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Preparative Biochemistry & Biotechnology

Amanda Reges de Sena, Flávio Manoel Barros Oliveira, Tonny Cley Campos Leite, Talita Camila Evaristo da Silva Nascimento, Keila Aparecida Moreira, Sandra Aparecida de Assis
The aims of the current study are to assess the influence of polyethylene glycol (PEG) concentration, molar mass, pH and citrate concentrations on aqueous biphasic systems based on 2(4) factorial designs, as well as to check their capacity to purify tannase secreted by Aspergillus tamarii URM 7115. Tannase was produced through submerged fermentation at 26 °C for 67 h in Czapeck-Dox modified broth and added with yeast extract and tannic acid. The factorial design was followed to assess the influence of PEG molar mass (MPEG 600; 4000 and 8000 g/ mol), and PEG (CPEG 20...
August 31, 2017: Preparative Biochemistry & Biotechnology
Juan Daniel Rivaldi, Ana Karine F Carvalho, Leyvison Rafael V da Conceição, Heizir F de Castro
Increased costs and limited availability of traditional lipid sources for biodiesel production encourages researchers to find more sustainable feedstock at low prices. Microbial lipid stands out as feedstock replacement for vegetable oil to convert fatty acid esters (FAEs). In this study, the potential of three isolates of filamentous fungi (Mucor circinelloides URM 4140, M. hiemalis URM 4144 and Penicillium citrinum URM 4126) has been assessed as single-cell oil (SCO) producers. M. circinelloides 4140 had the highest biomass concentration with lipid accumulation of up to 28 wt% at 120 h of cultivation...
August 31, 2017: Preparative Biochemistry & Biotechnology
Thomas Langer, Carsten Corvey, Katja Kroll, Oliver Boscheinen, Thomas Wendrich, Werner Dittrich
Glycosylation is one of the most complex posttranslational modifications and may have significant influence on the proper function of the corresponding proteins. Bacteria and yeast are, because of easy handling and cost reasons, the most frequently used systems for recombinant protein expression. Bacteria generally do not glycosylate proteins and yeast might tend to hyperglycosylate. Insect cell and mammalian cell based expression system are able to produce complex N-glycosylation structures, but are more complex to handle and more expensive...
August 31, 2017: Preparative Biochemistry & Biotechnology
Ali Osman Adıgüzel, Münir Tunçer
A total of approximately 400 bacterial strains were isolated from 73 plastic wastes collected from 14 different regions. Nineteen isolates, that form clear zones both on Tributyrin and PCL Agar, were identified based on 16S rRNA gene sequences. Among these, Bacillus sp. KY0701 that caused highest weight loss of PCL films in MSM, was selected for cutinase production. The highest enzyme activity (15 U/mL) was obtained after 4 days of incubation at 50 °C, pH 7.0 and 200 rpm in a liquid medium containing 1.5% (w/v) apple cutin and 0...
August 31, 2017: Preparative Biochemistry & Biotechnology
Mansoureh Fazelahi, Vahid Kia, Hooman Kaghazian, Mahdi Paryan
One of the most important biopharmaceuticals worldwide is recombinant streptokinase, which is usually produced in Escherichia coli (E.coli). During the biopharmaceutical purification steps, residual host cell DNA may remain in the product, and since residual host cell DNA is considered as a contamination and risk factor, it is necessary to purify the recombinant product as much as possible. Therefore, it is necessary to control the production procedure in order to exclude any possible contamination. The aim of the present study was to develop a highly specific and sensitive quantitative real-time PCR-based method to determine the amount of E...
August 31, 2017: Preparative Biochemistry & Biotechnology
Hooi Xian Lee, Fisal Ahmad, Bahruddin Saad, Mohd Nazri Ismail
Date fruits are well known to be very nutritious. Nevertheless, the protein content of the fruit, particularly the seed and flesh is still understudied, largely due to their difficult physical characteristics. This study was conducted to compare three different protein extraction methods which were the TCA-acetone (TCA-A), phenol (Phe), and TCA-acetone-phenol (TCA-A-Phe), and to perform proteomic analysis on date palm seed and flesh. Phe extraction method showed the highest protein yields for both seed (8.26 mg/ g) and flesh (1...
August 31, 2017: Preparative Biochemistry & Biotechnology
Mu-Tan Luo, Chao Huang, Xue-Fang Chen, Qian-Lin Huang, Gao-Xiang Qi, Lan-Lan Tian, Lian Xiong, Hai-Long Li, Xin-De Chen
Biomass acid hydrolysate of oleaginous yeast Trichosporon cutaneum after microbial oil extraction was applied as substrate for bacterial cellulose (BC) production by Komagataeibacter xylinus (also named as Gluconacetobacter xylinus previously) for the first time. BC was synthesized in static culture for ten days and the maximum BC yield (2.9 g/ L) was got at the 4th day of fermentation. Most carbon sources in the substrate (glucose, mannose, formic acid, acetic acid, etc.) can be utilized by K. xylinus...
