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Protein shake

Quan Li, Yuhang Zhang, Du Dechao, Yu Yanfei, Wei Zhang
Streptococcus suis, an important swine pathogen and a major zoonotic agent, is responsible for severe financial losses in the global swine industry. Although a multitude of virulence factors have been reported, the pathogenesis of S. suis infections remains poorly understood. In our previous work, we identified a potential virulence-associated protein, named PnuC, unique to virulent strains of S. suis serotype 2 (S. suis 2). To investigate the functions of PnuC, the pnuC gene deletion mutant (ΔpnuC) was constructed in S...
March 2018: Veterinary Microbiology
Wen Zhu, Guihua Gong, Jie Pan, Shu Han, Wei Zhang, Youjia Hu, Liping Xie
Human serum albumin (HSA) has been extensively used in a series of clinical care settings for nearly seven decades. However, the broad application of this protein is seriously limited by its short supply. In this work, the codon sequence of HSA was cloned under the control of the alcohol oxidase 1 promoter (AOX1) and expressed as a secretory protein in Pichia pastoris. A recombinant strain displaying the highest HSA yield was selected by screening for resistance to the highest concentration of antibiotic G418...
March 5, 2018: Protein Expression and Purification
Sarah L Irons, Adam C Chambers, Olga Lissina, Linda A King, Robert D Possee
Baculovirus expression systems are well established as an easy and reliable way to produce high quality recombinant proteins. Baculoviruses can also be used to transduce mammalian cells, termed 'BacMam', with considerable potential in biomedical applications. This chapter explains the process of making a recombinant baculovirus, encompassing production of a recombinant virus by homologous recombination in insect cells, followed by amplification and titration of the virus-all steps needed before commencing gene expression and protein production...
February 21, 2018: Current Protocols in Protein Science
Sean M McClure, Patrick L Ahl, Jeffrey T Blue
PURPOSE: The purpose was to evaluate DSF for high throughput screening of protein thermal stability (unfolding/ aggregation) across a wide range of formulations. Particular focus was exploring PROTEOSTAT® - a commercially available fluorescent rotor dye - for detection of aggregation in surfactant containing formulations. Commonly used hydrophobic dyes (e.g. SYPRO™ Orange) interact with surfactants, complicating DSF measurements. METHODS: CRM197 formulations were prepared and analyzed in standard 96-well plate rT-PCR system, using SYPRO™ Orange and PROTEOSTAT® dyes...
March 5, 2018: Pharmaceutical Research
Chao Fan, Xinxin Xu, Liya Song, Weishi Guan, Jinyang Li, Bo Liu, Pengjun Shi, Wei Zhang
A transfer DNA (T-DNA)-tagged mutant library of Humicola insolens was screened for mutants with altered cellulase production using the plate-clearing zone assay. Three selected mutants (5-A7, 5-C6, and 13-B7) exhibited significantly depressed FPase, CMCase and xylanase activities compared with the wild-type strain upon shake-flask fermentation, while the pNPCase and pNPGase activities of the three mutants were relatively higher than those of the parental strain. Combined with the results of SDS-PAGE and mass spectrometry, we suggest that expression of the CMCases Cel6B, Cel7B, CMC3, and XynA/B/C was reduced in the mutant strains...
March 2018: 3 Biotech
G Aravindhan, C Soundararajan, K Nagarajan, S Gomathinayagam
Excretory and secretory antigen of adult Cotylophoron cotylophorum and Gastrothylax crumenifer was prepared and characterized by SDS-PAGE. Adult paramphistomes were collected in PBS from rumen of sheep slaughtered at Perambur and Villivakkam slaughter houses, Chennai, Tamil Nadu. Excretory/Secretory (E/S) antigens of G. crumenifer and C. cotylophorum were prepared by incubating in PBS at 28.3 °C overnight in a shaking incubator and centrifuged. The culture supernatant was used as antigen. The protein concentration of E/S antigens of C...
March 2018: Journal of Parasitic Diseases: Official Organ of the Indian Society for Parasitology
Rachel Shirazi, Daniela Ram, Aviya Rakovsky, Efrat Bucris, Yael Gozlan, Yaniv Lustig, Pninit Shaked-Mishan, Orit Picard, Yonat Shemer-Avni, Haim Ben-Zvi, Ora Halutz, Yoav Lurie, Ella Veizman, Matthias Carlebach, Marius Braun, Michal Cohen- Naftaly, Amir Shlomai, Rifaat Safadi, Ella Mendelson, Ella H Sklan, Ziv Ben-Ari, Orna Mor
BACKGROUND: Characteristics of hepatitis B (HBV) and delta (HDV) coinfection in various geographical regions, including Israel, remain unclear. Here we studied HDV seroprevalence in Israel, assessed HDV/HBV viral loads, circulating genotypes and hepatitis delta antigen (HDAg) conservation. METHODS: Serological anti HDV IgG results from 8969 HBsAg positive individuals tested in 2010-2015 were retrospectively analyzed to determine HDV seroprevalence. In a cohort of HBV/HDV coinfected (n=58) and HBV monoinfected (n=27) patients, quantitative real-time PCR (qRT-PCR) and sequencing were performed to determine viral loads, genotypes and hepatitis delta antigen (HDAg) protein sequence...
