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Biotechnology and Applied Biochemistry

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https://www.readbyqxmd.com/read/29679420/a-dual-label-time-resolved-fluorescence-immunoassay-for-the-simultaneous-determination-of-carcino-embryonic-antigen-and-squamous-cell-carcinoma-antigen
#1
Qian Mao, Xiaohong Liu, 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 carcino-embryonic 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
https://www.readbyqxmd.com/read/29664204/characterization-and-epitope-mapping-of-a-panel-of-monoclonal-antibodies-against-hiv-1-matrix-protein
#2
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
https://www.readbyqxmd.com/read/29644748/amidated-scolopin-2-inhibits-proliferation-and-induces-apoptosis-of-hela-cells-in-vitro-and-in-vivo
#3
Weili Yan, Jia Lu, Guiting Li, Huiyuan Wei, Wen-H Ua 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 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 (s.c.) 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
https://www.readbyqxmd.com/read/29633344/identification-and-characterization-of-a-novel-esterase-from-thauera-sp
#4
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 belong 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
https://www.readbyqxmd.com/read/29633336/production-and-characterization-of-a-human-lysosomal-recombinant-iduronate-2-sulfatase-produced-in-pichia-pastoris
#5
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-D Íaz, 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
https://www.readbyqxmd.com/read/29575133/gemcitabine-induces-apoptosis-and-autophagy-via-the-ampk-mtor-signaling-pathway-in-pancreatic-cancer-cells
#6
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
https://www.readbyqxmd.com/read/29572945/structural-basis-of-non-canonical-polyphenol-oxidase-activity-in-dll-ii-a-lectin-from-dolichos-lablab
#7
Yashavanth L V, 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. While 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
https://www.readbyqxmd.com/read/29569756/kinetic-models-for-the-effect-of-temperature-on-flavonoid-production-in-liquid-submerged-fermentation-by-phellinus-baumii
#8
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
https://www.readbyqxmd.com/read/29569743/the-psychrotrophic-yeast-sporobolomyces-roseus-lock-119-as-a-source-of-a-highly-active-aspartic-protease-for-the-in-vitro-production-of-antioxidant-peptides
#9
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 approx. 560 U/L. Optimization of medium composition and process temperature considerably enhanced enzyme biosynthesis; an approx. 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
https://www.readbyqxmd.com/read/29543347/development-and-characterization-of-a-camelid-single-domain-antibody-directed-to-human-cd22-biomarker
#10
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
https://www.readbyqxmd.com/read/29457659/improved-production-purification-and-characterization-of-biosurfactants-produced-by-serratia-marcescens-sm3-and-its-isogenic-smrg-5-strain
#11
Nashbly Sarela Rosas-Galván, Fernando Martínez-Morales, Silvia Marquina-Bahena, Raunel Tinoco-Valencia, Leobardo Serrano-Carreón, Brandt Bertrand, Renato León-Rodríguez, Josefina Guzmán-Aparicio, Laura Alvaréz-Berber, María R Trejo-Hernández
In this study, the biosurfactants (Bs) production of two Serratia marcescens strains (SM3 and its isogenic SMRG-5 strain) was improved and the tenso-active agents were purified and characterized. A 23 factorial design was used to evaluate the effect of nitrogen and carbon sources on the surface tension (ST) reduction and emulsion index (EI24 ) of the produced Bs. Optimum Bs production by SM3 was achieved at high concentrations of carbon and nitrogen, reducing ST to 26.5 ± 0.28 dynes/cm, with an EI24 of 79...
