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https://www.readbyqxmd.com/read/28894268/recycling-of-a-selectable-marker-with-a-self-excisable-plasmid-in-pichia-pastoris
#1
Cheng Li, Ying Lin, Xueyun Zheng, Qingyan Yuan, Nuo Pang, Xihao Liao, Yuanyuan Huang, Xinying Zhang, Shuli Liang
Pichia pastoris is a widely used heterologous protein production workhorse. However, with its multiple genetic modifications to solve bottlenecks for heterologous protein productivity, P. pastoris lacks selectable markers. Existing selectable marker recycling plasmids have drawbacks (e.g., slow growth and conditional lethality). Here, zeocin-resistance marker recycling vectors were constructed using the Cre/loxP recombination system. The vectors were used to (i) knock in heterologous phytase, xylanase and lipase expression cassettes, (ii) increase the phytase, xylanase and lipase gene copy number to 13, 5, and 5, respectively, with vector introduction and (iii) engineer the secretion pathway by co-overexpressing secretion helper factors (Sly1p and Sec1p) without introducing selectable markers, giving a phytase field of 0...
September 11, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28861598/screening-and-characterization-of-phytases-from-bacteria-isolated-from-chilean-hydrothermal-environments
#2
Milko A Jorquera, Stefanie Gabler, Nitza G Inostroza, Jacquelinne J Acuña, Marco A Campos, Daniel Menezes-Blackburn, Ralf Greiner
Phytases are enzymes involved in organic phosphorus cycling in nature and widely used as feed additives in animal diets. Thermal tolerance is a desired property of phytases. The objectives of this study were to screen and characterize bacterial phytases from Chilean hydrothermal environments. In this study, 60% (30 of 63) of screened thermophilic (60 °C) isolates showed phytase activity in crude protein extracts. The characterization of phytase from two selected isolates (9B and 15C) revealed that both isolates produce phytases with a pH optimum at 5...
August 31, 2017: Microbial Ecology
https://www.readbyqxmd.com/read/28858377/effects-of-%C3%AE-irradiation-on-cotyledon-cell-separation-and-pectin-solubilisation-in-hard-to-cook-cowpeas
#3
Talknice Z Jombo, Amanda Minnaar, John Rn Taylor
BACKGROUND: Cowpeas stored under high temperature and humidity develop the hard-to-cook defect (HTC). This defect greatly increases cooking times and energy costs. To better understand the mechanisms involved in the HTC defect development, the effects of γ-irradiation on cotyledon cellular structure and pectin solubility in two cowpea cultivars with different susceptibility to HTC defect were investigated. RESULTS: Gamma-irradiation decreased cotyledon cell wall thickness, increased cell size intercellular spaces in both cowpea cultivars and reduced cooking time of the less HTC susceptible cultivar...
August 31, 2017: Journal of the Science of Food and Agriculture
https://www.readbyqxmd.com/read/28854754/effects-of-multi-carbohydrase-and-phytase-on-standardized-ileal-digestibility-of-amino-acids-and-apparent-metabolizable-energy-in-canola-meal-fed-to-broiler-chicks
#4
C Gallardo, J C Dadalt, E Kiarie, M A Trindade Neto
Two assays were conducted to evaluate nutritive value of canola meal (CM) fed to broiler chicks without or with a multi-carbohydrase (MC) preparation (700 U α-galactosidase, 2,200 U galactomannanase, 30,000 U xylanase, and 22,000 U β-glucanase per kg of diet) and phytase (Phy, 500 FTU per kg of diet). Assay 1 determined apparent total tract digestibility (ATTD) of nutrients and metabolizable energy (AME) by the difference method. Assay 2 determined apparent ileal digestibility (AID) and standardized ileal digestibility (SID) of amino acids by the index method...
September 1, 2017: Poultry Science
https://www.readbyqxmd.com/read/28835691/proteomic-analysis-of-phytase-transgenic-and-non-transgenic-maize-seeds
#5
Yanhua Tan, Zheng Tong, Qian Yang, Yong Sun, Xiang Jin, Cunzhi Peng, Anping Guo, Xuchu Wang
Proteomics has become a powerful technique for investigating unintended effects in genetically modified crops. In this study, we performed a comparative proteomics of the seeds of phytase-transgenic (PT) and non-transgenic (NT) maize using 2-DE and iTRAQ techniques. A total of 148 differentially expressed proteins (DEPs), including 106 down-regulated and 42 up-regulated proteins in PT, were identified. Of these proteins, 32 were identified through 2-DE and 116 were generated by iTRAQ. It is noteworthy that only three proteins could be detected via both iTRAQ and 2-DE, and most of the identified DEPs were not newly produced proteins but proteins with altered abundance...