February 27, 2018: BMC Infectious Diseases
Gerard J Marek, Brian P Ramos
5-Hydroxytryptamine2A (5-HT2A ) receptors are enriched in layers I and Va of the rat prefrontal cortex and neocortex and their activation increases the frequency of glutamatergic excitatory post-synaptic potentials/currents (EPSP/Cs) onto layer V pyramidal cells. A number of other G-protein coupled receptors (GPCRs) are also enriched in cortical layers I and Va and either induce (α1 -adrenergic and orexin2 ) or suppress (metabotropic glutamate2 [mGlu2 ], adenosine A1 , μ-opioid) both 5-HT-induced EPSCs and head twitches or head shakes induced by the phenethylamine hallucinogen 2,5-dimethoxy-4-iodoamphetamine (DOI)...
2018: Frontiers in Pharmacology
Hagar Mor-Shaked, Rachel Eiges
Fragile X syndrome (FXS) is one of the most common heritable forms of cognitive impairment. It results from a fragile X mental retardation protein (FMRP) protein deficiency caused by a CGG repeat expansion in the 5'-UTR of the X-linked FMR1 gene. Whereas in most individuals the number of CGGs is steady and ranges between 5 and 44 units, in patients it becomes extensively unstable and expands to a length exceeding 200 repeats (full mutation). Interestingly, this disease is exclusively transmitted by mothers who carry a premutation allele (55-200 CGG repeats)...
2018: Frontiers in Molecular Neuroscience
Tania Selas Castiñeiras, Steven G Williams, Antony Hitchcock, Jeffrey A Cole, Daniel C Smith, Tim W Overton
As high-level recombinant protein production (RPP) exerts a massive stress on the production host, an extensive literature on RPP optimization focuses on separating the growth phase from RPP production once sufficient biomass has been obtained. The aim of the current investigation was to optimize the benefits of the relatively neglected alternative strategy to achieve high-level RPP during growth by minimizing stress on the host. High yields of the biopharmaceutical recombinant human tumour necrosis factor alpha (rhTNFα) were obtained by fed-batch fermentation relevant to industrial production based upon parameters that most severely affected RPP in preliminary laboratory scale batch cultures...
February 14, 2018: Microbiology
Liangcheng Jiao, Qinghua Zhou, Zhixin Xu, Li Xu, Yunjun Yan
Rhizopus oryzae lipase (ROL) is an important industrial enzyme limited in application due to its low production in native strains. Here, we used a new combined strategy to overexpress ROL in Pichia pastoris. An efficient method based on bio-brick was developed to construct a series of vectors harboring different copy numbers of ROL gene cassettes, which were then transformed into P. pastoris GS115 to generate a strain with specific copy numbers of ROL. An optimized gene-dosage recombinant strain of GS115/pAOα-5ROL 11# harboring five copies of ROL was screened, revealing production of the highest activity (2700 U/mL), which was 8-fold higher than that of the strain harboring one copy...
February 13, 2018: Protein Expression and Purification
Zhiliang Yang, Zisheng Zhang
Background: 2,3-butanediol (2,3-BD) is a bulk platform chemical with various potential applications such as aviation fuel. 2,3-BD has three optical isomers: (2R, 3R)-, (2S, 3S)- and meso-2,3-BD. Optically pure 2,3-BD is a crucial precursor for the chiral synthesis and it can also be used as anti-freeze agent due to its low freezing point. 2,3-BD has been produced in both native and non-native hosts. Several pathogenic bacteria were reported to produce 2,3-BD in mixture of its optical isomers including Klebsiella pneumoniae and Klebsiella oxytoca ...
2018: Biotechnology for Biofuels
Joo Shun Tan, Sahar Abbasiliasi, Saeid Kadkhodaei, Yew Joon Tam, Teck-Kim Tang, Yee-Ying Lee, Arbakariya B Ariff
BACKGROUND: Demand for high-throughput bioprocessing has dramatically increased especially in the biopharmaceutical industry because the technologies are of vital importance to process optimization and media development. This can be efficiently boosted by using microtiter plate (MTP) cultivation setup embedded into an automated liquid-handling system. The objective of this study was to establish an automated microscale method for upstream and downstream bioprocessing of α-IFN2b production by recombinant Escherichia coli...