February 19, 2018: Biotechnology and Applied Biochemistry
https://www.readbyqxmd.com/read/29427529/efficient-bioconversion-of-oleuropein-from-olive-leaf-extract-to-antioxidant-hydroxytyrosol-by-enzymatic-hydrolysis-and-high-temperature-degradation
#12
Min Liu, Qiang Yong, Shiyuan Yu
Hydroxytyrosol (HT), a powerful antioxidant, clears free radicals and exhibits many biological activities. Because contents of HT are low in natural sources, bioconversion of oleuropein (OLE) to HT is of increasing interest. A biotechnological process was investigated to produce HT from OLE presented in olive leaf extract. Enzymatic hydrolysis using two cellulases with high β-glucosidase activity, Novozymes CTec2 and commercial cellulase KDN (Qingdao, People's Republic of China) was carried out at 50 °C for 12 H followed by raising the temperature to 90 °C for chemical hydrolysis...
February 10, 2018: Biotechnology and Applied Biochemistry
https://www.readbyqxmd.com/read/29405478/extracting-functional-groups-of-allini-to-design-derivatives-of-fda-approved-drugs-inhibition-of-hiv-1-integrase
#13
Umesh Kalathiya, Monikaben Padariya, Maciej Baginski
HIV-1 integrase (IN) is crucial for integration of viral DNA into the host genome and a promising target in development of antiretroviral inhibitors. In this work, six new compounds were designed by linking the structures of two different class of HIV-1 IN inhibitors (active site binders and allosteric IN inhibitors (ALLINIs)). Among newly designed compounds, INRAT10b was found most potent HIV-1 IN inhibitor considering different docking results. To further validate protein-ligand interactions obtained from dockings, molecular dynamics simulations were performed for inhibitor raltegravir and INRAT10b placed either at active site or allosteric site of HIV-1 IN (monomer or dimer)...
February 6, 2018: Biotechnology and Applied Biochemistry
https://www.readbyqxmd.com/read/29393538/process-optimization-of-the-extraction-condition-of-%C3%AE-amylase-from-brewer-s-malt-and-its-application-in-the-maltose-syrup-production
#14
Chengtuo Niu, Feiyun Zheng, Yongxian Li, Chunfeng Liu, Qi Li
β-Amylase is of important biotechnological aid in maltose syrup production. In this study, the extraction condition of β-amylase from brewer's malt and the optimal dosage of β-amylase in maltose syrup production were optimized using response surface methodology and uniform design method. The optimal extraction condition of β-amylase from brewer's malt was composed of 1:17 (g/v) material/liquid ratio, 44°C extraction temperature, pH 6.4 buffer pH, 2.3 H extraction time, and 1.64 g L-1 NaSO3 dosage with a predicted β-amylase activity of 1,290...
February 2, 2018: Biotechnology and Applied Biochemistry
https://www.readbyqxmd.com/read/29378087/advances-in-ginsenoside-biosynthesis-and-metabolic-regulation
#15
REVIEW
Jun Lu, Jinxin Li, Shihui Wang, Lu Yao, Wenxia Liang, Juan Wang, Wenyuan Gao
In this paper, we reviewed the advances in ginsenoside biosynthesis and metabolic regulation. To begin with, the application of elicitors in the ginsenoside biosynthesis was discussed. Methyl jasmonate (MJ) and analogues have the best effect on accumulation of ginsenoside compared with other elicitors, and few biotic elicitors are applied in Panax genus plants tissue culture. In addition, so far, more than 40 genes encoding ginsenoside biosynthesis related enzymes have been cloned and identified from Panax genus, such as UDP-glycosyltransferases (UGT) genes UDPG, UGTAE2, UGT94Q2, UGTPg100, and UGTPg1...
January 29, 2018: Biotechnology and Applied Biochemistry
https://www.readbyqxmd.com/read/29377386/effect-of-linkers-on-immobilization-of-scfvs-with-biotin-streptavidin-interaction
#16
Svetlana P Ikonomova, Megan T Le, Neha Kalla, Amy J Karlsson
Single-chain variable fragment antibodies (scFvs) are attractive for use in applications that require high specificity and binding to a target, such as biosensors. Previously, we demonstrated that a variety of scFvs can be immobilized onto a streptavidin surface through in vivo biotinylation of the biotin carboxyl carrier protein (BCCP) or smaller AviTag fused to the scFvs. However, the BCCP constructs showed better immobilization than the AviTag constructs. In this work, we investigated whether the discrepancy between the biotinylation tags could be alleviated by incorporating a flexible (G4 S)n linker of varying lengths or a rigid (EA3 K)3 linker between the biotinylation tags and the scFvs scFv13R4 and scFv5...