August 23, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28821305/establishment-of-a-decaplex-pcr-capillary-gel-electrophoresis-method-for-the-simultaneous-detection-of-six-kinds-of-genetically-modified-animals
#6
Xiaofei Liu, Songyin Qiu, Xiaolin Li, Dandan Liu, Hongli Jing, Qin Wang, Xiangmei Lin, Dengke Pan, Ningning Shi
This study aimed to establish an event-specific multiplex PCR system using microsatellite markers and fluorescently labeled primers to detect six different genetically modified (GM) animal lines, including human lactoferrin GM cattle, human lysozyme GM cattle, human α-lactalbumin GM cattle, myostatin knockout pigs, phytase GM pigs, and ω-3 fatty acid desaturase gene GM pigs. Four different microsatellite loci for species identification, along with six GM animal-specific fragments, were selected as targets for primer design...
August 18, 2017: Journal of AOAC International
https://www.readbyqxmd.com/read/28814823/relationship-between-the-antifungal-susceptibility-profile-and-the-production-of-virulence-related-hydrolytic-enzymes-in-brazilian-clinical-strains-of-candida-glabrata
#7
Maria Helena Galdino Figueiredo-Carvalho, Lívia de Souza Ramos, Leonardo Silva Barbedo, Jean Carlos Almeida de Oliveira, André Luis Souza Dos Santos, Rodrigo Almeida-Paes, Rosely Maria Zancopé-Oliveira
Candida glabrata is a facultative intracellular opportunistic fungal pathogen in human infections. Several virulence-associated attributes are involved in its pathogenesis, host-pathogen interactions, modulation of host immune defenses, and regulation of antifungal drug resistance. This study evaluated the in vitro antifungal susceptibility profile to five antifungal agents, the production of seven hydrolytic enzymes related to virulence, and the relationship between these phenotypes in 91 clinical strains of C...
2017: Mediators of Inflammation
https://www.readbyqxmd.com/read/28755320/evaluation-of-the-mature-grain-phytase-candidate-hvpaphy_a-gene-in-barley-hordeum-vulgare-l-using-crispr-cas9-and-talens
#8
Inger B Holme, Toni Wendt, Javier Gil-Humanes, Lise C Deleuran, Colby G Starker, Daniel F Voytas, Henrik Brinch-Pedersen
In the present study, we utilized TALEN- and CRISPR/Cas9-induced mutations to analyze the promoter of the barley phytase gene HvPAPhy_a. The purpose of the study was dual, validation of the PAPhy_a enzyme as the main contributor of the mature grain phytase activity (MGPA), as well as validating the importance of a specific promoter region of the PAPhy_a gene which contains three overlapping cis-acting regulatory elements (GCN4, Skn1 and the RY-element) known to be involved in gene expression during grain filling...
July 28, 2017: Plant Molecular Biology
https://www.readbyqxmd.com/read/28752758/engineering-of-yersinia-phytases-to-improve-pepsin-and-trypsin-resistance-and-thermostability-and-application-potential-in-the-food-and-feed-industry
#9
Canfang Niu, Peilong Yang, Huiying Luo, Huoqing Huang, Yaru Wang, Bin Yao
Susceptibility to proteases usually limits the application of phytase. We sought to improve the pepsin and trypsin resistance of YeAPPA from Yersinia enterocolitica and YkAPPA from Y. kristensenii by optimizing amino acid polarity and charge. The predicted pepsin/trypsin cleavage sites F89/K226 in pepsin/trypsin-sensitive YeAPPA and the corresponding sites (F89/E226) in pepsin-sensitive but trypsin-resistant YkAPPA were substituted with S and H, respectively. Six variants were produced in Pichia pastoris for catalytic and biochemical characterization...
August 30, 2017: Journal of Agricultural and Food Chemistry
https://www.readbyqxmd.com/read/28748480/investigation-of-the-effects-of-starch-on-the-physical-and-biological-properties-of-polyacrylamide-paam-starch-nanofibers
#10
Hadi Taghavian, Seyed-Omid Ranaei-Siadat, Mohammad Reza Kalaee, Saeedeh Mazinani
Here, we report the development of a new polyacrylamide (PAAm)/starch nanofibers' blend system and highlight its potential as substrate for efficient enzyme immobilization. PAAm was synthesized and blended with starch. The final blend was then electrospun into nanofibers. The response surface methodology was used to analyze the parameters that control nanofiber's diameter. Electrospun mat was then modified either by cross-linking or phytase immobilization using silane coupling agent and glutaraldehyde chemistry...