January 4, 2018: BMC Microbiology
Rajat Pandey, Venkata Dasu Veeranki
Biosimilar/Biotherapeutic production is becoming a major area of focus for a big chunk of Biotechnology Industry. Easy licensing and already approved status for clinical use has given it a boost. In the present study, recombinant human interferon gamma was expressed for the first time in Kluyveromyces Lactis (K. lactis) expression system and its expression was optimized by varying growth parameters and carbon source concentration with the aim of increasing recombinant protein production level. Human Interferon gamma gene (IFNG) was cloned in the genomic DNA of K...
February 9, 2018: Preparative Biochemistry & Biotechnology
Lisa A Spacek, Arthur Strzepka, Saurabh Saha, Jonathan Kotula, Jeffrey Gelb, Sarah Guilmain, Terence Risby, Steven F Solga
Ammonia physiology is important to numerous disease states including urea cycle disorders and hepatic encephalopathy. However, many unknowns persist regarding the ammonia response to common and potentially significant physiologic influences, such as food. Our aim was to evaluate the dynamic range of ammonia in response to an oral protein challenge in healthy participants. We measured blood and breath ammonia at baseline and every hour for 5.5 hours. Healthy men (N = 22, aged 18 to 24 years) consumed a 60 g protein shake (high dose); a subset of 10 consumed a 30 g protein shake (moderate dose) and 12 consumed an electrolyte drink containing 0 g protein (control)...
February 7, 2018: Scientific Reports
Kun He, Hong Sun, Hao Song
OBJECTIVES: To enhance the yield of 9α-hydroxy-4-androstene-3,17-dione (9-OHAD) from phytosterols, a phytosterol transport system was constructed in Mycobacterium sp. strain MS136. RESULTS: 9-OHAD can be produced via the controlled degradation of phytosterols by mycobacteria. This involves an active transport process that requires trans-membrane proteins and ATP. A phytosterol transport system from Mycobacterium tuberculosis H37Rv was constructed in Mycobacterium sp...
February 1, 2018: Biotechnology Letters
Gopalakrishnamurty Kadambari, Lakshminarayana R Vemireddy, Akkareddy Srividhya, Ranjithkumar Nagireddy, Siddhartha Swarup Jena, Mahendranath Gandikota, Santosh Patil, Roja Veeraghattapu, D A K Deborah, G Eswar Reddy, Maliha Shake, Aleena Dasari, P V Ramanarao, Ch V Durgarani, C N Neeraja, E A Siddiq, Maganti Sheshumadhav
A major dwarfing region for plant height, asd1, was identified employing the next-generation sequencing-based QTL-Seq approach from a dwarf mutant and is demonstrated to be responsible for the dwarf nature with least penalty on yield in rice. The yield plateauing of modern rice is witnessed since many decades due to the narrow genetic base owing to the usage of a single recessive gene, i.e., semi-dwarf-1 (sd-1) for development of short-statured varieties throughout the world. This calls for the searching of alternate sources for short stature in rice...
January 31, 2018: Plant Cell Reports
Hui Zhang, Xiao Sun, Wenjie Li, Tuoping Li, Suhong Li, Motomitsu Kitaoka
SPase is widely used in the food, cosmetics, and pharmaceutical industries. Previously, a SPase gene was cloned from Bifidobacterium longum JCM1217 and constructed into Escherichia coli BL21. In this paper, its expression conditions were optimized. The results showed that several induction factors determined the expression efficiency of SPase. The initial cell density, IPTG concentration, and induction time and temperature significantly (p < 0.01) affected the total protein content and activity of expressed SPase...
January 29, 2018: Protein Journal
Xi Wang, Ee-Been Goh, Harry R Beller
BACKGROUND: We previously developed an E. coli strain that overproduces medium-chain methyl ketones for potential use as diesel fuel blending agents or as flavors and fragrances. To date, the strain's performance has been optimized during growth with glucose. However, lignocellulosic biomass hydrolysates also contain a substantial portion of hemicellulose-derived xylose, which is typically the second most abundant sugar after glucose. Commercialization of the methyl ketone-producing technology would benefit from the increased efficiency resulting from simultaneous, rather than the native sequential (diauxic), utilization of glucose and xylose...
January 27, 2018: Microbial Cell Factories
Stephana Carelli, Mattia Colli, Valeriano Vinci, Fabio Caviggioli, Marco Klinger, Alfredo Gorio
The adipose tissue is a source of inflammatory proteins, such as TNF, IL-6, and CXCL8. Most of their production occurs in macrophages that act as scavengers of dying adipocytes. The application of an orbital mechanical force for 6-10 min at 97 g to the adipose tissue, lipoaspirated and treated according to Coleman procedures, abolishes the expression of TNF-α and stimulates the expression of the anti-inflammatory protein TNF-stimulated gene-6 (TSG-6). This protein had protective and anti-inflammatory effects when applied to animal models of rheumatic diseases...
January 16, 2018: International Journal of Molecular Sciences
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