalis attacin B. Phylogenetic analysis revealed that the HI-attacin was separated from the other dipteran attacins with a high bootstrap percentage...
February 15, 2019: Preparative Biochemistry & Biotechnology
Ruiyan Zhu, Xiaojia Gao, Yan Xiao, Na Yang, Yangzhuoyue Jin
We conducted industrial scale γ-polyglutamic acid (γ-PGA) production by Bacillus subtilis (B. subtilis) LX and modeled its microbial growth kinetics based on a logistic regression. We found that the use of a three-layer impeller including a lower semicircular disc impeller and two-layers of six-wide-leaf impellers were able to both increase γ-PGA yields and decrease fermentation time as compared with two-layer Rushton impellers. Indeed, our results revealed that the optimal γ-PGA yield (20.67 ± 2.19 g/L) was obtained after 40 hr in the impeller retrofitted fermenter, and this yield was 29...
February 15, 2019: Preparative Biochemistry & Biotechnology
Yonca Yuzugullu Karakus, Semih Isik
Aqueous two-phase partitioning system (ATPS) was used to extract and purify catalase from Bacillus pumilus. The system parameters for effective purification of catalase were optimized. The best catalase recovery (123%) with a 4.6-fold purification was obtained in the bottom phase of ATPS including the mixture of 15% (w/w) PEG4000, 10% (w/w) Na2 SO4 and 3% (w/w) NaCl at pH 5.0. The purified enzyme was characterized regarding its activity and stability. The highest enzyme activity was observed at pH 7.0 and 37 °C on hydrogen peroxide...
February 15, 2019: Preparative Biochemistry & Biotechnology
Agnes Rodler, Rene Ueberbacher, Beate Beyer, Alois Jungbauer
Hydrophobic interaction chromatography is a very popular chromatography method for purification of proteins and plasmids in all scales from analytical to industrial manufacturing. Despite this frequent use, the complex interaction mechanism and the thermodynamic aspects of adsorption in hydrophobic interaction chromatography are still not well understood. Calorimetric methods such as isothermal titration calorimetry and flow calorimetry can help to gain a deeper understanding of the adsorption strength, the influence of salt type and temperature...
February 8, 2019: Preparative Biochemistry & Biotechnology
Samira Pourtaghi-Anvarian, Samin Mohammadi, Maryam Hamzeh-Mivehroud, Ali Akbar Alizadeh, Siavoush Dastmalchi
Single-chain fragment variable (scFv) antibodies are antibody fragments consist of variable domains of full antibodies known to retain antigen binding properties while having much lower molecular weights granting some beneficial properties to them. In our previous study, three phage particles each displaying an individual scFv antibody (i.e. J43, J44, and J48) were identified as tumor necrosis factor alpha (TNF-α) binders. The current study aimed to produce previously identified anti-TNF-α scFv antibodies and to investigate their abilities to bind and inhibit TNF-α biological effect...
February 8, 2019: Preparative Biochemistry & Biotechnology
Bahareh Azarian, Amin Azimi, Mahboubeh Sepehri, Vahideh Samimi Fam, Faegheh Rezaie, Yeganeh Talebkhan, Vahid Khalaj, Fatemeh Davami
Aggregation of recombinant proteins, a major problem in E. coli expression system, is improved by using EnBase culture system based on slow release of glucose. In the present study, to understand the intracellular mechanisms involved in increased solubility of the target recombinant protein through EnBase system, the effect of this system was investigated on E. coli cells proteome profile. The proteome profile of E. coli cells cultured in EnBase and conventional batch mode was analyzed by two-dimensional gel electrophoresis...
February 8, 2019: Preparative Biochemistry & Biotechnology
Deepali Verma, Monika Antil, Vibha Gupta
The emergence of drug resistance in Streptococcus pneumoniae (Spn) is a global health threat and necessitates discovery of novel therapeutics. The serine acetyltransferase (also known as CysE) is an enzyme of cysteine biosynthesis pathway and is reported to be essential for the survival of several pathogenic bacteria. Therefore, it appears to be a very attractive target for structure-function understanding and inhibitor design. This study describes the molecular cloning of cysE from Spn in the pET21c vector and efforts carried out for expression and purification of active recombinant CysE...
February 8, 2019: Preparative Biochemistry & Biotechnology
Dan Zhao, Yao Wang, Jin Na, Wenxiang Ping, Jingping Ge
Lactic acid bacteria (LAB) is an ideal mannanase source due to the bio-safety guarantee. LAB can heterogeneously express β-mannanase or be directly used as β-mannanase-producing strains. This research originally optimized the fermentation condition for β-mannanase produced by Lactobacillus casei HDS-01. The applicable potential of the crude enzyme in juice clarification was investigated. Two-factorial design screened out three factors, i.e., fermentation time (p = 0.0001), glucose (p = 0.0013), and initial pH (p = 0...
February 8, 2019: Preparative Biochemistry & Biotechnology
Jun Zhao, Hai-Yang Yu, Yu Zhao, Feng-Hua Li, Wei Zhou, Bin-Bin Xia, Zhi-Yuan He, Jason Chen, Guo-Tuo Jiang, Ming-Li Wang
In this paper, we report a soluble expression based on Escherichia coli and two-step purification of a novel thioredoxin-tagged chicken interferon-α fusion protein (Trx-rChIFN-α) by using pET32a(+) expression system. The mature ChIFN-α gene was amplified by Reverse transcriptase-polymerase chain reaction (RT-PCR) and subcloned into pET-32a (+) vector prior to transformation into Rosetta (DE3) competent cells. After IPTG induction, the recombinant fusion protein was expressed efficiently in the soluble fraction...