July 26, 2017: Progress in biomaterials
https://www.readbyqxmd.com/read/28741666/assessment-of-the-potential-of-the-multi-enzyme-producer-bacillus-amyloliquefaciens-us573-as-alternative-feed-additive
#11
Ameny Farhat-Khemakhem, Monia Blibech, Ines Boukhris, Mohamed Makni, Hichem Chouayekh
BACKGROUND: Recently, probiotics have increasingly been used as feed additives in poultry diets as an alternative to antibiotic growth promoters fostering resistance development. RESULTS: This study was aimed at assessing the potential of Bacillus amyloliquefaciens US573 as a direct-fed microbial. The US573 strain was found to be free of harmful enzymatic activities and sensitive to antibiotics. In addition, it showed a good acid and bovine bile tolerance, high adhesion efficacy to chicken enterocytes, and an ability to form biofilms, which may favor its survival and persistence in the animal gastrointestinal tract...
July 25, 2017: Journal of the Science of Food and Agriculture
https://www.readbyqxmd.com/read/28727029/effects-of-graded-levels-of-microbial-phytase-on-apparent-total-tract-digestibility-of-calcium-and-phosphorus-and-standardized-total-tract-digestibility-of-phosphorus-in-four-sources-of-canola-meal-and-in-soybean-meal-fed-to-growing-pigs
#12
Y She, Y Liu, H H Stein
One hundred twenty pigs were used to determine effects of graded levels of microbial phytase on the apparent total tract digestibility (ATTD) of P and Ca and the standardized total tract digestibility (STTD) of P in 4 sources of canola meal and in 1 source of soybean meal (SBM) fed to growing pigs. The 4 sources of canola meal were produced from 1 source of high-protein canola seeds and 2 sources of conventional canola seeds with 1 of the conventional canola seeds being divided into 2 separate batches before crushing...
May 2017: Journal of Animal Science
https://www.readbyqxmd.com/read/28720951/isolation-and-characterization-of-lactic-acid-bacteria-from-traditional-pickles-of-himachal-pradesh-india
#13
Monika, Savitri, Vijay Kumar, Anila Kumari, Kunzes Angmo, Tek Chand Bhalla
A total of 15 isolates of lactic acid bacteria (LAB) were isolated from traditional pickles of rural and urban areas of Himachal Pradesh. These isolates were identified as Enterococcus faecalis (7), Lactobacillus plantarum (3), Pediococcus pentosaceus (2), Leuconostoc mesenteroides (1), Lactococcus lactis (1) and Enterococcus sp. (1) on the basis of morphological, biochemical and 16S rDNA sequence analysis. All the LABs produced exopolysaccharides, have antibiotic susceptibility, and possess activity of β-galactosidase, β-glucosidase, protease and amylase, while none of the isolates showed haemolytic activity...
June 2017: Journal of Food Science and Technology
https://www.readbyqxmd.com/read/28710591/effect-of-dietary-phytase-supplementation-on-bone-and-hyaline-cartilage-development-of-broilers-fed-with-organically-complexed-copper-in-a-cu-deficient-diet
#14
Siemowit Muszyński, Ewa Tomaszewska, Małgorzata Kwiecień, Piotr Dobrowolski, Agnieszka Tomczyk
Tibial mechanical, chemical, and histomorphometrical traits were investigated for growing male Ross 308 broiler chickens fed diets that had copper (Cu) from organic source at a lowered level of 25% of the daily requirement (4 mg kg(-1) of a premix) with or without phytase. Dietary treatments were control non-copper, non-phytase group (0 Suppl); 4 mg kg(-1) Cu non-phytase group (25%Cu); and 4 mg kg(-1) Cu + 500 FTU kg(-1) phytase group (25%Cu + phyt). The results show that birds fed with the addition of phytase exhibited improved weight gain and final body weight and had increased serum IGF-1 and osteocalcin concentrations...
