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Biotechnology and Applied Biochemistry

Aidong Ruan, Yuan Gao, Chao Fang, Yaofei Xu
Nicotine is a potent parasympathomimetic stimulant and an important natural alkaloid mainly found in the Nicotiana genus of plants. It can directly threaten ecological security and human health in tobacco waste, waste water and other forms of tobacco production. Therefore, it is the basis of nicotine pollution prevention and of great application value to explore efficient and wide range of nicotinophilic bacteria for tobacco industry and environmental protection.In this study, one nicotinophilic bacteria was isolated from the soil which accumulated tobacco waste over 50 years at Hefei cigarette factory...
July 4, 2018: Biotechnology and Applied Biochemistry
Meghdad Abdollahpour-Alitappeh, Sepand Razavi-Vakhshourpour, Mohsen Abolhassani
Acute myeloid leukemia (AML) is a clonal hematologic malignancy arising from a small population of leukemic cells initiating the disease. CD123 is differentially expressed in AML blasts compared with normal hematopoietic stem and progenitor cells. The aim of this study was to develop specific monoclonal antibodies (mAbs) directed against AML. Three BALB/c mice were immunized with the human CD123 antigen, and the immune spleen cells were fused with the SP2/0 myeloma cell line. Hybridomas were screened by indirect ELISA, and the positive hybrids were cloned by limiting dilution...
July 4, 2018: Biotechnology and Applied Biochemistry
Anastasija Panic, Julijana Stanimirovic, Milan Obradovic, Emina Sudar-Milovanovic, Milan Perovic, Milena Lackovic, Nina Petrovic, Esma R Isenovic
This study aimed to investigate in vivo effects of estradiol on the regulation of hepatic inducible nitric oxide synthase (iNOS) expression in the high fat (HF) diet-induced obesity. Also, we aimed to investigate whether activation of the extracellular regulated kinase (ERK1/2), adenosine monophosphate-activated protein kinase (AMPK), Src kinase, and miR-221 is involved in estradiol-mediated regulation of iNOS in the liver of obese male Wistar rats. Male Wistar rats were fed a standard laboratory diet or a HF diet for ten weeks...
June 29, 2018: Biotechnology and Applied Biochemistry
Siti Nor Syairah Anis, Mohd Suffian Mohd Annuar, Khanom Simarani
Biosynthesis and in vivo depolymerization of intracellular medium-chain-length poly-3-hydroxyalkanoates (mcl-PHA) in Pseudomonas putida Bet001 grown on lauric acid were studied. Highest mcl-PHA fraction (>50 % of total biomass) and cell concentration (8 g L-1 ) were obtained at carbon-to-nitrogen (C/N) ratio 20, starting cell concentration 1 g L-1 , and 48 H fermentation. The mcl-PHA comprised of 3-hydroxyhexanoate (C6 ), 3-hydroxyoctanote (C8 ), 3-hydroxydecanoate (C10 ), and 3-hydroxydodecanoate (C12 ) monomers...
May 28, 2018: Biotechnology and Applied Biochemistry
Jelena Spasic, Mina Mandic, Jelena Radivojevic, Sanja Jeremic, Branka Vasiljevic, Jasmina Nikodinovic-Runic, Lidija Djokic
Biocatalytic potential of Streptomyces strains isolated from the rhizosphere of plants and from mycorrhizosphere of fungi has been investigated. A total of 118 Streptomyces isolates were selected and functionally screened for 10 different biotechnologically important enzymatic activities: hydrolase (cellulase, cutinase, gelatinase, lipase, protease, polyhydroxyalkanoate (PHA) depolymerase), phenol oxidase and peroxidase (laccase, tyrosinase, and lignin peroxidase), and aminotransferase. Out of 118 tested Streptomyces spp...
May 24, 2018: Biotechnology and Applied Biochemistry
Qian Mao, Xiaohong Liu, CuiCui Chen, Jingwen Ye, Huankun Liang, Binxian Li, Xinyi Sun, Lin Sun, Yuman Sun, Xiaomei Mu, Laiqing Li
Among women worldwide, cervical cancer is the second-most common cancer, and cervical smears and DNA detection have low sensitivity or are too expensive. The concentrations of carcinoembryonic antigen (CEA) and squamous cell carcinoma (SCC) in the serum were detected using a sandwich immunoassay. The CEA and SCC in the serum were captured by anti-CEA and anti-SCC antibodies. After combining other anti-CEA- and anti-SCC-labeled antibodies with europium (III) (Eu3+ ) and samarium (III) (Sm3+ ) chelates, CEA and SCC were detected with time-resolved fluorometry (TRF)...
April 20, 2018: Biotechnology and Applied Biochemistry
Zhiqing Zhang, Feng Zhang, Shimeng Bai, Jiaming Qiao, Honglin Shen, Fang Huang, Shuangquan Gao, Shaowei Li, Ying Gu, Ningshao Xia
The HIV-1 Gag precursor protein (p55) is the main structural protein comprising the matrix (MA/p17), capsid (CA/p24), and nucleocapsid (NC/p7) proteins, and is uniquely responsible for virion assembly within the virus life cycle. The MA protein plays a critical role in plasma membrane targeting and envelope glycoprotein (Env) uptake during virion assembly. Yet, when viral infection occurs, the MA protein may also be involved in virion uncoating, dissociating from the plasma membrane, and participating in the nuclear importation process...
