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Biotechnology Letters

Vahid Jalali Javaran, Alireza Shafeinia, Mokhtar Jalali Javaran, Esmaeil Ghasemi Gojani, Malihe Mirzaee
OBJECTIVE: To use a transient expression system to express a truncated human tissue plasminogen activator (K2S) gene in cucurbit plants. RESULTS: The recombinant tissue plasminogen activator protein (K2S form) was expressed in active form in cucurbit plants. Its molecular weight was 43 kDa. The plant-derived rt-PA was determined using goat anti-rabbit antibody by western blotting. Among the infected lines, the highest expression of rt-PA was 62 ng/100 mg per leaf tissue as measured by ELISA...
January 16, 2017: Biotechnology Letters
Hyo Sang Jo, Eun Ji Yeo, Min Jea Shin, Yeon Joo Choi, Hyeon Ji Yeo, Su Bin Cho, Jung Hwan Park, Chi Hern Lee, Won Sik Eum, Soo Young Choi
OBJECTIVES: To identify the protective effect of DJ-1 protein against oxidative stress-induced HepG2 cell death, we used cell-permeable wild type (WT) and a mutant (C106A Tat-DJ-1) protein. RESULTS: By using western blotting and fluorescence microscopy, we observed WT and C106A Tat-DJ-1 proteins were efficiently transduced into HepG2 cells. Transduced WT Tat-DJ-1 proteins increased cell survival and protected against DNA fragmentation and intracellular ROS generation levels in H2O2-exposed HepG2 cells...
January 10, 2017: Biotechnology Letters
Xianzhang Jiang, Hongjiao Liu, Yongchao Niu, Feng Qi, Mingliang Zhang, Jianzhong Huang
No abstract text is available yet for this article.
January 10, 2017: Biotechnology Letters
Chenghong Wang, Jiguo Qiu, Youjian Yang, Jinwei Zheng, Jian He, Shunpeng Li
OBJECTIVE: To identify and characterize a novel aryloxyphenoxypropionate (AOPP) herbicide-hydrolyzing carboxylesterase from Aquamicrobium sp. FPB-1. RESULTS: A carboxylesterase gene, fpbH, was cloned from Aquamicrobium sp. FPB-1. The gene is 798 bp long and encodes a protein of 265 amino acids. FpbH is smaller than previously reported AOPP herbicide-hydrolyzing carboxylesterases and shares only 21-35% sequence identity with them. FpbH was expressed in Escherichia coli BL21(DE3) and the product was purified by Ni-NTA affinity chromatography...
January 5, 2017: Biotechnology Letters
Joon Serk Seo, Byung Sub Min, Yeon Jung Kim, Jong Moon Cho, Gi-Seong Kwon, Byeong-Pil Lim, Shin-Jae Chang, Dong-Il Kim
OBJECTIVES: To evaluate the characteristics of a novel human cell line, F2N78, including growth performance, physicochemical properties, and biological activity via direct comparison with CHO cells. RESULTS: The culture performance and physicochemical properties of antibodies produced from F2N78 and CHO cells were compared. For charge variants, antibodies produced from F2N78 cells contained a greater acidic charge variants than CHO cells. Regarding main glycoforms, degree of galactosylation was 52% in CT-A produced from F2N78 cells compared to CHO cells (37%)...
January 4, 2017: Biotechnology Letters
Lu-Lu Hou, Feng Liu, Xiaonan Zang, Xuecheng Zhang, Bangxiang He, Yating Ding, Xinwei Song, Dongfang Xiao, Haitao Wang
OBJECTIVES: To optimize the cultivation media for the growth rate of Haematococcus pluvialis and to study the transcription regulation of the algal nitrate reductase (NR), a key enzyme for nitrogen metabolism. RESULTS: The NR gene from H. pluvialis hd7 consists of 5636 nucleotides, including 14 introns. The cDNA ORF is 2718 bp, encoding a 905 aa protein with three conserved domains. The NR amino acids of H. pluvialis hd7 are hydrophilic and have similarity of 72% compared to that of Dunaliella...
January 4, 2017: Biotechnology Letters
Chunlong Sun, Hongtao Zhang, Wen Du, Baoqin Wang, Min Ji
OBJECTIVE: Methionine is a valid target for the treatment of cancer and to achieve in vivo imaging and early diagnosis of tumors, we have synthesized near-infrared (NIR) fluorochrome IR-822-labeled methionine (IR-822-Met). RESULTS: NIR fluorescent dye IR-822 was conjugated with methionine through its amide bond. It had low toxicity to normal cell/tissues. In vitro and in vivo studies demonstrated its high targeting capability to tumors. The results support the potential of using ligand-modified methionine probe for tumor diagnosis and targeted therapy...
