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Biotechnology Letters

William Lorenzi, Fabiany Da Costa Gonçalves, Natália Schneider, Éverton Franco Silva, Fernanda Visioli, Ana Helena Paz, Mauricio Guidi Saueressig
OBJECTIVE: To investigate the effect of adipose tissue-derived mesenchymal stem cell (ASC) administered either systemically or locally in a murine model of bronchiolitis obliterans. RESULTS: When compared to controls, systemic treatment with 10(6) ASCs on D0 and a second dose on D7 significantly prevented tracheal obliteration 28 days after heterotopic tracheal transplantation (median of 94 vs. 16%; P < 0.01). A single dose tended towards less stenosis than controls, but did not reach statistical significance (28 vs...
May 20, 2017: Biotechnology Letters
Jie Yue, Gang Fu, Dawei Zhang, Jianping Wen
OBJECTIVES: To improve heterologous proteins production, we constructed a maltose-inducible expression system in Bacillus subtilis. RESULTS: An expression system based on the promoter for maltose utilization constructed in B. subtilis. Successively, to improve the performance of the P malA -derived system, mutagenesis was employed by gradually shortening the length of P malA promoter and altering the spacing between the predicted MalR binding site and the -35 region...
May 19, 2017: Biotechnology Letters
Hye-Yeong Jo, Sun-Ha Park, Thien-Kim Le, Sang Hoon Ma, Donghak Kim, Taeho Ahn, Young Hee Joung, Chul-Ho Yun
OBJECTIVES: To find the catalytic activities of CYP191A1 from Mycobacterium smegmatis, in which functions of most P450s are unknown, by using a set of reductase systems, peroxides, and various substrates including fatty acids and human drugs. RESULTS: CYP191A1 was functionally expressed in Escherichia coli and purified. Its catalytic activities were examined with fatty acids, chromogenic and fluorogenic substrates, and several human P450 substrates, in the presence of six different types of electron transfer systems, such as rat NADPH-P450 reductase, Candida NADPH-P450 reductase, ferredoxin/ferredoxin reductase, putidaredoxin/putidaredoxin reductase, and peroxides (H2O2 and t-butyl hydroperoxide)...
May 19, 2017: Biotechnology Letters
Lili Wei, Leixi Cao, Yanyan Miao, Shuju Wu, Shumei Xu, Ruisheng Wang, Jun Du, Aihua Liang, Yuejun Fu
OBJECTIVES: To analyze the transcriptome of Spodoptera frugiperda 9 (Sf9) cells infected with AcMNPV or AcMNPV-BmK IT. RESULTS: A comprehensive transcriptome profile for Sf9 cells infected with AcMNPV or AcMNPV-BmK IT is shown. 43127572, 46529744 and 47235310 RNA-Seq profiles permitted the quantification of expression levels for control (C), AcMNPV-BmK IT treatment (ABT) and AcMNPV treatment (AT) groups. There were 997 up-regulated or down-regulated candidate genes with significant different expression level in these treatment groups...
May 19, 2017: Biotechnology Letters
Shu Zhang, Jian Huang, Rong Hu, Guo Guo, Xiaoli Shang, Jianwei Wu
OBJECTIVE: To engineer Pichia pastoris for heterologous production of cellulase from Musca domestica and explore its potential for industrial applications. RESULTS: A new beta-glucosidase gene (bg), encoding 562 amino acids, was cloned from M. domestica by using rapid amplification of cDNA ends. The gene bg was linked to pPICZαA and expressed in P. pastoris with a yield of 500 mg l(-1). The enzyme has the maximum activity with 27.6 U mg(-1) towards cellulose...
May 13, 2017: Biotechnology Letters
Wanchun Han, Congli Zhou, Jiahui Cheng, Mingzhe Pan, Yuejin Hua, Ye Zhao
OBJECTIVES: A 2',3'-cyclic phosphodiesterase gene (drCPDase) has been characterized from Deinococcus radiodurans and is involved in the robust resistance of this organism. RESULTS: Cells lacking 2',3'-cyclic phosphodiesterase gene (drCPDase) showed modest growth defects and displayed increased sensitivities to high doses of various DNA-damaging agents including ionizing radiation, mitomycin C, UV and H2O2. The transcriptional level of drCPDase increased after H2O2 treatment...
May 11, 2017: Biotechnology Letters
Lijun Yang, Litao Zhang, Lihong Yan, Haifeng Zheng, Peifen Lu, Junjun Chen, Jie Dai, Haibiao Sun, Yong Xu, Tao Yang
OBJECTIVE: To assess the stabilities of Arg-Gly-Asp-Trp-Arg (RGDWR, designated as RWR), a new patented antithrombotic small peptide, and its derivative with ω-aminocaprylic acid on its N-terminus (ωRWR). RESULTS: RWR in rat plasma was decreased by between 32 and 48% after 4 h incubation on ice, indicating its instability in plasma. In contrast, ωRWR in plasma remained at 96-107%. Concentration changes were within 6.2% for ωRWR after storage in various conditions...