August 31, 2017: Preparative Biochemistry & Biotechnology
Ngangoum Eric Serge, Mezajoug Kenfack Laurette Blandine, Sanjit Kumar, Tchiégang Clergé, Mookambeswaran Vijayalakshmi
A protease from fresh leaves of Abrus precatorius was purified using two classical chromatography techniques: ion exchange (DEAE-Sepharose) and Gel filtration (Sephadex G-75). The Purified protease showed a molecular weight of ∼ 28 kDa on sodium dodecyl gel electrophoresis (SDS-PAGE).The optimum pH and temperature for the purified protease was 8 and 40°C respectively. The purified protease was stable throughout a wide temperature range from 10°C to 80°C and pH from 2 to 12. Protease activity was inhibited in presence of Co(2+), Ni(2+), Hg(2+) and Zn(2+) while its activity has increased in presence of Ca(2+) and Mg(2+)...
August 31, 2017: Preparative Biochemistry & Biotechnology
Hui Li, Jiao Xue, Wanlong Zhou, Yajie Sun, Wei Liu, Weiping Lin, Ao Liu, Aihuan Song, Hu Zhu
As one kind of important secondary metabolites produced by Inonotus baumii, flavones can be applied in food, medicine and other industries due to their biological activities such as anti-oxidant, anti-cancer, and anti-baceterial activity. In order to enhance total flavone production in submerged fermentation of I. baumii, three different strategies, optimization of fermentation parameters by statistical designs including Plackett-Burman design and Response Surface Methodology, addition of precursors and elicitors and two-phase culture were used...
August 31, 2017: Preparative Biochemistry & Biotechnology
Aila Riany de Brito, Nadabe Dos Santos Reis, Tatielle Pereira Silva, Renata Cristina Ferreira Bonomo, Ana Paula Trovatti Uetanabaro, Sandra Aparecida de Assis, Erik Galvão Paranhos da Silva, Elizama Aguiar-Oliveira, Julieta Rangel Oliveira, Marcelo Franco
Endoglucanase production by Aspergillus oryzae ATCC 10124 cultivated in rice husks or peanut shells was optimized by experimental design as a function of humidity, time and temperature. The optimum temperature for the endoglucanase activity was estimated by a univariate analysis (one factor at the time) as 50 °C (rice husks) and 60 °C (peanut shells), however, by a multivariate analysis (synergism of factors) it was determined a different temperature (56 °C) for endoglucanase from peanut shells. For the optimum pH, values determined by univariate and multivariate analysis were: 5...
August 31, 2017: Preparative Biochemistry & Biotechnology
Mohammad Hossein Hedayati, Dariush Norouzian, Mahdi Aminian, Shahram Teimourian, Reza Ahangari Cohan, M Reza Khorramizadeh
BACKGROUND: Oxidative degradation of human recombinant erythropoietin (hrEPO) may occur in manufacturing process or therapeutic applications. This unfavorable alteration may render EPO inefficient or inactive. We investigated the effect of Methionine/54 oxidative changes on the amino acid sequences, glycoform distribution an d biological activity of hrEPO. METHODS: Mass spectrometry was applied to verify the sequence and determine the methionine oxidation level of hrEPO...
August 21, 2017: Preparative Biochemistry & Biotechnology
Yanlu Zhang, Liehua Wu, Jingai Yu, JianFeng Mei, Yu Yi, JianShu Chen, Guoqing Ying
Human thymic stromal lymphopoietin (hTSLP) protein plays a central role in inflammation. Characterizing properties of hTSLP requires a recombinant overexpression system that produces correctly folded, active hTSLP. In this report, an efficient overexpression system for production of hTSLP was developed. We constructed expression plasmids of the full-length hTslp gene with or without the signal peptide and transformed the plasmids into E. coli. The design of the recombinant proteins included an N-terminal His-tag, which facilitated purification...
August 17, 2017: Preparative Biochemistry & Biotechnology
Seenivasan Ayothiraman, Sathyanarayana N Gummadi, Tapobrata Panda
The elution characteristics of lovastatin were studied on varying the composition of mobile phase in both isocratic and gradient elution modes to comprehend the role of organic modifier and acidifier on the overall analysis time and retention time of individual forms of lovastatin. Acetonitrile has influenced on the overall analysis time, whereas the acidifier determines the retention time of hydroxy acid form of lovastatin and the retention time gap between the individual forms. A combination of acetonitrile and 0...