February 8, 2019: Preparative Biochemistry & Biotechnology
Yue Li, Jing Zhao, Chunshan Quan, Liming Jin, Yongbin Xu, Ming Chen
Acylase AiiO is a novel quorum quenching enzyme with a broad substrate spectrum of acyl-homoserine lactones (AHLs) and has promising prospects in pathogen control. In this work, acylase AiiO production by a recombinant E. coli strain and its characterization were investigated; the acylase powder was further prepared and evaluated for effectiveness. A strategy of auto-induction combined with temperature regulation was developed to improve AiiO production. For the soluble AiiO protein in the cells, maximum production of 214...
February 3, 2019: Preparative Biochemistry & Biotechnology
Haider M Zwain, Sue-Min Chang, Irvan Dahlan
Microbial content formed in bioreactors plays a significant role in the anaerobic process. Therefore, the physicochemical characteristics of microbial content in a modified anaerobic inclining-baffled reactor (MAI-BR) treating recycled paper mill effluent (RPME) were investigated using Fourier transform infrared (FTIR), X-ray diffraction (XRD), X-ray photoelectron spectroscopy (XPS), thermogravimetric (TG), and derivative thermogravimetric (DTG) analyses, scanning electron microscopy-energy dispersive X-ray spectroscopy (SEM-EDS), Brunauer-Emmett-Teller (BET), and surface area analyzer...
February 3, 2019: Preparative Biochemistry & Biotechnology
Santosh Kumar Karn, Awanish Kumar
Excess of sludge generated during activated sludge process by various industries is causing sludge disposable problem worldwide. Sludge contains organic materials of a highly complex mixture of molecules with some toxic compounds. Thus, the harmful effect of sludge could be managed by processing of sludge and recovery of useful products. Sludge could undergo biodegradation and used for the recovery of enzymes and energy. Enzymes (amylase, alkaline phosphatase, lipase, proteases) are produced by microorganisms and released into the media to degrade organic matters of sludge...
February 3, 2019: Preparative Biochemistry & Biotechnology
Nithya Vadakedath, Prakash M Halami
The antibacterial peptide of Bacillus licheniformis MCC 2016 have potential biopreservative efficacy. Here, we report the purification process, properties, and mode of action of this antibacterial peptide for its potential application in the food industry. The antibacterial peptide from the cell-free supernatant was purified using a sequence of purification steps. The purified antibacterial peptide showed a specific activity of 68817 AU mg-1 and 0.4% yield. Liquid chromatography-mass spectroscopy analysis showed an mz-1 value of 279...
February 3, 2019: Preparative Biochemistry & Biotechnology
Atefeh Ameri, Mojtaba Shakibaie, Mozhde Soleimani-Kermani, Mohammad Ali Faramarzi, Hamid Forootanfar
Microbial enzymes of extremophilic origin serve as a vital source of stable industrial enzymes. The present study focused on overproduction of a thermoalkalophilic lipase produced by Bacillus atrophaeus FSHM2 through UV-induced random mutagenesis (5-45 min exposure to UV light) and factorial experimental design augmented to response surface methodology. Firstly, a UV-induced mutant (designated as UV-45) was developed after the exposure of wild strain to UV irradiation for 45 min which was able to secrete 3484...
February 3, 2019: Preparative Biochemistry & Biotechnology
J J Muso Cachumba, R Terán Hilares, L P Brumano, P R F Marcelino, F A F Antunes, J C Santos, S S da Silva
L-asparaginase is an important enzyme used in the pharmaceutical and food industry, which can be produced by different microorganisms using low cost feedstocks. In this work, sugarcane bagasse (SCB) was used as support for enzyme production in solid-state fermentation (SSF) by A. terreus. Initially, the influence of the variables carbon and nitrogen sources on the enzyme production was studied following an experimental design carried out in Erlenmeyer flasks. Statistical analysis indicated the use of 0.54% of starch, 0% of maltose, 0...
February 3, 2019: Preparative Biochemistry & Biotechnology
Ahmad Beiroti, Mohammad Reza Aghasadeghi, Seyed Nezamedin Hosseini, Dariush Norouzian
Artificial neural networking (ANN) seems to be a promising soft sensor for implementing current approaches of quality by design (QbD) and process analytical technologies (PAT) in the biopharmaceutical industry. In this study, we aimed to implement best-fitted ANN architecture for online prediction of the biomass amount of recombinant Pichia pastoris (P. pastoris) - expressing intracellular hepatitis B surface antigen (HBsAg) - during the fed-batch fermentation process using methanol as a sole carbon source...
February 1, 2019: Preparative Biochemistry & Biotechnology
Duk-Un Kang, Yong-Suk Lee, Jin-Woo Lee
To enhance recovery yield of carboxymethylcellulase (CMCase), E. coli BL21/A-53 producing the histidine-tagged CMCase was constructed in this study. The recovery yield of the histidine-tagged CMCase using the His-tag affinity chromatography was 39.8%. The predicted molecular weight of the histidine-tagged CMCase was determined as 56,260 Da. Its Km and Vmax were 9.3 g l-1 and 76.3 g l-1 ·min-1 , respectively. The histidine-tagged CMCase hydrolyzed avicel, carboxymethylcellulose (CMC), filter paper, pullulan, xylan, but there was no detectable activity on cellobiose, p-Nitrophenyl-β-D-glucopyranoside (pNPG)...
January 28, 2019: Preparative Biochemistry & Biotechnology
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