May 11, 2017: Biotechnology Letters
Ling Bai, Wei He, Tianpeng Li, Cuiting Yang, Yingping Zhuang, Shu Quan
OBJECTIVE: To investigate the application of the TEM-1 β-lactamase protein fragment complementation assay (PCA) in detecting weak and unstable protein-protein interactions as typically observed during chaperone-assisted protein folding in the periplasm of Escherichia coli. RESULTS: The TEM-1 β-lactamase PCA system effectively captured the interactions of three pairs of chaperones and substrates. Moreover, the strength of the interactions can be quantitatively analyzed by comparing different levels of penicillin resistance, and the assay can be performed under 0...
May 10, 2017: Biotechnology Letters
Hoang Thi Minh Hien, Nguyen Cam Ha, Le Thi Thom, Dang Diem Hong
OBJECTIVE: To examine the effect of squalene on liver X receptors (LXRs) that regulate target genes associated with reverse cholesterol transport and thus control whole-body cholesterol homeostasis. RESULTS: To examine the effect of squalene on liver X receptors (LXRs) that regulate target genes associated with reverse cholesterol transport and thus control whole-body cholesterol homeostasis. Squalene significantly stimulated the transactivation of liver X receptor modulator LXRα and LXRβ...
May 10, 2017: Biotechnology Letters
Mona Pourjafar, Massoud Saidijam, Katayoon Etemadi, Rezvan Najafi
OBJECTIVES: To investigate the effect of all-trans retinoic acid (ATRA) on caspase 3 activity, matrix metalloproteinase 2 (MMP-2), and MMP-9 expression and activity as well as in vitro rat bone marrow-derived mesenchymal stem cells (MSCs) migration. RESULTS: The expression of the MMP-2/-9 was at least five times higher in ATRA-treated MSCs (P < 0.001), and MMP-2/-9 activity was enhanced with increasing doses compared to the control MSCs. The caspase three activity was attenuated by ATRA preconditioning...
May 9, 2017: Biotechnology Letters
Aline G Almeida, Rodrigo C V Pinto, C Mark Smales, Leda R Castilho
OBJECTIVES: To develop a recombinant human factor IX (rFIX) formulation equivalent to commercially available products in terms of cake appearance, residual moisture, proportion of soluble aggregates and activity maintenance for 3 months at 4-8 °C. RESULTS: NaCl and low bulking agent/cryoprotectant mass ratio had a negative impact on cake quality upon lyophilisation for a wide range of formulations tested. Particular devised formulations maintained rFIX activity after lyophilization with a similar performance when compared with the rFIX formulated using the excipients reported for a commercially available FIX formulation (Benefix)...
May 8, 2017: Biotechnology Letters
Qianqian Wang, Yixiang Xu, Jiaqi Xu, Xudong Wang, Chen Shen, Yan Zhang, Xiufeng Liu, Boyang Yu, Jian Zhang
OBJECTIVES: To characterize glycosyltransferases from Bacillus subtilis ATCC 6633 and investigate their substrate specificity towards plant polyphenols. RESULTS: Among the cloned and expressed six UDP-glycosyltransferases (BsGT1-6), BsGT-1 showed activity with a wide range of polyphenols: morin, quercetin, alizarin, rehin, curcumin and aloe emodin. The gene of BsGT-1 has an ORF of 1206 bp encoding 402 amino acids. The recombinant enzyme was purified to homogeneity by Ni-NTA affinity chromatograph, and its biochemical characteristics were identified by HPLC-UV/MS, (1)H-NMR and (13)C-NMR...
May 8, 2017: Biotechnology Letters
Evelyn Balsano, Maranda Esterhuizen-Londt, Enamul Hoque, Stephan Pflugmacher Lima
OBJECTIVES: To investigate antioxidative and biotransformation enzyme responses in Mucor hiemalis towards cyanotoxins considering its use in mycoremediation applications. RESULTS: Catalase (CAT), glutathione reductase (GR), and glutathione peroxidase (GPx) in M. hiemalis maintained their activities at all tested microcystin-LR (MC-LR) exposure concentrations. Cytosolic glutathione S-transferase (GST) activity decreased with exposure to 100 µg MC-LR l(-1) while microsomal GST remained constant...
May 8, 2017: Biotechnology Letters
Youzhi Zhou, Ke Liu, Jinsong Zhang, Jianlin Chu, Bingfang He
OBJECTIVE: To improve the stability of β-galactosidase from Bacillus megaterium YZ08 (BMG) in aqueous hydrophilic solvents and promote its application in the galactosylation of natural products. RESULTS: The addition of 5 mM Mg(2+) significantly enhanced the stability of BMG in aqueous hydrophilic solvents, and the half-lives of BMG in these solutions reached 56 min to 208 h, while they were only 7 min to 5.9 h without addition of Mg(2+). Studies on the kinetic parameters in buffer solution and 30% dimethyl sulfoxide (DMSO) indicated that the affinity of BMG to 2-nitrophenyl-β-D-galactopyranoside and its catalytic efficiency (κ cat/K m) increased with the addition of Mg(2+)...