January 3, 2017: Biotechnology Letters
Amélie Dukunde, Dominik Schneider, Mingji Lu, Silja Brady, Rolf Daniel
OBJECTIVES: To investigate the properties of a novel metagenome-derived member of the hormone-sensitive lipase family of lipolytic enzymes. RESULTS: A forest soil metagenome-derived gene encoding an esterase (Est06) belonging to the hormone-sensitive lipase family of lipolytic enzymes was subcloned, heterologously expressed and characterized. Est06 is a polypeptide of 295 amino acids with a molecular mass of 31 kDa. The deduced protein sequence shares 61% similarity with a hypothetical protein from the marine symbiont Candidatus Entotheonella sp...
January 2, 2017: Biotechnology Letters
Ongey Elvis Legala, Hüseyin Yassi, Stephan Pflugmacher, Peter Neubauer
The intrinsic qualities of lanthipeptides for their use as therapeutic drugs present several challenges because of their properties, which include stability, solubility and bioavailability, which, under physiological conditions, are very low. Researches have encouraged clinical evaluation of a few compounds, such as mutacin 1140, microbisporicin, actagardine and duramycin, with pharmacokinetic profiles showing rapid distribution and elimination rates, good bioavailability and fecal excretion, as well as high protein binding...
January 2, 2017: Biotechnology Letters
Feng Cheng, Jian-Miao Xu, Chao Xiang, Zhi-Qiang Liu, Li-Qing Zhao, Yu-Guo Zheng
OBJECTIVE: To develop a practically simple and robust multi-site saturation mutagenesis (MSSM) method that enables simultaneously recombination of amino acid positions for focused mutant library generation. RESULTS: A general restriction enzyme-free and ligase-free MSSM method (Simple-MSSM) based on prolonged overlap extension PCR (POE-PCR) and Simple Cloning techniques. As a proof of principle of Simple-MSSM, the gene of eGFP (enhanced green fluorescent protein) was used as a template gene for simultaneous mutagenesis of five codons...
January 2, 2017: Biotechnology Letters
Haiyan Sun, Ming Peng
OBJECTIVES: To investigate the role of maltose stearic acid ester (MSAE) for improving raw-starch-digesting glucoamylase (RSDG) production by Aspergillus niger F-01 and its regulatory mechanism. RESULTS: RSDG activity was enhanced 8.3-fold by adding 5 g MSAE to 1 l basal medium at the beginning of the culture. RSDG biosynthesis nearly ceased by adding 100 μg actinomycin D/ml or 50 μg cycloheximide/ml in a 48 h culture in the basal medium. RSDG biosynthesis was restarted by adding MSAE in the culture repressed by actinomycin D...
January 2, 2017: Biotechnology Letters
Changzheng Wang, Chuanhu Qiao
OBJECTIVES: To determine the role of miR-190b in radio-sensitivity of gastric cancer (GC). RESULTS: In radio-resistant GC cells, down-regulation of miR-190b and up-regulation of Bcl-2 were observed. The protein expression of Bcl-2 was negatively regulated by miR-190b. Overexpression of miR-190b significantly decreased cell viability and enhanced radio-sensitivity of GC cells. Of note, these effects of miR-190b on GC cells radio-sensitivity were abolished by Bcl-2...
January 2, 2017: Biotechnology Letters
Liang Kang, Cao Yang, Huipeng Yin, Kangcheng Zhao, Wei Liu, Wenbin Hua, Kun Wang, Yu Song, Ji Tu, Shuai Li, Rongjin Luo, Yukun Zhang
OBJECTIVES: To determine the role of microRNA-15b (miR-15b) in interleukin-1 beta (IL-1β)-induced extracellular matrix (ECM) degradation in the nucleus pulposus (NP). RESULTS: MiR-15b was up-regulated in degenerative NP tissues and in IL-1β-stimulated NP cells, as compared to the levels in normal controls (normal tissue specimens from patients with idiopathic scoliosis). Bioinformatics and luciferase activity analyses showed that mothers against decapentaplegic homolog 3 (SMAD3), a key mediator of the transforming growth factor-β signaling pathway, was directly targeted by miR-15b...
December 30, 2016: Biotechnology Letters
Liangcai Lin, Yong Chen, Jingen Li, Shanshan Wang, Wenliang Sun, Chaoguang Tian
OBJECTIVES: To elucidate the mechanism of cellulase signal transduction in filamentous fungi including the components of the cellulase induction pathway. RESULTS: Neurospora crassa ncw-1 encodes a non-anchored cell wall protein. The absence of ncw-1 increased cellulase gene expression and this is not due to relieving carbon catabolite repression mediated by the cre-1 pathway. A mutant lacking genes encoding both three major β-glucosidase enzymes and NCW-1 (Δ3βGΔncw-1) was constructed...