April 17, 2018: Biotechnology and Applied Biochemistry
Weili Yan, Jia Lu, Guiting Li, Huiyuan Wei, Wen-Hua Ren
This study aimed to investigate the effect of Scolopin-2, a cationic antimicrobial peptide from centipede venoms, and amidated Scolopin-2 on Hela cell viability in vitro and in vivo. The cellular proliferation was investigated with the MTT assay. Confocal laser scanning, flow cytometry, and Western blot analysis were employed to localize Scolopin-2-NH2 in Hela cells and to study the caused cells apoptosis. We subcutaneously injected Hela cells into BALB/c nude mice and studied if Scolopin-2-NH2 suppressed tumor growth in the mice...
April 12, 2018: Biotechnology and Applied Biochemistry
Niu Yu, Jin-Chang Yang, Guang-Tian Yin, Rong-Sheng Li, Wen-Tao Zou, Chang He
A novel esterase gene TLip was identified from the strain Thauera sp. and expressed at high levels in Escherichia coli. The TLip protein shared the highest identity (48%) to esterase TesA from Pseudomonas aeruginosa when compared to enzymes with reported properties. Phylogenetic analysis showed that TLip belongs to the GDSL family of bacterial lipolytic enzymes. TLip was an alkaline esterase with a broad optimal temperature range 37-50 °C and an optimal pH of 8.0. Substrate specificity assays showed that TLip preferred medium chain p-nitrophenyl esters (C6 -C12 )...
April 6, 2018: Biotechnology and Applied Biochemistry
Natalia Pimentel, Alexander Rodríguez-Lopez, Sergio Díaz, Juan C Losada, Dennis J Díaz-Rincón, Carolina Cardona, Ángela J Espejo-Mojica, Aura M Ramírez, Fredy Ruiz, Patricia Landázuri, Raúl A Poutou-Piñales, Henry A Cordoba-Ruiz, Carlos J Alméciga-Díaz, Luis A Barrera-Avellaneda
Hunter syndrome (Mucopolysaccharidosis II, MPS II) is an X-linked lysosomal storage disease produced by the deficiency of the lysosomal enzyme iduronate-2-sulfatase (IDS). Currently, MPS II patients are mainly treated with enzyme replacement therapy (ERT) using recombinant enzymes produced in mammalian cells. As an alternative, several studies have shown the production of active and therapeutic forms of lysosomal proteins in microorganisms. In this paper, we report the production and characterization of a recombinant IDS produced in the yeast Pichia pastoris (prIDS)...
April 6, 2018: Biotechnology and Applied Biochemistry
Jinhui Zhu, Yan Chen, Yun Ji, Yuanquan Yu, Yun Jin, Xiaoxiao Zhang, Jiale Zhou
Gemcitabine (GEM)-based chemotherapy is a commonly used treatment for pancreatic cancer. However, acquired drug resistance, a major problem in pancreatic cancer treatment, causes a reduction in the survival rate of patients with cancer. In this study, we attempted to reveal the molecular mechanism of GEM resistance. Our data showed that GEM treatment inhibits cell growth, induces apoptosis, and activates autophagy via the AMP-activated protein kinase (AMPK) pathway. The combination of GEM treatment and AMPK knockdown resulted in a dramatic increase of apoptosis and inhibition of autophagy...
March 25, 2018: Biotechnology and Applied Biochemistry
Yashavanth L Vishweshwaraiah, Abhishek Acharya, Balaji Prakash
Lectins known to possess an additional enzymatic function are called leczymes. Previous studies reported a unique polyphenol oxidase (PPO) activity in DLL-II-a leczyme from Dolichos lablab. DLL-II shares a high sequence and structural homology with DBL-another leczyme from Dolichos biflorus. Incidentally, DBL possesses lipoxygenase activity, but not the PPO activity. Legume lectins usually possess two metal-binding sites A and B. Although these sites are conserved in both DBL and DLL-II, site A in DLL-II is occupied by Mn2+ and site B by Ca2+ ...
March 23, 2018: Biotechnology and Applied Biochemistry
Fu-Chun Jiang, He-Nan Zhang, Di Wu, Na Feng, Zhong Zhang, Jing-Song Zhang, Jie Feng, Yan Yang
Kinetic models and temperature control strategy were established to reflect the effect of temperature (22 °C-30 °C) on flavonoid production of Phellinus baumii (P. baumii) in 6-L fermentor. A modified Logistic equation, Hinshelwood model, and Luedeking Piret equation were used to describe mycelial growth and product formation. The influence of temperature on the estimated kinetic parameters was further studied by regression analysis. Based on kinetic parameters analysis, the new temperature control strategy was proposed...
