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Biotechnology Letters

Jianghao Chen, Wei Liu, Jinxiu Zhao, Cong Sun, Jie Chen, Kaijin Hu, Linlin Zhang, Yuxiang Ding
OBJECTIVES: To investigate the therapeutic effect of gelatin microspheres containing different concentrations of calcitonin gene-related peptide (CGRP) or substance P on repairing bone defects in a rabbit osteoporosis model. RESULTS: Gelatin microspheres containing different concentrations of CGRP or substance P promoted osteogenesis after 3 months in a rabbit osteoporotic bone defective model. From micro-computed tomography imaging results, 10 nM CGRP was optimal for increasing the trabecular number and decreasing the trabecular bone separation degree; similar effects were observed with the microspheres containing 1 µM substance P...
December 1, 2016: Biotechnology Letters
Yingtong Zhang, Haiqin Chen, Eusebio Navarro, Sergio López-García, Yong Q Chen, Hao Zhang, Wei Chen, Victoriano Garre
OBJECTIVES: To generate lycopene-overproducing strains of the fungus Mucor circinelloides with interest for industrial production and to gain insight into the catalytic mechanism of lycopene cyclase and regulatory process during lycopene overaccumulation. RESULTS: Three lycopene-overproducing mutants were generated by classic mutagenesis techniques from a β-carotene-overproducing strain. They carried distinct mutations in the carRP gene encoding lycopene cyclase that produced loss of enzymatic activity to different extents...
December 1, 2016: Biotechnology Letters
Nicole Werner, Miriam Dreyer, Wenke Wagner, Nicolas Papon, Steffen Rupp, Susanne Zibek
OBJECTIVES: To explore Candida guilliermondii for the production of long-chain dicarboxylic acids (DCA), we performed metabolic pathway engineering aiming to prevent DCA consumption during β-oxidation, but also to increase its production via the ω-oxidation pathway. RESULTS: We identified the major β- and ω-oxidation pathway genes in C. guilliermondii and performed first steps in the strain improvement. A double pox disruption mutant was created that slowed growth with oleic acid but showed accelerated DCA degradation...
November 30, 2016: Biotechnology Letters
Chih-Yueh Liu, Chang-Ching Weng, Chih-Hsiang Lin, Chiou-Ying Yang, Kwok-Kong Tony Mong, Yaw-Kuen Li
OBJECTIVES: A Neissaria bacterial pilus sugar, bacillosamine, was synthesized and, for the first time, used as a probe to screen a single-chain variable fragment (scFv). RESULTS: Four Neisseria, Neisseria gonorrhoeae, Neisseria meningitidis, Neisseria sicca and Neisseria subflava, and two negative controls, Streptococcus pneumoniae and Escherichia coli, were tested through ELISA, immunostaining and gold nanoparticle immunological assay. All results indicated that the selected scFv is feasible for the specific detection of Neisseria species via the recognition of bacillosamine...
November 25, 2016: Biotechnology Letters
Hee-Jung Lim, Thi Thanh Hanh Nguyen, Nahyun M Kim, Jun-Seong Park, Tae-Su Jang, Doman Kim
OBJECTIVES: To determine the inhibitory activities of flavonoids against NS2B-NS3 protease of ZIKA virus (ZIKV NS2B-NS3(pro)) expressed in Escherichia coli BL21 (DE3) and their structure activity relationship. RESULTS: ZIKV NS2B-NS3(pro) was expressed in E. coli BL21(DE3) as a 35 kDa protein. It had a K m of 26 µM with the fluorogenic peptide Dabcyl-KTSAVLQSGFRKME-Edan. The purified ZIKV NS2B-NS3(pro) was used for inhibition and kinetic assays to determine the activities of 22 polyphenol compounds...
November 24, 2016: Biotechnology Letters
Eugenio Spadoni Andreani, Federica Villa, Francesca Cappitelli, Anna Krasowska, Piotr Biniarz, Marcin Łukaszewicz, Francesco Secundo
OBJECTIVES: To investigate the ability of the proteases, subtilisin and α-chymotrypsin (aCT), to inhibit the adhesion of Candida albicans biofilm to a polypropylene surface. RESULTS: The proteases were immobilized on plasma-treated polypropylene by covalently linking them with either glutaraldehyde (GA) or N'-diisopropylcarbodiimide (DIC) and N-hydroxysuccinimide (NHS). The immobilization did not negatively affect the enzyme activity and in the case of subtilisin, the activity was up to 640% higher than that of the free enzyme when using N-acetyl phenylalanine ethyl ester as the substrate...