August 17, 2017: Preparative Biochemistry & Biotechnology
Yunier Rodríguez-Álvarez, Klaudia Martínez-Cordovez, Alexey Llopiz-Arzuaga, Yassel Ramos-Pérez, Vladimir Besada-Pérez, Dayana García-Lines, Alicia Santos-Savio
Recombinant simian IL-15 (siIL-15) was obtained for the preclinical assessment of an anti-human IL-15 vaccine. For this purpose, the cDNA from peripheral blood mononuclear cells of a Macaca fascicularis monkey was cloned into a pIL-2 vector. The siIL-15 was expressed in E. coli strain W3110 as an insoluble protein which accounted for 13% of the total cellular proteins. Inclusion bodies were solubilized in an 8 M urea solution, which was purified by ion exchange and reverse phase chromatography up to 92% purity...
August 17, 2017: Preparative Biochemistry & Biotechnology
Jingping Ge, Changli Wang, Shoufeng Huang, Renpeng Du, Kun Liu, Gang Song, Wenxiang Ping
Natamycin has been widely applied in medical treatments and food protection widely due to its effective inhibition to the growth of yeast and mold. As polyene macrolide antibiotic, the biosynthesis pathway of natamycin is relatively clear. To regulate the biosynthesis of natamycin, additions of precursors affecting cell growth and natamycin production were investigated. The results showed that 0.003% (w/v) potassium ferrocyanide and sodium propionate: n-butanol at a ratio of 4:1 was added into the broth at 0 h and 24 h respectively and they contributed to yield natamycin, reaching 1...
August 17, 2017: Preparative Biochemistry & Biotechnology
Ayşe Ezgi Ünlü, Brinda Prasad, Kishan Anavekar, Paul Bubenheim, Andreas Liese
Flavonoids are polyphenolic secondary plant metabolites which possess antioxidant and anti-inflammatory properties. Besides, they have been shown to exhibit increased antioxidant properties in their polymerized form. Catechins are one of the attractive class of flavonoids which belong to the group of flavan-3-ols. Polymerization of catechins have been investigated in numerous studies indicating the requirement of certain amount of organic solvent to provide the solubility of the monomer. However, many research projects have been conducted recently in order to replace toxic organic contaminants of the processes with environmentally friendly solvents...
August 17, 2017: Preparative Biochemistry & Biotechnology
Katarzyna Struszczyk-Świta, Łukasz Stańczyk, Mirosława Szczęsna-Antczak, Tadeusz Antczak
Mucor circinelloides IBT-83 mycelium that exhibits both lipolytic (AL) and chitosanolytic (ACH) activities was immobilized into polyurethane foam (PUF) in a 30 L laboratory fermenter. The process of immobilization was investigated in terms of: the carrier porosity, its type, amount and shape, location inside the fermenter, mixing and aeration parameters during the culture, as well as downstream processing operations. The selected conditions allowed for immobilization of approx. 7 g of defatted and dried mycelium in 1 g of carrier, i...
August 17, 2017: Preparative Biochemistry & Biotechnology
Guoqiang Chen, Jiming Wang, Yulong Su, Youshuang Zhu, Ge Zhang, Hongwei Zhao, Haobao Liu, Ying Yang, Rui Nian, Haibo Zhang, Yuxi Wei, Mo Xian
Pullulan with different molecular-weight could be applied in various fields. A UV-induced mutagenesis Aureobasidium pullulans UVMU6-1 was obtained from the strain A. pullulans CGMCC3.933 for the production of low-molecular-weight pullulan. Firstly, the obtained polysaccharide from A. pullulans UVMU6-1 was purified and identified to be pullulan with TLC, FT-IR and NMR. Then, culture medium and conditions for this strain were optimized by flask fermentation. Based on the optimized medium and culture conditions (pH 4, addition of 4 g/L Tween 80 for 96 h of cultivation), continuously fermentation was performed...
July 18, 2017: Preparative Biochemistry & Biotechnology
Benlin Qin, Xuecong Liu, Haiming Cui, Yue Ma, Zimin Wang, Jing Han
In this study, an efficient ultrasound-assisted aqueous two-phase extraction (UA-ATPE) method was employed for the extraction of anthocyanins from Lycium ruthenicum Murr(LRMAs). An ethanol/ammonium sulfate system was chosen for the aqueous two-phase system (ATPS) due to its fine partitioning and recycling behaviors. Single-factor experiments were conducted to determine the optimized composition of the system, and the response surface methodology (RSM) was used for the further optimization of the ultrasound-assisted aqueous two-phase extraction...
July 14, 2017: Preparative Biochemistry & Biotechnology
Ping Yu, Pengzhi Zhu
S-adenosyl-L-methionine (SAM) has important applications in many fields including the chemical therapy and the pharmaceutical industry. In this study, the recombinant Escherichia coli strain was constructed for effective production of SAM by introducing the SAM synthase gene (metK). This strain produced 34.5 mg/l of SAM in basic medium in shake flask. Yeast extract, pH and loaded volume had a significant positive effect on the yield of SAM. Their optimal values were 35 g/l, 7.5 and 30 ml, respectively...
July 14, 2017: Preparative Biochemistry & Biotechnology
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