March 23, 2018: Biotechnology and Applied Biochemistry
Aneta M Białkowska, Joanna Krysiak, Tomasz Florczak, Katarzyna M Szulczewska, Marta Wanarska, Marianna Turkiewicz
A psychrotrophic yeast strain producing a cold-adapted protease at low temperature was classified as Sporobolomyces roseus. In standard YPG medium, S. roseus LOCK 1119 synthesized an extracellular protease with an activity of approximately 560 U/L. Optimization of medium composition and process temperature considerably enhanced enzyme biosynthesis; an approximate 70% increase in activity (2060 U/L). The native enzyme was purified to homogeneity by cation exchange chromatography followed by a size exclusion step, resulting in a 103-fold increase in specific activity (660 U/mg) with 25% recovery...
March 23, 2018: Biotechnology and Applied Biochemistry
Fatemeh Faraji, Nader Tajik, Mahdi Behdani, Mohammad Ali Shokrgozar, Amir Hassan Zarnani, Fatemeh Shahhosseini, Mahdi Habibi-Anbouhi
CD22 is a B-cell-specific trans-membrane glycoprotein, which is found on the surface of the most B cells and modulates their function, survival, and apoptosis. Recently, targeting this cell surface biomarker in B-cell malignancies and disorders has attracted a lot of attention. The variable domain of camelid single-chain antibodies (VHH, nanobody) is a form of antibodies with novel properties including small size (15-17 kDa), thermal and chemical stability, high affinity and homology to human antibody sequences...
March 15, 2018: Biotechnology and Applied Biochemistry
Dongsheng Sun, Yang Liu, Hao Wu, Ying Ren, Xiaojun Ma, Huijian Wu, Guangwei Sun
Optimizing alginate gel beads is necessary to support the survival, proliferation, and function of entrapped hepatocytes. In this study, gelling bath was modified by decreasing calcium ion concentration and increasing sodium ion concentration. Alginate gel beads (using 36% G sodium alginate) prepared in the modified gelling bath had more homogeneous structure and better mass transfer properties compared with the traditional gelling bath that contains only calcium ions. Moreover, alginate gel beads generated in the modified gelling bath could significantly promote the HepG2 cell proliferation and the growth of cell spheroids, and maintain the albumin secretion ability similar to alginate gel beads prepared in the traditional gelling bath with only calcium ions...
March 2018: Biotechnology and Applied Biochemistry
Ning Wang, Long Wang, Lianwen Qi, Xu Lu
Triterpene saponins are main active constituents of Panax notoginseng. Metabolites profiling of 12 triterpene saponins was analyzed by high-performance liquid chromatography-mass spectrometry in leaf, petiole, and root extracts of P. notoginseng. Most of the 20(S)-protopanaxatriol (PPT) type saponins, except ginsenoside Re, were mainly distributed in roots, while 20(S)-protopanaxadiol (PPD) type saponins were detected among various tissues. The total content of PPD-type saponins decreased in the order of leaf, petiole, and root...
March 2018: Biotechnology and Applied Biochemistry
Maryam Khatami, Seyed Nezamedin Hosseini, Sadegh Hasannia
Alpha-1-antitrypsin (A1AT) is a major serum protein in human with protease inhibitory activity. Because of its extensive application in medicine, recombinant DNA technology has been considered for its production. The current study examines coexpression of A1AT and soluble domain of v-SNARE in Pichia pastoris, which can prevent the secretion of A1AT after thoroughly passing the secretory pathway. This was done mainly to preserve the biological activity of A1AT, which in the secretory mode might be impaired in the fermentation and early clarification conditions...
March 2018: Biotechnology and Applied Biochemistry
Bo Zhang, Hai-Dong Zhang, Yi-Teng Zhou, Kai Huang, Zhi-Qiang Liu, Yu-Guo Zheng
Amphotericin B is an effective antifungal antibiotic. However, its production in the wild-type strain of Streptomyces nodosus is relatively low. In this study, a strain of ZJB 20130827, capable of producing amphotericin B, was isolated and identified as S. nodosus based on its physiological and biochemical characteristics as well as its 16S rRNA gene sequence analysis. With N-methyl-N-nitroso-N'-nitroguanidine (NTG) and ultraviolet (UV) treatment, this wild-type strain was mutated for improving the yield of amphotericin B...
March 2018: Biotechnology and Applied Biochemistry
Parisa Farrokh, Bagher Yakhchali, Ali Asghar Karkhane
Thermostable lipases have many applications in detergent industries and in organic synthesis. There are many ways to improve thermal stability of enzymes, for example, higher hydrophobicity, greater structural packing, higher content of the charged residues, and lower thermolabile ones. In this study, thermolabile Gln (sensitive to higher temperatures) was substituted with Ala in native ELBn12 and mutated K173A lipases to examine its effect on thermal stability and activity of the lipases. Single (Q177A) and double mutants (K173A/Q177A) were expressed in Escherichia coli pLysS...
March 2018: Biotechnology and Applied Biochemistry
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