November 23, 2016: Biotechnology Letters
Jie Wang, Bhaskar Roy
OBJECTIVE: To characterize transcriptome-wide lincRNAs of Hela-S3 cell line by analyzing RNA sequencing data to provide a foundation for further functional verification and clinical application of cervical carcinoma development. RESULTS: Single-cell RNA sequencing data of 37 Hela-S3 cells were analysed. On average, 511 lincRNAs were expressed in each cell. Comparing the expression difference of the lincRNAs and protein-coding genes, we found that lincRNAs expression displayed more cell specificity than that of protein-coding genes (t-test, P<2...
November 23, 2016: Biotechnology Letters
Chelsea Ledford, Monica McMahon, Ashley Whitesell, Ghalib Khan, Suneel K Kandagatla, Dow P Hurst, Patricia H Reggio, Gregory M Raner
OBJECTIVE: To develop a model for binding and catalysis associated with the stimulation of 4-fluorophenol (4-FP) oxidation in the presence of long chain aldehydes by the enzymatic catalyst, cytochrome P450BM3-F87G. RESULTS: A variation of the Michaeli-Menten kinetic model was employed to describe interactions at the active site of the enzyme, along with computer aided modeling approaches. In addition to the hydroquinone product arising from de-fluorination of 4-FP, a second product (p-fluorocatechol) was also observed and, like the hydroquinone, its rate of formation increased in the presence of the aldehyde...
November 18, 2016: Biotechnology Letters
Yu-Chen Zhang, Long Jin, Hai-Ying Zhu, Qing Guo, Xiao-Chen Li, Guang-Lei Zhang, Xiao-Xu Xing, Mei-Fu Xuan, Qi-Rong Luo, Zhao-Bo Luo, Jun-Xia Wang, Cheng-Du Cui, Wen-Xue Li, Zheng-Yun Cui, Xi-Jun Yin, Jin-Dan Kang
OBJECTIVE: The aim of this study was to investigate the developmental competence of oocytes parthenogenetically activated by an electric pulse (EP) and treated with anisomycin and to determine whether this method is applicable to somatic cell nuclear transfer (SCNT). RESULTS: Embryos derived from porcine oocytes parthenogenetically activated by an EP and treatment with 0.01 µg/mL anisomycin had a significantly improved in vitro developmental capacity. Furthermore, 66...
November 18, 2016: Biotechnology Letters
Yu Nie, Peiqing Cong, Xiaofeng Liu, Min Wang, Yaosheng Chen, Zuyong He
OBJECTIVES: To develop an in vitro method for rapid evaluation of the capability of a designed single guide RNAs (sgRNAs) to guide Cas9 nucleases to cleave target loci in mammalian cells. RESULTS: We constructed a Cas9/sgRNA plasmid with two SP6 promoters to simultaneously express Cas9 nuclease and the sgRNA and a negative selection plasmid harbouring a target site of the sgRNA. After co-transforming chemically competent E. coli DH5α cells with the two plasmids, the transformants were plated at a low density on two LB plates: one containing only ampicillin and the other containing both ampicillin and chloramphenicol...
November 17, 2016: Biotechnology Letters
Yasodha Manandhar, Wei Wang, Jin Inoue, Nobuhiro Hayashi, Takanori Uzawa, Yutaka Ito, Toshiro Aigaki, Yoshihiro Ito
OBJECTIVES: We examined the importance of aptamer usage under the same condition as the selection process by employing the previously selected aptamers for calmodulin (CaM) which includes a non-natural fluorogenic amino acid, 7-nitro-2,1,3-benzoxadiazole. RESULTS: We added five amino acids at the N-terminus which was employed for the selection and then we tested the affinity and selectivity for CaM binding. Surface plasmon resonance and fluorescence measurements showed that the additional amino acids for one of the aptamers drastically improved binding affinity to CaM, indicating the importance of aptamer use under the same conditions as the selection process...
November 17, 2016: Biotechnology Letters
Gábor Márk Tardy, Bálint Lóránt, Máté Lóka
OBJECTIVES: Power production characteristics and substrate concentration dependence of voltage have been investigated together with the determination of kinetic constants in two-chambered mediator-less microbial fuel cells (MFC) for acetate and peptone substrates. RESULTS: At 500 mg DOC l(-1) (dissolved organic carbon), power densities normalized to the anode surface of 112 mW m(-2) with acetate and 114 mW m(-2) with peptone as electron donor were attained by applying cathodes with a Pt catalyst layer...
November 17, 2016: Biotechnology Letters
Shaoqing Yang, Qiaojuan Yan, Qingdan Bao, Jingjing Liu, Zhengqiang Jiang
OBJECTIVES: To identify novel pullulanases from microorganisms and to investigate their biochemical characterizations. RESULTS: A novel pullulanase gene (BmPul) from Bacillus megaterium WW1210 was cloned and heterologously expressed in Escherichia coli. The gene has an ORF of 2814 bp encoding 937 amino acids. The recombinant pullulanase (BmPul) was purified to homogeneity and biochemically characterized. BmPul has an MW of approx. 112 kDa as indicated by SDS-PAGE...