January 29, 2018: Biotechnology and Applied Biochemistry
https://www.readbyqxmd.com/read/29377374/identification-of-small-molecular-inhibitors-for-efflux-protein-rv2688c-of-mycobacterium-tuberculosis
#17
Rajender Vadija, Kiran Kumar Mustyala, Vasavi Malkhed, Ramasree Dulapalli, Hymavathi Veeravarapu, Ramesh Malikanti, Uma Vuruputuri
Mycobacterium tuberculosis (Mtb) is the pathogen, which causes tuberculosis. The development of multidrug-resistant and extensively drug-resistant strains in Mtb is due to an efflux mechanism of antibiotics in the bacteria. The efflux pump proteins in the bacteria are implicated in the active efflux of antibiotics. The efflux pump protein, "fluoroquinolones export ATP-binding protein Rv2688c" (FEAB), is considered as a potential therapeutic target to prevent tuberculosis. In the present work, in silico protocols are applied to identify inhibitors for the FEAB protein to arrest the efflux mechanism...
January 29, 2018: Biotechnology and Applied Biochemistry
https://www.readbyqxmd.com/read/29377329/efficient-production-and-characterization-of-homopolymeric-poly-3-hydroxyvalerate-produced-by-bacillus-strain-pjc48
#18
Jin-Jin Gu, Ying Zhou, Yan-Bin Tong, Jian-Jiang Lu, Bang-Ce Ye
Aliphatic polyester, poly(3-hydroxyvalerate) (PHV), is commonly produced as a granular component in bacterial cells of various species. Based on 16S rDNA gene sequence analysis, strain PJC48 was identified as a Bacillus species. The current study is aimed to screen for a high-yield strain that can produce PHV efficiently and to increase PHV product yield by optimizing the fermentative process. We identified a high-producer strain based on Nile red staining. Characterization of the PHV produced by PJC48 by nuclear magnetic resonance spectroscopy revealed that it consisted of (R)-3-hydroxyvalerate monomers...
January 29, 2018: Biotechnology and Applied Biochemistry
https://www.readbyqxmd.com/read/29357405/combat-biofilm-by-bacteriostatic-aptamer-functionalized-graphene-oxide
#19
Biyao Mao, Lijuan Cheng, Sheng Wang, Jiaqi Zhou, Le Deng
Biofilms are the main reason for a large number deaths and high health costs. Their better protection compared to planktonic form against conventional antibiotics leads to poor treatment efficiency. Nanoagent-targeted delivery is a promising avenue for disease therapeutic, but its application targeting biofilms has not been reported currently. The roles, if any, of aptamers acting as delivery carrier and targeting factor, the graphene oxide (GO), and GO modified with aptamers against biofilms were then systematically evaluated...
January 22, 2018: Biotechnology and Applied Biochemistry
https://www.readbyqxmd.com/read/29341247/secretory-expression-of-negative-regulatory-region-of-human-notch1-in-e-coli-and-preparation-of-a-functional-polyclonal-antibody
#20
Dengyang Zhang, Tianzhu Sun, Jingyun Yan, Xuanjun Wang, Jun Sheng
Notch signaling is a highly conserved pathway existed in multicellular organisms. It plays roles in normal human body development, human cancer initiation, progression and metastasis. The Notch negative regulatory region (NRR) is critical for Notch signaling, and cleavage at the S2 site in the NRR ultimately leads to the activation of Notch signaling. To study the function of human NRR1, we expressed the recombinant human NRR1 (rhNRR1) domain in Escherichia coli. After purification, rhNRR1 was obtained with approximately 94% purity according to SDS-PAGE analysis...
January 17, 2018: Biotechnology and Applied Biochemistry
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