December 30, 2016: Biotechnology Letters
Fabiany da Costa Gonçalves, Mateus Grings, Natália Schneider Nunes, Fernanda Otesbelgue Pinto, Tuane Nerissa Alves Garcez, Fernanda Visioli, Guilhian Leipnitz, Ana Helena Paz
OBJECTIVE: To investigate the effects of oxidative stress injury in dextran sulfate sodium (DSS)-induced colitis in mice treated with mesenchymal stem cells (MSC). RESULTS: Mice exposed to oral administration of 2% DSS over 7 days presented a high disease activity index and an intense colonic inflammation. Systemic infusion of MSC protected from severe colitis, reducing weight loss and diarrhea while lowering the infiltration of inflammatory cells. Moreover, toxic colitis injury increased oxidative stress...
December 28, 2016: Biotechnology Letters
Sam Mathew, Saravanan Prabhu Nadarajan, Uthayasuriya Sundaramoorthy, Hyunwoo Jeon, Taeowan Chung, Hyungdon Yun
OBJECTIVE: To enzymatically synthesize enantiomerically pure β-amino acids from β-keto nitriles using nitrilase and ω-transaminase. RESULTS: An enzyme cascade system was designed where in β-keto nitriles are initially hydrolyzed to β-keto acids using nitrilase from Bradyrhizobium japonicum and subsequently β-keto acids were converted to β-amino acids using ω-transaminases. Five different ω-transaminases were tested for this cascade reaction, To enhance the yields of β-amino acids, the concentrations of nitrilase and amino donor were optimized...
December 21, 2016: Biotechnology Letters
Lijuan Wang, Rihe Peng, Yongsheng Tian, Man Liu, Quanhong Yao
OBJECTIVES: To find an L-glutamate oxidase (LGox), to be used for the quantitative analysis of L-glutamic acid, an lgox gene encoding LGox from Streptomyces diastatochromogenes was isolated, cloned and characterized. RESULTS: The gene had an ORF of 1974 bp encoding a protein of 657 amino acid residues. In comparison to the LGox precursor, the proteinase K-treated enzyme exhibited improved affinity to substrate and with a K m of 0.15 mM and V max of 62 μmol min(-1) mg(-1)...
December 20, 2016: Biotechnology Letters
Tingzhen Mu, Maohua Yang, Jixiang Zhao, Moustafa Mohammed Sharshar, Jiangnan Tian, Jianmin Xing
OBJECTIVE: To construct efficient transformation and expression system and further improve desulfurizing activity of cells through expression of Vitreoscilla hemoglobin (VHb) in haloalkaliphilic Thialkalivibrio versutus SOB306. RESULTS: We transferred plasmids pKT230 and pBBR-sm(r) into T. versutus SOB306 via a conjugation method. We identified four promoters from among several predicted promoters by scoring for streptomycin resistance, and finally selected tac and p3 based on the efficiency of expression of red fluorescent protein (RFP)...
December 20, 2016: Biotechnology Letters
Yangyang Chen, Mengjie Liu, Shouwen Chen, Xuetuan Wei
OBJECTIVES: To reduce the unpleasant odor during 1-deoxynojirimycin (DNJ) production, the genes of leucine dehydrogenase (bcd) and phosphate butryltransferase (ptb) were deleted from Bacillus amyloliquefaciens HZ-12, and the concentrations of branched-chain short fatty acids (BCFAs) and DNJ were compared. RESULTS: By knockout of the ptb gene, 1.01 g BCFAs kg(-1) was produced from fermented soybean by HZ-12Δptb. This was a 56% decrease compared with that of HZ-12 (2...
December 20, 2016: Biotechnology Letters
Xianzhang Jiang, Hongjiao Liu, Yongchao Niu, Feng Qi, Mingliang Zhang, Jianzhong Huang
OBJECTIVES: To enlarge the diversity of the desaturases associated with PUFA biosynthesis and to better understand the transcriptional regulation of desaturases, a Δ(6)-desaturase gene (Md6) from Mucor sp. and its 5'-upstream sequence was functionally identified in Saccharomyces cerevisiae. RESULTS: Expression of the Δ(6)-fatty acid desaturase (Md6) in S. cerevisiae showed that Md6 could convert linolenic acid to γ-linolenic acid. Computational analysis of the promoter of Md6 suggested it contains several eukaryotic fundamental transcription regulatory elements...
December 17, 2016: Biotechnology Letters
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