November 17, 2016: Biotechnology Letters
Shenghai Wang, Mengjie Duan, Yalan Liu, Sen Fan, Xiaoshan Lin, Yi Zhang
OBJECTIVE: To breed Aspergillus oryzae strains with high fructosyltransferase (FTase) activity using intraspecific protoplast fusion via genome-shuffling. RESULTS: A candidate library was developed using UV/LiCl of the conidia of A. oryzae SBB201. By screening for enzyme activity and cell biomass, two mutants (UV-11 and UV-76) were chosen for protoplast fusion and subsequent genome shuffling. After three rounds of genome recombination, a fusion mutant RIII-7 was obtained...
November 16, 2016: Biotechnology Letters
Hai Zhang, Fang Wang, Yahua Hu
OBJECTIVES: To study the roles of STARD13 in cellular apoptosis of hepatocellular carcinoma (HCC). RESULTS: Quantitative real-time PCR and immunohistochemistry analyses showed that the expression levels of STARD13 and Fas were lower in clinical HCC tissues than in normal tissues and were positively correlated, which is consistent with the results analyzed by The Cancer Genome Atlas (TCGA) data. Patients with higher STARD13 or Fas expression levels had longer overall survival...
November 14, 2016: Biotechnology Letters
Yuzhen Wei, Jianjing Sun, Xingang Li
OBJECTIVE: To elucidate the molecular mechanism of microRNA-215 (miR-215) in the migration and invasion of high grade glioma. RESULTS: 42 Patients were analysed for clinicopathological characteristics. qRT-PCR showed that miR-215 was up-regulated in glioma tissues compared with non-neoplastic brain tissues (P < 0.05). The up-regulated miR-215 was closely associated with high grade glioma (P < 0.01) and poor overall survival (P < 0.01). Transwell assay showed that re-expression of miR-215 enhanced migration and invasion of glioma cells...
November 11, 2016: Biotechnology Letters
Jorge Xool-Tamayo, Germán Serrano-Gamboa, Miriam Monforte-González, Gumersindo Mirón-López, Felipe Vázquez-Flota
OBJECTIVE: To analyze berberine and sanguinarine biosynthetic capacities of both in vitro shoot and root cultures of Argemone mexicana and tissues from entire plants at different developmental stages. RESULTS: Berberine and sanguinarine were equally distributed in roots and aerial tissues of developing plantlet whereas, in juvenile plants, sanguinarine was only detected in roots. This alkaloid distribution was consistent with that of biosynthetic transcripts in juvenile plants...
November 11, 2016: Biotechnology Letters
Haizhou Long, Yi Yao, Shouhong Jin, Yingting Yu, Xiongbing Hu, Fengfeng Zhuang, Hanshuo Zhang, Qiong Wu
OBJECTIVE: RNAe is a new method that enhances protein expression at the post-transcriptional level. RNAe utility was further explored to improve endogenous protein expression. RESULTS: Transgenic mice were created by targeting RNAe to growth hormone gene into the C57/BL mouse genome by transposon mediated integration; the mice showed a heavier body weight and longer body length compared with normal mice. RNAe can also be used for gene therapy through the delivery of in vitro transcribed RNA...
November 4, 2016: Biotechnology Letters
Godwin E Oyiwona, James Ogbonna, Chukwudi Uzoma Anyanwu, So Ishizaki, Zen-Ichiro Kimura, Satoshi Okabe
OBJECTIVE: To investigate a syntrophic interaction between Geobacter sulfurreducens and hydrogenotrophic methanogens in sludge-inoculated microbial fuel cell (MFC) systems running on glucose with an improved electron recovery at the anode. RESULTS: The presence of archaea in MFC reduces Coulombic efficiency (CE) due to their electron scavenging capability but, here, we demonstrate that a syntrophic interaction can occur between G. sulfurreducens and hydrogenotrophic methanogens via interspecies H2 transfer with improvement in CE and power density...
November 3, 2016: Biotechnology Letters
Karin Bjerre, Mette D Cantor, Jan V Nørgaard, Hanne D Poulsen, Karoline Blaabjerg, Nuria Canibe, Bent B Jensen, Birgitte Stuer-Lauridsen, Bea Nielsen, Patrick M F Derkx
OBJECTIVES: To generate tryptophan-overproducing Bacillus subtilis strains for in situ use in pigs, to reduce the feed cost for farmers and nitrogen pollution. RESULTS: A novel concept has been investigated-to generate B. subtilis strains able to produce tryptophan (Trp) in situ in pigs. Mutagenesis by UV was combined with selection on Trp and purine analogues in an iterative process. Two mutants from different wild types were obtained, mutant 1 (M1) produced 1 mg Trp/l and mutant 2 (M2) 14 mg Trp/l...
November 3, 2016: Biotechnology Letters
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