July 15, 2017: Biological Trace Element Research
https://www.readbyqxmd.com/read/28669072/free-and-immobilized-aspergillus-oryzae-sbs50-producing-protease-resistant-and-thermostable-phytase
#15
Sapna, Bijender Singh
Optimization for enhanced phytase production by Aspergillus oryzae SBS50 in submerged fermentation was investigated using Taguchi design. In first step design, starch, beef extract, magnesium sulphate, ferrous sulphate and Tween 80 were identified as significant factors affecting phytase production. These significant factors were further optimized at four different levels using a second Taguchi design and were observed that 1% starch, 2% beef extact, 3% Tween 80, 0.1% magnesium sulphate and 0.225% ferrous sulphate supported maximum phytase production (47,432 U/L)...
July 2017: 3 Biotech
https://www.readbyqxmd.com/read/28667635/genetically-modified-phytase-crops-role-in-sustainable-plant-and-animal-nutrition-and-ecological-development-a-review
#16
REVIEW
Chinreddy Subramanyam Reddy, Seong-Cheol Kim, Tanushri Kaul
Globally, plant-derivatives especially cereals and legumes are the major staple food sources for animals. The seeds of these crops comprise of phytic acid, the major repository form of the phosphorus, which is not digestible by simple-stomached animals. However, it is the most important factor responsible for impeding the absorption of minerals by plants that eventually results in less use of fertilizers that ultimately cause eutrophication in water bodies. Although abundant phosphorus (P) exists in the soils, plants cannot absorb most of the P due to its conversion to unavailable forms...
July 2017: 3 Biotech
https://www.readbyqxmd.com/read/28667571/extracellular-secretion-of-phytase-from-transgenic-wheat-roots-allows-utilization-of-phytate-for-enhanced-phosphorus-uptake
#17
Samreen Mohsin, Asma Maqbool, Mehwish Ashraf, Kauser Abdulla Malik
A significant portion of organic phosphorus comprises of phytates which are not available to wheat for uptake. Hence for enabling wheat to utilize organic phosphorus in form of phytate, transgenic wheat expressing phytase from Aspergillus japonicus under barley root-specific promoter was developed. Transgenic events were initially screened via selection media containing BASTA, followed by PCR and BASTA leaf paint assay after hardening. Out of 138 successfully regenerated To events, only 12 had complete constructs and thus further analyzed...
August 2017: Molecular Biotechnology
https://www.readbyqxmd.com/read/28664910/corrigendum-engineering-the-residual-side-chains-of-hap-phytases-to-improve-their-pepsin-resistance-and-catalytic-efficiency
#18
Canfang Niu, Peilong Yang, Huiying Luo, Huoqing Huang, Yaru Wang, Bin Yao
This corrects the article DOI: 10.1038/srep42133.
June 30, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28660848/effects-of-rapeseed-meal-fiber-content-on-phosphorus-and-calcium-digestibility-in-growing-pigs-fed-diets-without-or-with-microbial-phytase
#19
M Bournazel, M Lessire, M J Duclos, M Magnin, N Même, C Peyronnet, E Recoules, A Quinsac, E Labussière, A Narcy
The optimization of dietary phosphorus (P) and calcium (Ca) supply requires a better understanding of the effect of dietary fiber content of co-products on the digestive utilization of minerals. This study was designed to evaluate the effects of dietary fiber content from 00-rapeseed meal (RSM) on P and Ca digestibility throughout the gastrointestinal tract in growing pigs fed diets without or with microbial phytase. In total, 48 castrated male pigs (initial BW=36.1±0.4 kg) were housed in metabolic crates for 29 days...
June 29, 2017: Animal: An International Journal of Animal Bioscience
https://www.readbyqxmd.com/read/28647122/effect-of-succinate-on-phosphate-solubilization-in-nitrogen-fixing-bacteria-harbouring-chick-pea-and-their-effect-on-plant-growth
#20
Bhagya Iyer, Mahendrapal Singh Rajput, Shalini Rajkumar
Diverse nitrogen fixing bacteria harbouring chick pea rhizosphere and root nodules were tested for multiple plant growth promoting traits like tricalcium phosphate (TCP) and rock phosphate (RP) solubilization, production of ammonia, indole 3-acetic acid, chitinase, phytase and alkaline phosphatase. Isolates belonged to diverse genus like Enterobacter, Acinetobacter, Erwinia, Pseudomonas, Rhizobium, Sinorhizobium, Ensifer, Klebsiella, etc. Most isolates solubilized TCP and RP along with the lowering of media pH, indicating acidification to be the chief mechanism behind this solubilization...
September 2017: Microbiological Research
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