May 3, 2017: Biotechnology Letters
Hanyu Zhu, Dongmei Liu, Yuanyuan Wang, Danfeng Ren, Liesheng Zheng, Liguo Chen, Aimin Ma
OBJECTIVES: To obtain hydrophobin, a Class I hydrophobin gene, Po.hyd from Pleurotus ostreatus, was transformed into the yeast-like cells of Tremella fuciformis using Agrobacterium tumefaciens. RESULTS: The hydrophobin Po.HYD from P. ostreatus was heterogeneously expressed by the yeast-like cells of T. fuciformis. Plasmids harboring the Po.hyd gene driven by endogenous glyceraldehyde-3-phosphate dehydrogenase promoter were transformed by A. tumefaciens. The integration and expression of the rPo...
May 3, 2017: Biotechnology Letters
Li Sun, Quan Yuan, Tianhua Xu, Li Yao, Jiangmin Feng, Jianfei Ma, Lining Wang, Changlong Lv, Danan Wang
OBJECTIVES: To investigate the potential of interleukin (IL)-15 as a novel adjuvant for Mycobacterium tuberculosis (Mtb) antigen 85A (Ag85A) vaccine. RESULTS: C57BL/6 mice were intramuscularly immunized three times with a plasmid expressing the Ag85A-IL-15 fusion protein (pcDNA3.1-Ag85A-IL-15), with the empty pcDNA3.1 vector and the pcDNA3.1-Ag85A as control. Mice vaccinated with pcDNA3.1-Ag85A-IL-15 generated more secretory IgA (sIgA) into their lung (209 ± 21 μg/ml) and acquired an enhanced serum IgG response to Ag85A...
May 3, 2017: Biotechnology Letters
Sandra Luz Martínez-Hernández, Daniel Cervantes-García, Martín Muñoz-Ortega, Liseth R Aldaba-Muruato, Victor M Loera-Muro, Jorge A Ascacio-Martínez, María de Jesús Loera-Arias, Roberto Montes de Oca-Luna, Javier Ventura-Juárez
OBJECTIVE: To generate an immunogenic chimeric protein containing the Entamoeba histolytica LC3 fragment fused to the retrograde delivery domains of exotoxin A of Pseudomonas aeruginosa and KDEL3 for use as an effective vaccine. RESULTS: A codon-optimized synthetic gene encoding the PEΔIII-LC3-KDEL3 fusion construct was designed for expression in Pichia pastoris. This transgene was subcloned into the plasmid pPIC9 for methanol-inducible expression. After transformation and selection of positive-transformed clones by PCR, the expression of the recombinant protein PEΔIII-LC3-KDEL3 was elicited...
May 3, 2017: Biotechnology Letters
I M Gerasymenko, Y V Sheludko
OBJECTIVES: To exploit cold-inducible biochemical processes beneficial for foreign mRNA transcription, translation and storage, as well as protein product stability, during Agrobacterium-mediated transient expression. RESULTS: The efficiency of three different 5'-regulatory sequences to achieve transient expression of the GFP-based reporter gene under chilling conditions (6-8 °C since the 3rd day post inoculation) was compared. We studied the upstream sequences of a cold-inducible Arabidopsis thaliana cor15a gene, the core element of 35S CaMV promoter fused to the TMV omega 5'-UTR, and the synthetic promoter including the 35S core sequence and two binding sites for cold-inducible CBF transcription factors (P_DRE::35S)...
April 24, 2017: Biotechnology Letters
Lili Xu, Xianglong Cheng, Shuangxiu Wu, Quanxi Wang
OBJECTIVES: To improve H2 production, the green algae Chlamydomonas reinhardtii cc849 was co-cultured with Azotobacter chroococcum. RESULTS: The maximum H2 production of the co-culture was 350% greater than that of the pure algal cultures under optimal H2 production conditions. The maximum growth and the respiratory rate of the co-cultures were about 320 and 300% of the controls, and the dissolved O2 of co-cultures was decreased 74%. Furthermore, the in vitro maximum hydrogenase activity of the co-culture was 250% greater than that of the control, and the in vivo maximum hydrogenase activity of the co-culture was 1...
April 21, 2017: Biotechnology Letters
Qiujin Liang, Zhengzhi Huang, Yuan Zhang, Hongtao Li
OBJECTIVE: To fabricate an active and stable enzyme through covalent immobilization, a Ubl-specific protease (Ulp1) was used to cleave small ubiquitin-like modifier (SUMO) fusion proteins. RESULTS: We immobilized Ulp1 on N-hydroxysuccinimide (NHS)-activated Sepharose with a coupling efficiency of 1.7 mg/ml. The immobilized Ulp1 maintains 95% substrate-cleavage ability and significantly enhances pH and thermal stability, especially can withstand pH of 10.5. Besides resistance against some small molecules, the immobilized Ulp1 can tolerate 15% (v/v) DMSO and 20% (v/v) ethanol...
April 21, 2017: Biotechnology Letters
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